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Full text of "Annual report"

ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October^'lS 1987 - SepCember 30, 1988 

REPORT OF THE SCIENTIFIC DIRECTOR 
Jin H. Kinoshica, Ph.D. 



During chis past year, we were very pleased that three of our NEI 
intramural scientists received well deserved recognition. The research 
accomplishments of these three scientists have brought great distinction to 
the NEI. 

Dr. Robert Wurtz, Chief of the Laboratory of Sensorimotor Research was 
elected to membership to the highly prestigious National Academy of Science. 
Dr. Wurtz 's outstanding contributions involve a series of experiments each of 
which made a pioneering advance in its area. He pioneered the use of 
conscious monkeys in the study of the visual and oculomotor systems. Years 
ago, he developed a technique which allows the visual system to be studied in 
conscious behaving animals and this technique is now a standard technique used 
throughout the world. Using this technique, Dr. Wurtz was the first to record 
single cells in the visual cortex of the awake monkey and thus to confirm in 
awake behaving monkeys the organization of cells in the anesthetized, 
paralyzed animals seen by Hubel and Wiesel to whom the Nobel Prize was given a 
few years ago. He went on to analyze the effect of eye movements on visual 
processing by single cells in the visual pathway. Dr. Wurtz was the first to 
study the mid-brain structure, the superior colliculus, which is one of the 
major destinations of the neurons leaving the retina of the eye, and he first 
determined that the cells in this structure were involved in both vision and 
eye movements. Dr. Wurtz was also the first to explore the relationship of 
the basal ganglia to the initiation of the eye movements. 

Throughout this work. Dr. Wurtz has combined behavioral analysis, 
physiological techniques, and histological controls to carry out the most 
sophisticated experiments possible. He and his associates introduced the use 
of on-line computers in the analysis of physiological function in a trained 
behaving animal. In all these experiments Dr. Wurtz has emphasized the 
functional approach to the nervous system, that is, how brain cells are 
organized to produce behavior. This in turn has allowed the ready application 
of the discoveries in the basic research laboratory to deficits seen in man in 
the clinic. It is this aspect that allows his laboratory within the National 
Eye Institute to be so closely associated with the clinic and to hold such 
promise for scientific breakthroughs directly relevant to the understanding of 
disease. His methodological contributions to the electrophysiological study 
of vision and oculomotor functions in awake, behaving monkeys are world 
renowned. 

Dr. Chader, Chief of the Laboratory of Retinal Cell and Molecular Biology, 
won the 1988 Friedenwald Award given by the Association for Research in Vision 
and Ophthalmology. Dr. Chader won acclaim for studies on two important 
hereditary diseases of the retina, retinitis pigmentosa and retinoblastoma. 

Retinitis pigmentosa is a hereditary blinding disease that selectively 
strikes the photoreceptor cells of the retina. Dr. Chader 's work has linked 



I 



abnormalities in cGMP metabolism to retinal degeneration in several animal 
models of the human disease. In particular, he pinpointed a deficit in cGMP- 
PDE activity in retinal photoreceptors in the early postnatal period well 
before the morphological signs of the disease become apparent. This seems to 
be of general significance since such deficits have now been observed in four 
RP animal models. Although suitable human retinal RP tissue has not yet been 
available, this work forms the basis for studies on the human disease and also 
as a model for other possible diseases in which there is abnormal cyclic 
nucleotide metabolism. 

Dr. Chader has focused on a novel protein that his laboratory first 
described in the retina in 1976, and has now been named the Interphotoreceptor 
Retinoid-Binding Protein, (IRBP). This retinoid-binding protein appears to be 
found only in the eye; it is synthesized by the retina but is quickly secreted 
into the subretinal space between the retinal photoreceptors and the adjacent 
pigment epithelial (PE) cell layer. Since vitamin A is stored in PE cell but 
is utilized in the photoreceptor cell for visual process, it is probable that 
this protein functions as an extracellular vehicle for retinoid transport 
between the two tissues. In fact, IRBP has many of the characteristics one 
would expect of such a transport vehicle including differential retinoid- 
binding in light and in dark. The protein having been isolated and fully 
characterized has been cloned in his laboratory. Dr. Chader and associates 
demonstrated that IRBP is capable of causing experimental autoimmune uveitis 
(EAU), a feature previously undescribed. Monkeys were found highly 
susceptible to IRBP-induced EAU. This monkey disease is of special interest 
because of the close pathological similarity to certain ocular diseases in 
man, in particular sympathetic ophthalmia and Vogt-Koyanagi-Harada disease. 
In addition to providing a useful model for the human diseases, the findings 
with IRBP-induced EAU in monkeys support the notion that autoimmune processes 
to retinal antigens participate in the etiology of certain human eye diseases. 

Laboratory of Mechanisms of Ocular Diseases 

Dr. J. Samuel Zigler, Jr., Head of the Cataract Research Section, has 
received a $50,000 award from the Alcon Research Institute. 

Dr. Zigler 's pioneering work has been on the mechanisms that account for 
the oxidative damage occurring in lens undergoing cataract formation. In the 
c'ase of senile nuclear cataracts, which are characterized by extensive 
oxidation of crystallins in the lens nucleus. Dr. Zigler was the first to show 
the possible role of singlet oxygen which is generated photodynamically within 
the lens. Exposure of crystallins to singlet oxygen produced oxidation of 
cysteine and tryptophan residues, the formation of non-disulf ide covalent 
crosslinks, the generation of an unusual non-tryptophan fluorescence, 
increased pigmentation, and aggregation of the proteins. AIL of these 
modifications closely resemble changes observed in crystallins from aging and 
cataractous human lenses. 

Cortical cataracts occur in the outer portion of the lens and may result 
from damage to cell membranes leading to osmotic swelling with consequent 
cataract formation. Dr. Zigler has investigated the roles of activated states 
of oxygen in such processes. Using a lens organ culture technique, he has 
investigated lens membrane damage produced by various "oxygen radical" 
generating systems. Exposure of lenses in vitro to concentrations of H2O2 
higher than those to which the lens is normally exposed in vivo, leads to 



impaired ability of the lenses to maintain normal cation balance which results 
in osmotic swelling and loss of transparency. 

The situation is quite different when activated species of oxygen are 
generated within the lens cells rather than in the surrounding fluids. Using 
solutions of lens proteins as a model of the intracellular environment. Dr. 
Zigler found that H2O2 alone produced little or no structural modifications to 
the crystallins. On the other hand when conditions were imposed to promote 
conversion of H0O2 into hydroxyl free radical, the crystallins were found to 
be rapidly modified. The modifications included covalent crosslink, formation, 
increased non-tryptophan fluorescence, aggregation, and changes in the net 
charge of the polypeptides. Thus the capacity for damage from the various 
oxygen radicals depends upon the environment. Within the cell the highly 
reactive hydroxyl free radical is extremely toxic since it is produced in the 
immediate vicinity of numerous target molecules. When generated outside the 
lens the stable species, H2O2, is most damaging because it can diffuse across 
ceil membranes. After entering the lens fibers the H2O2 likely interacts with 
metal ions, perhaps at specific metal binding sites on proteins, to generate 
hydroxyl free radical or related species which produce the actual protein 
damage. 

Although the research activities of these three scientists are undoubtedly 

outstanding, there are other research studies ongoing in an intramural program 

equally as exciting and important as one will glean by perusing this year's 
Annual Report. 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT ZOl EY 00065-11 OSD 



PERIOD COVERED 

October 1, 1987, to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must lit on one line between tne baraers.) 
Physiological studies of the Primate Visual System 



PRINCIPAL INVESTIGATOR (Ust other professional personnel Oe/ow me Pnncii^l Investigator.) (Name, title, latxratory, ana institute atfiliation) 
PI: Francisco M. de Monasterio, M.D., D.Sc. Medical Officer OSD, NEI 

Others: 



COOPERATING UNITS (it any) 



LAB/BRANCH 

Office of the Scientific Director 



SECTION 



INSTITUTE AND LOCATION 



NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

0.55 



PROFESSIONAL; 

0.55 



OTHER: 

0.00 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects D (b) Human tissues D (c) Neither 

n (a1) Minors 
n (a2) Interviews 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the apace provided.) 

The project involves the study of the physiological organization of neurons of 
the visual system of primates. Studies were carried out to characterize: (1) 
the chromatic organization of the peripheral region of the "center-surround" 
organization of the receptive field of color-opponent ganglion cells, and (2) 
the degree of heterogeneity of properties among color-opponent ganglion cells 
whose receptive fields are located in the retinal periphery. During the period 
covered, analyses of some prior studies were completed, and results are being 
prepared for publication. 

(1) Comparison of the results of area-threshold measurements and of chromatic 
mapping of the receptive field with a small test spot showed that a fraction 
of the color-opponent ganglion cells of macaque retina has antagonistic center 
and surround responses mediated in part by the same type of cone mechanism. 
The apparent frequency of these cells increases towards the retinal periphery, 
and their resulting center-surround organization provides a simple and direct 
model for the development of the recently reported "modified Type II" neurons 
of the striate cortex of macaques. 

(2) Further studies of the degree of homogeneity of peripheral color-opponent 
ganglion cells are consistent with the existence of two main cell classes that 
differ in terms of conduction velocity, receptive-field center size, and degree 
of surround antagonism. Preliminary results suggest that some cells of one of 
these groups loose color-opponent properties in the far periphery, developing 

a chromatic organization similar to that of the color non-opponent, broad-band 
ganglion cells. 



PHS 6040 (Rev. 1/84) GPO »I4-91« 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE ] 

NOTICE OF INTRAMURAL RESEARCH PROJECT i ZOI EY 00122-08 OSD 



PERIOD COVERED 

October 1, 1987, to September 30, 1988 



TITLE OF PROJECT (80 cnamcters or less. Title must tit on one line Detv/een tne txraers.) 

Anatomical Studies of the Primate Visual System 



PRINCIPAL INVESTIGATOR (Ust otner protessional personnel Below the Principal Investigator.) {Name, title, laboratory, ana institute attiliaoon) 

PI: Francisco M. de Monasterio, M.D., D.Sc. Medical Officer OSD, NET 
Other: 



COOPERATING UNITS (if any) 



Department of Ophthalmology, Georgetown University, DC (JC Horton, LR Dagi) 
Department of Ophthalmology, University of Washington, Seattle (A Bunt-Mylans) 



LAB/BRANCH 

Office of the Scientific Director 



SECTION 



INSTITUTE AND LOCATION 



NET, NTH, Bethesda. Maryland 20892 



TOTAL MAN-YEARS: 

0.45 



PROFESSIONAL: 

0.45 



OTHER: 
0.0 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects H (b) Human tissues D (c) Neither 

n (al) Minors 
n (a2) Interviews 



I 



SUMMARY OF WORK (Use stantiara unreducaO type. Do not exceed tfte space provided.) 

This project involves the study of the anatomical properties and organization 
of cells in the visual system of primates, with emphasis on the retina and the 
visual cortex. Studies were carried out to characterize (1) the eye-dominance 
column pattern of human visual striate cortex, (2) the correlation between the 
staining of blue-sensitive cones by anti-blue cones antibodies and by tissue- 
reactive dyes, and (3) the reported variability of cone density in the foveal 
region. 

(1) Despite the fact that the striate cortex of humans and macaques differ not 
only in terms of surface area, but also sulcal and gyral topography, cytochrome 
oxidase staining shows that the layout of the eye-dominance columns of striate 
cortex in patients who suffered monocular eye loss before death is very similar 
to that of macaques. These results indicate that the above anatomical factors 
do not determine the general pattern of eye-dominance columns. 

(2) Preliminary results of the comparison of the staining of a cone population 
by tissue-reactive dyes, and the labeling of blue-sensitive cones by anti-blue 
cone antibodies suggest that the putative identification of this population as 
blue-sensitive cones is indeed correct. 

(3) Results from cone density measurements in the fovea and area centralis of 
the retina of macaque and donor human eyes fail to substantiate, so far, major 
individual differences in cone density that have been claimed in some studies. 



PHS 6040 (Rev. 1/84) 



SPO 9l4-»lt 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT ZOl EY 00135-16 



PERIOD COVERED 

OcGOber 1. 1957 '2 Se"Dxember }2. 1938 



TITLE OF PROJECT (80 cftaracrers or less. Title must tit on one line IMtween the boraers.l 

rlocnemis'ry of Hexina and Pignenxed Zpixheliujii in Health and Disease 

PRINCIPAL INVESTIGATOR /List other protessionai personnel Delow tne Pnncipal Investigator) (Name, title, laboratory, ana institute affiliation) 

?I: Helen H. Hess :.I.D. Medical Officer (Research) OSD, NET 



COOPERATING UNITS (if any) 

Veterinary Resources Branch, DRS, NIH 



LAB/BRANCH 

Office of the Scientific Director, NEI 



SECTION 



INSTrrUTE AND LOCATION 

National Eye Institute, NIH, Bethesda, I.kryland 20892 



TOTAL MAN-YEARS; 

1.3 



PROFESSIONAL: 

1.0 



OTHER: 

0.3 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects D (b) Human tissues (c) Neither 

n (al) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Use stanaarO unreOucM type. Do not axcaed the space provided.) 

The effects of nutrition, oxidation, and other environmental factors (light 
intensity or darkness ) on the incidence and progress of posterior subcapsular 
opacities (PSO) associated with retinal degeneration are being studied in Royal 
College of Surgeons (RCS) rats, in which rod photoreceptor outer segment debris 
accumulates secondary to a phagocytic defect in the retinal pigmented epithelium. 
Evidence has been obtained that oxidative changes in polyunsaturated fatty acids 
in the debris lead to water-soluble toxic aldehydes that can be detected in the 
vitreous, and are toxic to lens cells and their membranes. Pink-eyed RCS 
dystrophic rats fed a natural ingredient diet (NIH-07) are highly sensitive to 
retinal light damage,, beginning at light levels as low as 1-4 footcandles, and 
27% of such rats develop mature cataracts by 7-12 months. Increased intensity of 
light ( either cyclic or constant ) increased the percentage of rats with mature 
cataracts, while dark rearing from birth prevented the PSO and mature cataracts. 
Recently, we have found that a purified diet (AIN-76A) fortified with O.A% beta- 
carotene + 0.01% BHT also prevented the PSO and mature cataracts. Rhosopsin 
bleaching appears to be essential for retinal light damage and for initiation of 
the PSO. A hypothesis has been developed that would explain these findings. It 
depends upon the known capacity of retinaldehyde to act as a photosensitizer to 
generate singlet oxygen, an extremely energetic oxidant for polyunsaturated 
lipids, as well as proteins. Darkness would prevent release of retinaldehyde, 
I while beta-carotene is a direct physical quencher of singlet oxygen, and BHT a 
■ highly efficient scavenger for secondary oxidized products. Principles 
established with the RCS rat model may have significance for slowing or prevent- 
ing human PSO and mature cataracts, such as those seen in retinitis pigmentosa. 



PHS 6040 (Rev 1/84) CPO 9i4.sit 



ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October 1, 1987 - September 30, 1988 

REPORT OF THE CLINICAL DIRECTOR 
Robert B. Nussenblatt, M.D. 



The Clinical Branch consists of two Sections, each with its own Section 
Head: Section on Ophthalmic Genetics and Pediatric Ophthalmology, Muriel I. 
Kaiser-Kupfer, M.D. and the Section on Retinal and Vitreal Diseases, Robert B. 
Nussenblatt, M.D. (Acting). 

The Section on Ophthalmic Genetics and Pediatric Ophthalmology was active 
in a wide range of activities. One area of major interest was the anterior 
segment. The short and long-term effects of contact lens wear on the cornea 
is actively being investigated. The changes in corneal curvature in corneal 
epithelium morphology as well as endothelial cell morphologry that may be 
induced with long term contact lens wear has great import for many individuals 
who use this method for correction of vision. Additionally, the group has 
developed objective and subjective methods to monitor and document opacities 
in the human lens using different systems. Reproducibility studies on 
objective systems include the use of the Scheimpflug cameras, the 
Retroillumination camera. Specular microscope and the laser light-scattering 
spectroscope. Other systems such as ultrastenography and nuclear magnetic 
residents (imaging) are being actively tested. The group is finding that it 
will be necessary to combine subjective and objective methods to characterize 
adequately the presence, progression or regression of cataracts. Many of the 
subjective methods that show promise include contrast sensitivity, potential 
acuity, glare, as well as a well-done visual acuity test. 

The group has been extremely interested in the posterior segment as well. 
The molecular genetics of retinal degenerations has been an area of particular 
interest. The intent is to identify the genes responsible for different 
inherited retinal disorders in animal models with the attempt to establish the 
genetic relationship of these animal disorders to forms of hiiman retinal 
degenerations . Work has centered on the rd and rds mutations in the mouse and 
the Abyssinian cat. The hope will be once the molecular basis of one or more 
of these animal models have inherited and retinal degenerations have been 
established, that this information will be applied to the human situation. 
With that in mind, the group has actively participated in the inter-institute 
medical genetics program and the genetics clinic. During the last year, 
approximately 400 individuals were seen representing approximately 100 
different disease categories. Because of the high frequency of ocular 
involvement in many of these cases, almost all of the patients were evaluated 
by the ophthalmic genetics staff or were discussed in consultation. The 
assessment of the posterior segment degenerative disorders is of upmost 
important and the group has actively pursued this goal. Objective 
measurements using electrophysiology techniques has demonstrated a wide 
variety of observations. Of note is the value of electroretinography in the 
early diagnosis of progressive cone dystrophy which was studied in a large 



number of three generations of a pedigree with dominant progressive cone 
dystrophy. The use of extensive testing has demonstrated that even at an 
early age in subjects from families with this disorder, while phychophysical 
and ophthalmoscopic criteria were insufficient to determine whether they were 
affected or not, the cone mediated ERG was clearly abnormal. It would 
certainly appear that until a genetic screening method becomes available the 
ERG is the earliest indicator of the presence of a cone dystrophy. Studies in 
gyrate atrophy of the choroid and retina continue. The continued accumulation 
of natural history data as well as the definition of the genetic abnormalities 
of this disorder provides us with continued important information in this 
area. A double-masked controlled randomized clinical trial of topical 
cysteamine has enrolled 16 patients. These individuals have been enrolled to 
test the efficacy of topical cysteamine (0.1% in humans) in order to see 
whether this will prevent the ocular manifestations of this disorder. Most 
specifically, the collection of crystals in the cornea. Four patients have 
shown a significant decrease in the cysteamine treated eyes and are now taking 
drops in both eyes. Recent work has demonstrated that the concentration of 
cysteamine could be increased to 0.5% with the results of this new dosage 
still awaited. 

The Section on Retinal Diseases and Vitreous remained heavily involved with 
two long clinical trials. The use of oral sorbinil, an aldose reductase 
inhibitor, continued to be tested in a randomized masked trial to see if it 
will inhibit diabetic retinopathy. This study was conducted simultaneously in 
ten research centers in the United States. Recruitment into this study has 
stopped and the results of the study will be awaited with great interest. 
Additionally, patients with macular degeneration continued to be studied in a 
randomized masked fashion in order to test the efficacy of vitamin E and C 
therapy as well as the prevention of damage from light below 500 nanometers in 
preventing this degenerative process. This is the leading cause of newly 
registered blindness in the white adult population in the United Sates. The 
recruited patients are examined at four month intervals with a follow-up to 
continue for five years unless an early beneficial or detrimental effect 
causes the study to be terminated in less than that time. Testing includes 
sterio fundus photographs of each macula once a year with the endpoint for the 
study that of visual acuity of 20/100 or less in the initially better eye 
because of disc form or atrophic degeneration of the macula. This study will 
continue until the needed number of patients have been recruited. 
Additionally, the section has been involved in the study of diabetic patients 
using vitreous f luorophotometry. Those without retinopathy, those with 
nonproliferative retinopathy, and normal volunteers have been studied. A new 
method for evaluating blood retinal barrier permeability to fluorescein and 
the diffusivity of fluorescein into the vitreous has been developed. 

The Clinical Branch reflects new horizons with basic research observations 
playing an increasingly greater role in the research being conducted. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



ZOl EY 00162-06 CB 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



, TITLE OF PROJECT ISO characters or less. Title must tit on one line Between tne Ixraers.l 



Vitreous irluorophotometry 



PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator) (Name, title, laboratory, ana institute atfiliation) 

PI: Monique S. Roy M.D. Visiting Scientist CB, NEl 



COOPERATING UNITS (H any) 

Peter Bungay Ph.D. BEIB, NIH 



U\B/BRANCH 

Clinical Branch 



SECTION 

Section on Retinal and Vitreal Diseases 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

0.4 



PROFESSIONAL; 

0.4 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

Q (a) Human subjects D (b) Human tissues D (c) Neither 

n (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Use standard unreduced type. Do not sxcaatf the apace provided.) 

Vitreous f luorophotometry has been performed in patients with diabetes 
mellitus without retinopathy, patients with diabetes mellitus with 
nonproliferative retinopathy, and normal volunteer subjects, age- and 
sex-matched to the patients. A new method for evaluating blood retinal 
barrier permeability to fluorescein and diffusivity of fluorescein in the 
vitreous has been developed. The amount of fluorescein leakage into the 
vitreous of patients has been compared to that of the normal subjects. 
Correlations with other features of diabetes, such as the quality of 
diabetic control, the existence of subclinical neuropathy and nephropathy, 
and other complications were sought. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



= OjECT \UMBEa 



ZOl EY 00198-05 CB 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must lit on one line Oetween the bonders.) 

Sorbinil Retinopathy Trial 



PRINCIPAL INVESTIGATOR (Ust otner professional personnel below the Principal Investigator) (Name, title, laboraton/. and institute atfiliation) 

PI: Monique S. Roy M.D. Visiting Scientist CB, NEI 



Others: Manuel Datiles M.D. 
James R. Carl M.D. 



Staff Ophthalmologist 
Senior Staff Fellow 



CB, NEI 
CB, NEI 



COOPERATING UNITS (H any) 

R. Silverman 



NIDDK, NIH 



UVB/BRANCH 

Clinical Branch 



SECTION 

Section on Retinal and Vitreal Diseases 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

0.7 



PROFESSIONAL: 

0.7 



CHECK APPROPRIATE BOX(ES) 

El (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



OTHER: 



D (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the apace pmyided.} 



Oral sorbinil, an aldose reductase inhibitor, will be administered in a 
double-masked randomized trial to diabetics with no or minimal diabetic 
retinopathy. This will be done to evaluate the effects of sorbinil on the 
development of diabetic retinopathy and further investigate the safety and 
toleration of sorbinil. The study will be conducted simultaneously in 10 
research centers in the USA. 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 

. Z01 gY 00187-05-C3 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 cnaracrers or less. Titie must tit on one line Between me ooraers.i 

The Effects of Corneal Contact Lenses on the Cornea 



i PRINCIPAL INVESTIGATOR lUsi oxner oroiessionai oersonnei oeiow tne Prmaoai invesiiaaior.) (Name, title, laooraiory. ana institute arliliationi 

I 



! PI: Manuel 3. Datiles M.D. Visiting Scientist 



CB, NSI 



j Others: Lessie McCain R.N. Clinical Technician CB NEI \ 

I Kayoko Kashima M.D. Visiting Associate CB,' NEI '[ 

! i 

! I 



COOPERATING UNITS (it any) 



\ LAB/BRANCH 

j Clinical Branch 



Section on Ophthalmic Genetics and Pediatric Ophthalmology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: i PROFESSIONAL: 

•15 I 0.10 



OTHER; 
.05 



CHECK APPROPRIATE BOX(ES) 

□ (a) Human subiects _ (b) Human tissues Z! (c) Neither 

LJ (a1) Minors 
G (a2) Interviews 

SUMMARY OF WORK CL/se stanaara unreaucea rype. Do not exceea tne space proviaeO.) 



Short- as well as long-term effects of contact lens wear on the cornea are being 
investigated. Changes in corneal curvature, changes in corneal eoithelial 
morphology and changes in corneal endothelial cell morphology are' being studied 
by specular microscopy. 

These data will help us understand the dynamics involved in the interaction 
between a contact lens and the cornea, the risk involved to corneal tissues, and 
how a systemic or local disorder may increase these risks. 



PHS 6040 (Rev 1/841 GPO»i«-9ie 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

Z01 EY 00138-05 CB 



i PERIOD COVERED 

! October 1, 1987 to September 30, 1988 



i TITLE OF PROJECT (80 cnaracters or less. Title musi tit on one line oetween tne ooraers.l 

i Documentation and Monitoring of Opacities in the Human Lens 

I PRINCIPAL INVESTIGATOR lUsi orner ororessionai oersonnei oeiow tne Pnnaoai investigator. ) f/Vame. title, /aooratory. ana institute arfiliationi 



?I: Manuel B. Datlles M.D. 

Others: Robert Sperduto M.D. 

Peter Kador Ph.D. 

Lessie McCain R.N. 



Visiting Scientist CB, NEI 

Head, Epidemiology Branch BEP, NEI 

Head, Section on LMOD, NEI 

Molecular Pharmacology 

Clinical Technician CB, NEI 



I COOPERATING UNITS III any) 

'Image Processing and Analysis Laboratory, DCRT, NIH (Benes Trus, Ph.D., Chief) 
] Clinical and Diagnostic Trials Section, NCI, NIH (Sylvan Green, M.D.) 
Nuclear Medicine, Clinical Center, NIH (Joseph Frank, M.D.) 



j u;b/branch 


' Clinical Branch 


i SECTION 


Section on Ophthalmic Genetics and Pediatric Ophthalmology 


INSTITUTE AND LOCATION 


NEI, NIH, Bethesda, Maryland 20892 


TOTAL MAN-YEARS; 


PROFESSIONAL: 


OTHER: 


1.2 


1.2 





CHECK APPROPRIATE BOX(ES) 

G (a) Human subjects [j (b) Human tissues D (c) Neither 

U (a1) Minors 
■H (a2) Interviews 



I SUMMARY OF WORK (Use stanaara unreaucea type. Do not axceeO tne space proviaea.) 



We are developing objective and subjective methods to monitor and document 
opacities in the human lens using different systems. We are presently actively 
recruiting patients with and without cataracts for reproducibility studies on 
the objective systems — the Scheimpflug cameras (Zeiss and topcon), Retrollluml- 
natlon camera (Neitz), Specular microscope (Keeler) and laser light-scattering 
spectroscope (KOWA). We will also test other systems using sound (ultrasono- 
gr^aphy), and nuclear magnetic resonance (magnetic resonance imaging). We are 
also studying subjective systems or methods, such as the effects of cataracts on 
visual perception, contrast sensitivity, and glare, which may be useful as 
additional parameters in the monitoring of cataract presence, progression, or 
regression. 



PHS 6040 (Rev 1/84) 



GPO 9 I 4-9 IS 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



'ZQI ;Y 00212-0^ C3 



; PERIOD COVERED 

'October 1, 1987 to September jO, 1988 



■ TITLE OF PROJECT ISO cnaracters or less. Tine musi til on one line oerween me ooraers.) 

Model --ogram for Collaboracion Between Cataract Surgeons and Ophthalmic Researcheri s 

j PRINCIPAL INVESTIGATOR lUsi orner oroiessionai oersonnei oeiow tne Pnncioai investigator. i iName. title, /aoorarofv. ana institute ariiliationi 

I 
1 

IpI: Manuel 3. Datiles M.D. Visiting Scientist CB, 



Others: Carl Kupfer M.D, 

Muriel I. Kaiser-Kuofer M.D, 



Director 

Head, Section on CB, 

Ophthalmic Genetics and 
Pediatric Ophthalmology 



NEI 

NEI 
NEI 



COOPERATING UNITS (it any) 

Jin H. Kinoshita 

W. Gerald Robison, Jr. 



Ph.D, 
Ph.D, 



Scientific Director 
Head, Section on 
Pathophysiology 



LMOD, 



NEI 
NEI 



j L^B/BRANCH 

'Clinical Branch 



; SECTION 




i Section on Ophthalmic Genetics and Pediatric 


Ophthalmology 


INSTITUTE AND LOCATION 




NEI, NIH, Bethesda, Maryland 20892 




TOTAL MAN-YEARS: 


PROFESSIONAL: 




OTHER: 


0.85 


0.85 






CHECK APPROPRIATE BOX(ES) 




n (a) Human subjects D (b) Human tissues 


□ (c) Neither 


C (a1) Minors 




n (a2) Interviews 





SUMMARY OF WORK lUse stanaara unreaucea type. Do not exceed tne space proviaeO.) 



There is presently an extreme scarcity of human cataract material because of an 
[abrupt shift of cataract surgical technique from intracapsular (intact lens) 
i to extracapsular (fragmented lens), primarily because of advent of the use of 
intraocular lens. We are exploring ways by which fragmented lens materials 
can be maximally used in cataract basic research through close collaboration 
with cataract surgeons and basic researchers and modification of techniques 
by both groups. 



PHS 6040 (Rev 1/841 



GPO SI4>9IS 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 EY 00246-01 CB 



PERIOD COVERED 

October i, 1987 to September 30, 1988 



TITLE OF PROJECT (80 cneraciers or less Title must In on one line Between tne ooraers i 

Molecular Genetics of Retinal Degenerations 



i PRINCIPAL INVESTIGATOR ILisl orner oroiessionei personnel Deiow tne Prmcioai investigator i IName tilie laooraiory ana institute atiiiiationi 

I 
I 



Michael 3. Gorin M.D., Ph.D. Medical Officer CB, NEI 

CB, NEI 



' ■!■: 
I 
I Others: Ignacio Rodriguez Ph.D. Staff Fellow 



I COOPERATING UNITS li< any/ 

Northwestern University (Larry Pinto, Ph.D.), University of Linkoping. Linkoping 
Chief? LVC;1ce! Nc'i)''"'^ ''''°"' '"''' Institute, (Stephen O'BrL, Ph.S"^' 



LAB/BRANCH 

Clinical Branch 


SECTION 

Section on Ophthalmic Genetics and Pediatric 


Ophthalmology 


INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 


TOTAL MAN-YEARS: 


PROFESSIONAL: 

1.9 


OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects G (b) Human tissues Zj (c) Neither 

D (a1) Minors 
[j (a2) Interviews 



SUMMARY OF WORK (Use stanaera unreaucea type. Do not exceeO tne space proviaea.) 



The purpose of this project is to identify the genes responsible for different 
inherited retinal disorders in animal models and to establish the genetic 
relationship of these animal disorders to forms ofThuman retinal degenerations. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



'01 SY ooon-ia cs 



PERIOD COVERED 

October 1, 1987 to Sept'ember 30, 1988 



TITLE OF PROJECT 180 cnaraciers or less- Title must tit on one line oerween me ooraers.i 

P i gmenL gispersion With and Without Glaucoma 



PRINCIPAL INVESTIGATOR (Usi otner oroiessionai personnel oeiow tne Prmaoai investigator i (Name, title, laoaraiory. ana institute artniationi 



I 



PI: 



Muriel I. Kaiser-Kupf er '^D. 



Others: Carl Kupfer 

Lessie McCain 



M.D. 
R.N. 



Head, Section on Ophthalmic 
Genetics and Pediatric 
Ophthalmology- 
Director 
Clinical Technician 



I 



:3, NEi 



NEI 
CB, NEI 



COOPERATING UNITS fil any) 



i LAB/BRANCH 



Clinical Brancn 



; SECTION 










Section on Oohthalm 


ic Genetic 


s and Pediatric 


0Dhthalmo1ns;v 


INSTITUTE AND LOCATION 










NEI, NIH, Bethesda, 


Maryland 


20892 






TOTAL MAN-YEARS; 


PROFESSIONAL: 




OTHER. 


0.25 




.15 




.? 



CHECK APPROPRIATE BOX(ES) 

G (a) Human subjects 
iZl (a1) Minors 
G {a2) Interviews 



(b) Human tissues 



G (c) Neither 



I SUMMARY OF WORK (Use sianaara unreaucea type. Do not exceea tne soace proviaea.i 



The purpose of this project is to determine the risks of patient with pigment 
dispersion syndrome to develping glaucoma. Comparisons of patients with and 
without glaucoma will be mde based on diagnostic tests, genetic screening, 
aqueous humor dynamics and pupillary responses to light. The data acquired 
may enable a determination of the risk of patients with pigment dispersion 
syndrome to developing glaucoma as well as add to the understanding of the 
pathology of the disease. 



PMS 6040 (Rev 1/841 



GPO 01 4-01 B 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



P^'OJECT NUMBER 



:01 EY 00062-12 CE 



PERIOD COVERED 

October 1 , 



987 to Seotember 



1988 



I TITLE OF PROJECT (30 cnaraciers or less. iitie musi in on one line oerween tne Donxrs.i 

i IriQo-Corneal-Endothelial (ICE) Syndrome 



PRINCIPAL INVESTIGATOR /Ust orner ororesstonai oersonnei oeiow me Prmaoai investigator! (Name, title, laooraton/. ana institute atliiiationi 



PI; 



Others: 



Muriel I. Kaiser-Kuof er M.D. 



Carl Kupfer 
Lessie McCain 
Manuel Datiles 



M.D 

R.N. 

M.D. 



Head, Section on Ophthalmic 
Genetics and Pediatric 
Ophthalmology 

Director 

Clinical Technician 

Visiting Scientist 



CB, NEI 



NEI 
CB, NEI 
CB. NEI 



COOPERATING UNITS (il any} 



LAB/BRANCH 

Clinical Branch 



SECTION 

Section on Ophthalmic Genetics and Pediatric Ophthalmology 



I INSTITUTE AND LOCATION 

i NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS; 
.25 



PROFESSIONAL; 



15 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects 
G (a1) Minors 
G (a2) Interviews 



D (b) Human tissues G (c) Neither 



SUMMARY OF WORK (Use stanaara unreaucea type. Do not exceaa tne soace proviaea.l 

This project was formerly titled "Progressive Essential Iris Atrophy." 
Patients are being recruited with progressive essential iris atrophy with or 
without associated corneal disease. Information is being gathered to evaluate 
the clinical features and course of the disease process and to investigate 
aqueous humor dynamics in both affected and unaffected eyes. 



PHS 6040 (Rev l/Sd) 



GPO 91 4-«18 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



Z01 EY 00083-1 1 CB 



PERIOD COVERED 

October 1, 1987 to September 30, 19? 



TITLE OF PROJECT (30 cnaracmrs or less. Title must ni on one line oerween tne ooraers.i 

Gyrate Atropny of the Choroid and Retina and Other Retinal Degenerations 

PRINCIPAL INVESTIGATOR (List orner ororessionai oersonnei oeiow me Pnncioai investigator.) (Name, title. laDoratory, ana institute attiliationt 

=1: Muriel I. Kaiser-Kupf er M.D. Head, Section on Ophthalmic 03, NEI 

Genetics and Pediatric 
Ophthalmology 

Others: Michael Gorln M.D., Ph.D. Medical Officer 

Lessie McCain R.-N. Clinical Technician 

Rafael Caruso M.D. Visiting Scientist 

Doris Collie A. A. Health Technician 



C3, NEI 

CB, NEI 

CB, NEI 

CB. NEI 



I COOPERATING UNITS (il any/ 

I The Howard Hughes Medical Institute Laboratory and the Department of Pediatrics 
j Johns Hopkins University, School of Medicine, Baltimore, Maryland 
' (David L. Valle, M.D.) 

! LAB/BRANCH 

, Clinical Branch 



SECTION 

Section on Ophthalmic Genetics and Pediatric Ophthalmology 

INSTITUTE AND LOCATION 

NEI, NIK, Bethesda, Maryland 20892 



TOTAL MAN- YEARS; 
1.2 



PROFESSIONAL; 
0.7 



OTHER; 
0.5 



CHECK APPROPRIATE BOX(ES) 

U (a) Human subjects Cj (b) Human tissues G (c) Neither 

G (a1) Minors 
G (a2) interviews 



SUMMARY OF WORK (Use stanaara unreaucea type. Do not exceed tne space proviaea.i 



Patients with gyrate atrophy of the choroid and retina are examined systematical- 
ly to confirm the diagnosis. Skin fibroblats of affected patients and family 
{ members are grown in tissue culture and assayed for ornithine aminotransferase 
■ activity. The results will be evaluated for correlation with the presence of 
homo- or heterozygosity for the disease trait. Patients will be given a trial 
of pyridoxine to see if serum concentration of ornithine can be reduced, and, if 
so, the patient will be classified as a "responder," and treatment with pyrido- 
xine will be continued. Nonresponder and responder patients will be placed on a 
low arginine, low protein, diet with supplemental amino acids and observed for an 
arrest or improvement of their disease. If patients are not considered eligible 
for the diet or if they appear unable to comply with the dietary regimen they will 
be followed to record the natural progress of the condition. Patients with other 
forms of retinal degeneration, such as retinitis pigmentosa, fundus flavimacula- 
tus, juvenile retinoschisis, are also examined and their courses are compared with 
gyrate atrophy patients. 



PHS 6040 (Rev 1/841 SPOSi4.»ia 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



'ZOl EX 00163-06 CB 



! PERIOD COVERED 

I October 1, 1987 to September ^0. 1^38 



! TITLE OF PROJECT tSO cnaracters or less Jnie must In on one line oerween me cxraers I 

i 

' NIH Interinstitute Medical Genetics Program: '^'le Gener.jps r.linin 

I PRINCIPAL INVESTIGATOR (US! omer oro'essionai oersonnei Deiow tne Pnnaoai Invesiigaior I (Name liiie. leooretory ana insmuie atliiiauoni 



Muriel I, Kalser-Kupf er M.D. 



Head, Section on Ophthalmic 03, NEI 



1 others: Michael B. 
i Lessie McC 


Go 
ain 


Genetics and Pediatric 
Ophthalmology 

rin M.D., Ph.D. Medical Officer 

R.N. Clinical Technician 


CB, 
CB, 


1 

NEI ; 

NEI 


1 COOPERATING UNITS (il any) 


i LAB/BRANCH 

i Clinical Brancn 


! SECTION 

Section on Ophthalm 


Lc Genetics and Pediatric Oohthalmoloev 






1 INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, 


Maryland 20892 






TOTAL MAN- YEARS: 
.2 


PROFESSIONAL; 
.1 


OTHER: 

.1 


CHECK APPROPRIATE BOXfES) 

D (a) Human subjects 
l] (a1) Minors 
D (a2) Interviews 


Q (b) Human tissues G (c) Neither 






1 SUMMARY OF WORK (Use stanaara unrea 
™. ^ . . .... ... 


ucea type. Do noi exceea tne soace proviaei 


'■) 







The Interinstitute Medical Genetics Program and the Genetics Clinic, supported 
by the Clinical Center, offer a multidisciplinary approach to patients with 
genetic disease (ZOl CP 05139-04 CEB). Involved in the program are researchers 
from all Institutes. Patients evaluated in the clinic represent a broad spectrum 
of .genetic diseases. During the last year, approximately 400 individuals were 
seen, representing approximately 100 different disease categories. Due to the 
high frequency of ocular Involvement in many of the cases, almost all the patients 
were evaluated by Clinical Branch staff or were discussed in consultation. The 
Clinic serves as a source of interesting case material concerning patients with 
inherited or developmental abnormalities of the visual system. 

In addition to the Genetics Clinic, patients are seen for genetic consultation 
at the Maryland School for the Blind. This experience has resulted the recruit- 
ment of patients into Clinical Branch protocols. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



»«Oject ,\oMBe= 



:01 Fx 00172-06 C3 



: PERIOD COVERED 

I October i, ^dZl to September JO, 1^: 



I TITLE OF PROJECT (80 cnarmemn or ««. Tioa muii m or on« on* o«iw»»n me oorvmrs.) 

I Acre Related Macular Deaeneration 



PRINCIPAL INVESTIGATOR (Uil otnmr oramMMionMi pertonnti omiow m* fnncioai Invsogator.) tNtrrm. arm. laoormtory, ana insmun attiuanoni 

PI: Muriel I. Kaiser-Kupfer M.D. Mead, Section on CB,l'd 

OohtiialLU-C Genetics 



Others: Carl Kuprer 

>tonique S. ?oy 



M.D. Director 

M.D. Visiting Scientist 



CB,NEI 




LAB/BRANCH 

Clinical Branch 



SECTION 

Section on Onhthalmic Genetics and Pediatric OoiitlialmDlocr/ 



INSTITUTE AND LOCATION 

NIH, riEI, Bethesda, :iarvland 20S92 



TOTAL MAN-YEARS: 
0.2 



PROFESSIONAL: 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

Q^ (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Un ttanatm unnaucae (yp«. Do not azcMfl me <oaca ptvmaae.) 

This Study will determine if patients with severe visual loss because of 
age related macular degeneration in one eye and with good vision in the second 
eye can be protected from severe visual loss in the good eye by the 
administration of vitamin E and vitamin C when exposure of the retina to light 
below 500 nanometers is diminished. The recruited patients will be randomly 
assigned either to a treated or untreated control group and examined at 
four-month intervals. Follow-up will continue for five years, unless an early 
beneficial or detrimental effect causes the study to be terminated in less than 
five years . 



PHS 6040 (Rev 1/84) 



PROJECT NUMBER 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



:oi EY 00123-08-CB 



PERIOD COVERED 

October 1, ;987 to September 30, 1988 



TITLE OF PROJECT {80 cnaraciers or less. iitie musi In on one line oerween me ooraers.) 

Clinical Psychophysics of the Visual System 



PRINCIPAL INVESTIGATOR lUst otner oroiessionai oersonnei oeiow tne Prinaoai invesiigaior.i (Name, iirie. laooratorv. ana insuiuie artiiiaKoni 



PI: 



I Others; 



Muriel I. Kaiser-Kupf er M.D. 



Rafael C. Caruso 
Kent E. Higgins 
Ralph D. Gunkel 



M.D. 

Ph.D. 

O.D. 



Head, Section on 
Ophthalmic Genetics 
and Pediatric Ophthalmology 

Visiting Scientist 

Expert 

Ophthalmic Physicist 



CB, NEI 



CB, NEI 
CB, NEI j 
CB, NEI 



COOPERATING UNITS (it anyi 

Georgetown University Center for Sight, Washington, D.C. (Despina Koutsandreas , 
O.T., Robert Toma, C.O.T.) 



LAB/BRANCH 

Clinical Branch 



SECTION 

Section on Ophthalmic Genetics and Pediatric Ophthalmology 



I INSTITUTE AND LOCATION 

I NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

1.3 



PROFESSIONAL. 

1.3 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
H (al) Minors 
~ {a2) Interviews 



n (b) Human tissues 



(c) Neither 



SUMMARY OF WORK (Use stanaara unreaucea type. Do not exceea tne soace proviaea.) 



The visual function of patients with ocular diseases or lesions in the visual 
pathways and of normal subjects is measured with psychophysical techniques. 
These data are correlated with those obtained with electrophysiological tests 
of visual function. The results obtained contribute to the diagnosis of ocular 
and neural disorders that affect vision, and are needed to characterize their 
nature and evolution. They are also valuable in the assessment of the effect of 
treatment regimens on the outcome of these diseases. 



PHS 6(M0 (Rev 1/841 



GPO 91 4.910 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 001i44-07-CB 



PERIOD COVERED 

October 1 , V 



]7 to September 30, 1988 



TITLE OF PROJECT (80 cnaracwrs or less. Title must til an one line oerween tne ooraers.i 

Clinical Slectrophysiology of the Visual System 



PRINCIPAL INVESTIGATOR (Usi otner orotassionai oersonnei oeiow tne Pnnaoai investigator. i (Name, title, /aooraro/v. ana institute artiliationi 



?I: Muriel I. Kaiser-Kupf er M.D. 



I 



Others: Rafael Caruso M.-D. 

Doris J. Collie A. A. 



Head, Section on Ophthalmic 
Genetics and Pediatric 
Ophthalmology 

Visiting Scientist 
Health Technician 



CB, NEI 



CB, NEI 
CB, NEI 



COOPERATING UNITS (it any/ 

Georgetown University Center for Sight, Washington, D.C. (Despina Koustsandr 
O.T., Robert Toma, C.O.T.) 



eas, 



; LAB/BRANCH 

t Clinical Branch 



SECTION 



I Section on Ophthalmic Genetics and Pediatric Ophthalmology 

INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN- YEARS; 
.60 



PROFESSIONAL; 

.30 



OTHER; 



.3 



CHECK APPROPRIATE BOX(ES) 

G (a) Human subjects 
n (a1) Minors 
G (a2) Interviews 



G (b) Human tissues 



(c) Neither 



SUMMARY OF WORK (Use stanaara unreaucea type. Do not exceaO tne space proviaea.) 

The visual function of patients with ocular diseases or lesions in the visual 
pathways and of normal subjects is measured objectively with electrophysio- 
! logical techniques. These data are correlated with those obtained with psycho- 
physical tests of visual function. The results obtained contribute to the 
diagnosis of ocular and neural disorders that affect vision, and are needed to 
characterize their nature and evolution. They are also valuable in the 
assessment of the effects of different forms of treatment on the outcome of 
these diseases. 



PHS 6040 (Rev 1/841 



SPO st«.«ia 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



Z01 EY 00211-03 CB 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (BO cnaraciers or less. Tim must til on one line Between me ooraers.) 

A Double-Masked Controlled Randomized Clinical Trial of Topical Cysteamine 



PRINCIPAL INVESTIGATOR (Ust otner proiessional personnel oeiow tne Pnnaoal Investigator.) (Name, title, laooraiory. ana institute atliliationi 



PI: Muriel I. Kaiser-Kupf er M.D. 



Others: Lessie McCain R,.N. 

Manuel Datiles M.D. 



Head, Section on Ophthalmic 
Genetics and Pediatric 
Ophthalmology- 
Clinical Technician 
Visiting Scientist 



CB, NEI 



CB, NEI 
CB, NEI 



COOPERATING UNITS (It any) 

Human Genetics Branch, NICHD, National Institutes of Health, Bethesda, Maryland 
(William Gahl, M.D., Ph.D.) 



LAB/BRANCH 

Clinical Branch 



SECTION 

Section on Ophthalmic Genetics and Pediatric Ophthalmology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS; 
.25 



PROFESSIONAL; 
.15 



OTHER; 



.1 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (al) Minors 
□ (a2) Interviews 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use stanOara unreaucetl type. Do not exceea me space proviaea.) 



Nephropathic cystinosis is an autosomal, recessively inherited storage disease 
in which nonprotein cystine accumulates within cellular lysosomes due to a 
defect in lysosomal cystine transport. Ocular manifestations include photo- 
phobia crystal deposits in cornea, conjunctiva, iris and depigmentation of the 
retina. Systemic complications include the Fanconi syndrome, and renal failure. 

'Eight years ago cysteamine, a free thiol which depletes cystine from cells, was 
introduced in the therapy of cystinotic patients. Although patients had 
improved growth and stabilized renal function, there was no noticeable effect 
on the accumulation of corneal crystals. Recent studies showed that corneal 
cells in tissue culture are readily depleted of cystine by the introduction of 
cysteamine, making feasible the use of topical ophthalmic cysteamine to circum- 
vent the humoral route. After appropriate animal studies to test for complica- 
tions which revealed none, we have begun a double-masked clinical trial to test 
the efficacy of topical cysteamine (0.1%) in humans. Sixteen patients have thus 
far been enrolled. Four patients have shown significant decrease in the 
cysteamine treated eyes and are now taking drops in both eyes. To permit 
increasing the concentration of cysteamine eye drops in humans, a study was 
performed in rabbits, permitting an increase in the concentration to 0.5$. 



PHS 6040 (Rev 1/84) 



SPO »^*•»\» 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



Z01 EY 00060-10 CB 



PERIOD COVERED 

October 1, 1987 to September 30. 1988 



TITLE OF PROJECT (80 cnancierz or less. Title must lit on one line oeiween tne oorxnrs.i 

Visual Function and Ocular Pigmentation in Albinism 



PRINCIPAL INVESTIGATOR (Ust otner oroiessionai oersonnei oeiow tne Pnnaoai investigator.) (Name, title, laooratory, ana institute atliliationi 

PI: Muriel I. Kaiser-Kupf er M.D. Head, Section on Ophthalmic 

Genetics and Pediatric 
Ophthalmology 



CB, NEI 



Others: Lessie McCain 
Rafael Caruso 



R.N. Clinical Technician 
M.D. Visiting Scientist 



CB, NEI 
CB, NEI 



COOPERATING UNITS (It any) 



LAB/BRANCH 

Clinical Branch 



SECTION 

Section on Ophthalmic Genetics and Pediatric Oohthalmnlngy 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
.65 



PROFESSIONAL 
.15 



OTHER. 



^3- 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects 
D (al) Minors 
n (a2) Interviews 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use stanaara unreauced rype. Oo not axcaati tne space proviaeO.) 



Patients with hypomelanotic disorders such as ocular albinism, oculocutaneous 
albinism, Chediak-Higashi disease, Hermansky-Pudlak syndrome, and iris 
transillumination defects are being recruited to determine visual function with 
these conditions and to evaluate its course over time. Family members are 
evaluated to attempt to determine factors which may identify the heterozygous 
state. 



PHS 6040 (Rev l/84> 



SPO ■I4.«1( 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



i PROJECT NUMBER 

! Z01 EY 00084-10 CB 



I PERIOD COVERED 

JDctober 1, 1987 to Septetnber 30 > 1988 



TITLE OF PROJECT (80 cnaraciars or mss. Tine must lit on one line oetween me ooraers.) 

interior Chamber Anomalies Associated with Glaucoma or Ocular Hypertension 



PRINCIPAL INVESTIGATOR (Ust omer oroiessionai oersonnei oeiow me Pnncioai invesugaior.) (Name, title. laDoratory. ana institute atliliationi 

PI: Carl Kupfer M.D. Director NEI 



Others: Muriel I. Kaiser-Kupf er M.D. 



Lessie McCain R.N. 

Manuel B. Datiles M.D. 
Paul Edwards M.D. 



Head, Section on 

Ophthalmic Genetics 

and Pediatric Ophthalmology 
Clinical Technician 
Visiting Scientist 
Visiting Fellow 



CB, NEI 



CB, NEI 
CB, NEI 
CB, NEI 



COOPERATING UNITS (if any) 



LA6/BHANCH 

Clinical Branch 



SECTION 

Section on Ophthalmic Genetics and Pediatric Ophthalmology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS; 
0.75 



PROFESSIONAL; 
0.55 



OTHER; 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects 
D (a1) Minors 
□ {a2) Interviews 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use sianaara unraaucaa type. Do not axceati tne space proviaea.) 



With recent erabryological research indicating the role of the neural crest in 
contributing to all connective tissues anterior to the lens epithelium, the 
group of developmental anomalies of the anterior chamber with glaucoma or 
ocular hypertension is being reviewed. 



PHS 6040 (Rev l/84> 



SPO «14-9I( 



ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October 1, 1987 - September 30, 1988 

REPORT OF THE CHIEF, LABORATORY OF IMMUNOLOGY 
Robert B. Nussenblatt, M.D. 



This was the third year for the Laboratory of Immunology, National Eye 
Institute. While the first year saw the establishment of four Sections within 
the Laboratory, each with its own Section Head, the end of the second year 
brought into the Laboratory a new Section, that of Molecular Biology. 
Therefore, at present the Laboratory's five Section Heads are: Section on 
Clinical Immunology, Alan G. Palestine, M.D., Section on Immunology and 
Virology, John J. Hooks, Ph.D., Section on Experimental Immunology, Igal Gery, 
Ph.D., Section of Molecular Biology, Toshimichi Shinohara, Ph.D., and Section 
on Immunoregulation, Robert B. Nussenblatt, M.D. 

Over the past year the Section on Clinical Immunology has been 
particularly interested in the question of possible autoimmunity to the 
anterior uvea in patients with uveitis. Though many forms of anterior uveitis 
are presumed to be due to autoimmunity, there has been no confirmation that an 
ocular specific antigen is indeed involved in this process. Patients with 
anterior uveitis have been screened for autoantibodies directly against bovine 
iris and antibodies have been detected in some patients to a protein with a 
molecular weight of approximately 22,000. This protein appears to be specific 
to the iris, and further studies are continuing in order to illustrate what 
might be the first identification of such an antigen in the anterior segment 
of the eye. Additionally, the section has been actively involved in the role 
of the neuro-endocrine axis on the immune response. While the section has 
previously shown that the use of bromocriptine, a prolactin inhibitor, can 
modulate S-antigen induced experimental autoimmune uveitis, this work has been 
carried into the human sphere. The results of a double-masked study using 
bromocriptine alone in an attempt to reduce the number of recurrent attacks of 
anterior uveitis demonstrated that there was no major difference between 
groups receiving this drug as opposed to placebo. Additionally, a second 
trial focuses on the additive effects of cyclosporine plus bromocriptine in an 
attempt to treat patients with posterior uveitis at lower dosages of 
cyclosporine in order to reduce its concurrent renal toxicity. These results 
continue to be collected with the very important evaluation of renal function 
to be done in the not too distant future. As well, the section has developed 
a variety of techniques to evaluate the role of the retinal vasculature in 
ocular inflammatory disease. This includes the growing of vascular 
endothelial cells as well as newer ways to evaluate the vasculature in vivo. 

The Section on Experimental Immunology has been actively involved in 
learning the pathogenesis of inflammatory eye diseases. They have 
concentrated particularly on the model for uveitis induced with ocular 
specific antigens. The focus of the past year has recently been on the 
interphotoreceptor retinoid-binding protein (IRBP) which is highly uveitogenic 
and produces experimental autoimmune uveitis (EAU) in various animals 
including primates. The focus has been on the identification of peptide 
determinants of IRBP that are responsible for inducing EAU and initiating 



immune responses. Some 14 peptides selected for synthesis from the IRBP 
sequence have been looked at and 4 peptides were found to be uveitogenic in 
Lewis rats. One of the peptides designated R14 was shown to be extremely 
potent in inducing disease at a dose as low as 0.0 6 micrograms per rat. 
Additionally, this protein as well as a second designated R4 were also found 
to produce EAU in primates. This work will be expanded over the ensuing year 
to evaluate the peptides from IRBP to which patients with severe inflammatory 
disease of the eye may respond. 

The Section of Molecular Biology over the past year has concentrated on 
the question of molecular mimicry as well determining the amino acid sequences 
of human, mouse and bovine S-antigen. Immunogenic sites, as well as 
uveitopathogenic sites, have been identified in this molecule. As with the 
IRBP molecule, EAU as well as pinealitis can be induced in Lewis rats with 
small peptides. Of interest, the disease can also be induced with a small 
peptide corresponding to the amino acid positions 10 6-117 in the yeast histone 
H3 which contains five consecutive amino acids identical to a uveitogenic 
pathogenic site in the human S-antigen. This potential cross-reactivity may 
provide us with future insight into basis mechanisms of cross-reactivity and 
molecular mimicry in the future. 

The Section on Immunology and Virology has continued to develop its 
interest into several areas including T-cell modulators . The development of 
virally induced diseases and also the study on the bioregulatory aspects of 
the retinal pigment epithelial (RPE) cell. The group has successfully 
modulated the expression of experimentally induced uveitis with the treatment 
of animals with anti-la therapy. Additionally, studies continue on the 
expression of class II antigens localized in autoimmune diseases both in human 
tissue as well as in the animal system. The presence of class II molecules on 
retinal blastema cells has also been demonstrated. The modulation of HLA-DR 
by gamma-interferon as well as the preferential expression of this determinant 
over HLA-DQ has also been shown. Of interest, double labeling studies have 
revealed that the HLA-DR antigen is shared concomitantly with cells of glial 
and neuronal character. The area of the retinal pigment epithelial cell has 
developed considerably over the past year. The group has identified two mouse 
IgG monoclonal antibodies which react with the human RPE cell. These 
monoclonal antibodies are both specific for the RPE cell within the eye and do 
not appear to react with any other ocular structures. Additionally, they do 
ijot react with human skin, kidney or peripheral mononuclear cells. These 
antibodies recognize cell surface molecules which are highly conserved since 
they can be found in not only man but also in monkey, rat, mouse, cow, chicken 
and frog. These are the first monoclonal antibodies which are directed solely 
at the human RPE cell. Additionally, the group has initiated studies to 
evaluate corona virus infections in the eye and optic nerve. These 
preliminary studies have begun with hope that these will come to fruition over 
the ensuing year. 



The Section on Immunoregulation has been evaluating the role of cytokines 
in human intraocular fluids . Intraocular fluids from patients who require 
surgery to repair a retinal detachment or surgery due to sequelae of uveitis 
have been evaluated. These patients' fluids were evaluated for the presence 
of interleukin-1 as well as interleukin-2 activity by bioassays . Ten p>ercent 



of uveitis patients, 20% of retinal detachment patients and 60% of patients 
with proliferative vitreal retinopathy (PVR) had detectable IL-2 activity. Of 
great interest was the fact that IL-1 activity was found in 90% of uveitic 
eyes, 35% of eyes with retinal detachment and 17% of eyes with proliferative 
vitreal retinopathy. IL-1 would seem to be a mediator in multiple organ 
specific pathways. Further, its presence in the eye suggests a role in 
intraocular inflammatory and immune processes and as well in ocular diseases 
that are not usually associated with the immune system. Of great interest was 
the relatively high percentage of PVR patients with IL-2 activity thus 
suggesting a role of the immune system in this proliferative vitreal 
retinopathy. The group has additionally begun the use of a new antibiotic, 
magainin. Preliminary studies in this area have demonstrated in vivo activity 
of magainin by showing a less severe corneal abscess in the treated animals 
with the delayed onset of the abscess as compared to the control animals. 
This important area will be continued over the ensuing year. The group has 
also used a molecular biologically prepared IL-1 linked to the exotoxin of 
pseudomonas . This novel approach has permitted the group to pinpoint cells 
that bear IL-2 receptors (such as activited clones that will be mediating 
uveitis) and destroy them. This experimental approach appears to be 
successful and will be looked at in greater detail over the ensuing year. 
Additionally, the mouse model of experimental autoimmune uveitis has been well 
developed and the development of long term cell lines as well as clones will 
be an important goal over the ensuing year. The therapeutic intervention in 
human intraocular inflammatory disease took an important step in the 
initiation of a phase 1/2 randomized trial using cyclosporine A and G. This 
study has great import in that cyclosporine G may be considerably less 
nephrotoxic and therefore may be a reasonable next generation 
immunosuppressive agent. 

The Laboratory of Immunology's Sections have produced significant 
observations over this past year, both clinically as well as from a basic 
research point of view. The goal for all is a better understanding of the 
basic mechanisms of ocular inflammatory diseases. This work will continue 
with this fabric of basic research combined with practical observations such 
as the treatment of patients with cyclosporine as well as other 
immunomodulating agents . 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



ZOl EY 00230-03 LI 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 
TITLE OF PROJECT (80 cfiaracmrs or lasa. Wa must tit on one una oatweon tna Oontars.f 

Modulation of Retinal Vascular Permeability by Inflammatory Mediators 



PRINCIPAL INVESTIGATOR (Ust otrrnr protaaamnai personnm Oalow trie Pnnaoal Invaaagator.) (Name, titta. laOorator/, ana institute allillation) 

PI: Alan G. Palestine M.D. Head, Section on Clinical 



Others: Rebecca Gurley 
David C. Herman 
Jeffrey N. Bloom 





Immunology 




M.S. 


Biologist 




M.D. 


Senior Staff 


Fellow 


M.D. 


Senior Staff 


Fellow 



LI, NEI 



LI, NEI 
LI, NEI 
LI, NEI 



COOPERATING UNITS (it any) 



LAS/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Clinical Immunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 
0.83 



PROFESSIONAL; 
0.43 



OTHER: 



0.4 



CHECK APPROPRIATE BOX<ES) 

D (a) Human subjects 
n (al) Minors 
n (a2) Interviews 



D (b) Human tissues El (c) Neither 



SUMMARY OF WORK (Un stanOara unraaucaa type. Oo not excaud tfia spaca pronOaa.) 

Retinal vascular leakage is an iinportant mechanism of visual loss in ocular 
inflammatory disease. The presumed site of retinal vascular leakage is the . 
retinal capillaries which are composed of pericytes and endothelial cells . 
Therefore, it is likely that immune mediated disease alters pericyte or 
endothelial function in a manner that produces vascular leakage. This project 
is concerned with quantifying the specific mediators that are involved in 
producing these changes so that more appropriate therapy can be targeted. 



PHS 6040 (Rav. 1/84) 



era tt44ti 



PROJECT NUMBER 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT i ^^^ ^^ 00217-03 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 cfiaractera or lass. Titla must tit an one line between ma ooraers.) 
Lymphocyte Migration in Experimental Autoimmune Uveitis 



PRINCIPAL INVESTIGATOR (Ust ottm protassmnal personnel Oelow me Rnnapal Investigator) (Name, title, labarator/ ansa inftituta attillation) 
PI: Alan G. Palestine M.D. Head, Section on Clinical li^ nEI 

Immunology 

Others: Robert B. Nussenblatt M.D. Clinical Director NEI 

Jeffrey N. Bloom M.D. Senior Staff Fellow LI, NEI 



COOPERATING UNITS (H any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Clinical Immunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.26 



PROFESSIONAL: 

0.26 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects D (b) Human tissues Q (c) Neither 

D (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Uau stanoanl unraOiKad typ». Oo not axcMd Ifta spaca prwiOKt.) 

Experimental autoiiranune uveitis (EAU) is induced by immunization of rats and 
other experimental animals with S-antigen (a soluble antigen from the retina) is 
being investigated in this laboratory as a model of human intra-ocular 
inflammation. This experimental inflammation can be transferred from donor rats 
to naive recipients using lymphocytes harvested from the spleen or lymph nodes. 
Following harvesting of the cells from the donors and three days in culture with 
stimulating antigen, the cells are injected into the intra-peritoneal cavity and 
five to seven days later the recipient rats develop EAU. The disease can also 
be transferred using a T-helper cell line by intra-peritoneal or intra-ocular 
injection. The mechanism of transfer of disease is unclear. This work has used 
radioactively labeled lymphocytes to determine the fate of these lymphocytes 
after injection into the peritoneal cavity or blood during the process of the 
development of uveitis. The goal of this project is to understand the 
initiating mechanisms of inflammation in the hope that these mechanisms can be 
extended and applied to human inflammations. Cells from an S-Ag specific T cell 
line migrate into the retina and cause EAU. The kinetics of this migration are 
being studied. S-antigen specific cells reach the eye in greater numbers if the 
inflammation in the eye is induced by S-antigen than if it is induced by another 
mechanism. 



PHS 6040 (Rav. 1/84) awot\*-*ta 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBES 

ZOl EY 00218-03 LI 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (BO efmrmctmfs or mst. TiO» must tit on on* WW Miw**n tnt oomtn.) 

Acquired Immune Deficiency Syndrome 



PRINCIPAL INVESTIGATOR (Lat ottmr prolmtnonm oersonnn OHow tnt PrmcttU imtttagamr.) (Ntnrn. tnm. luoormtory, ana msmun attumoon) 

PI: --^lan G. Palestine M.D. Head, Section on Clinical LI, NEI 

Immunology 



Others: 



Robert B. Nussenblatt 



M.D. 



Clinical Director 



NEI 



COOPERATING UNrrs f/r an)-; Laboratory of Tumor Cell Biology, National Cancer Institute 
(S. Zaki Salahuddin, Ph.D.); Laboratory of Cellular & Ilolecular Biology, National 
Cancer Institute (Dharam Ablashi, D.V.M.); Department of Critical Care Medicine, 
Clinical Center (Henry Masur. M.D.): Laboratory of Tumor Cell Rinlntfv. Nafinnal 

LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Clinical Immunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.09 



PROFESSIONAL: 



0.09 



OTHER; 



CHECK APPROPRIATE BOX(ES) 

[1 (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



D (b) Human tissues D (c) Neither 



SUMMARY Of WORK (Ut» sonoafS ummaucue typt. Do not momA vm spie* pmnaaO.} 

Cytomegalovirus retinitis is the major cause of blindness in AIDS patients. 
Although we have previously shown that ganciclovir is effective in treating this 
infection, the disease relapses without continued maintenance. Maintenance 
therapy requires intravenous infusion and is associated with marrow toxicity. A 
multi-center randomized trial is currently being planned to evaluate the use of 
this drug. 



PHS 6040 (Rev 1/84) 



S^O •!<••<• 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 00219-03 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (30 ctianctars or msi. 7708 must tit on one line oeiwaen ma oaraen.l 

The Effect of Bromocriptine on Human Uveitis 



PRINCIPAL INVESTIGATOR (List offw protaasionw oersonnm Oelow ma Pnncipal Investigator.) (Name, atte. /aoorafcwy. ana instnuta attiliaoan) 

PI: Alan G. Palestine M.D. Head, Section on Clinical LI, NEI 



Others: Robert B. Nussenblatt M.D. 

Janet L. Davis M.D. 

David C. Herman M.D. 

Jeffrey N. Bloom M.D. 



Immunology 

Clinical Director 
Senior Staff Fellow 
Senior Staff Fellow 
Senior Staff Fellow 



NEI 
LI, NEI 
LI, NEI 
LI, NEI 



COOPERATING UNITS (it any) 

Metabolism Branch, National Cancer Institute (Marie C. 



Gelato, M.D.) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Clinical Immunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 

0.91 



PROFESSIONAL 

0.91 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

El (a) Human subjects 
n (a1) Minors 
n (a2) Interviews 



D (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Uam stanoant unnauead typa. Oo not axcmad tfia apaoa promOaa) 



In recent years there has been increasing evidence in the literature that 
pituitary hormones are capable of regulating the immune system. There is 
evidence to suggest that prolactin is an immunostimulatory hormone and that 
reduction of serum prolactin levels in experimental animals by hypothesectomy or 
treatment with bromocriptine will result in a degree of immunosuppression. 

This information has been applied to hvunans and two clinical studies have begun. 
Both of these are in early phase of patient recruitment. One study is a 
randomized trial between placebo and bromocriptine in recurrent anterior uveitis 
using the end point of the number of recurrences per year to determine whether 
bromocriptine is capable of regulating the immune system in these patients . The 
second trial focuses on the additive effects of cyclosporine plus bromocriptine 
in attempts to treat patients with posterior uveitis at lower doses of 
cyclosporine to reduce its concurrent renal toxicity while at the same time 
achieve an immunosuppressive effect . Cyclosporine and prolactin compete for 
binding sites on the lymphocyte. 

Further studies in human disease will hopefully elucidate other aspects of the 
neuroendocrine axis which can be utilized to regulate the immune system to treat 
autoimmune diseases . 



PHS 6040 (Rov. 1/84) 



sro Il4**<t 



PROJECT .MUMSER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT i ^^^ ^"^ 00220-03 LI 



I PERIOD COVERED 

I October 1, 1987 to September 30, 1988 

i TITLE OF PROJECT (80 cnaracmrs or lass. Tlda must tit on one line Oetwean me txiraers.) 

I Endocrine Modulation of Immune-Mediated Eye Disease in Rats 



PRINCIPAL INVESTIGATOR (Ust other professional personnel oeiow me Pnnapal Investigator.) (Name. ava. laOoratory, ana institute atfillaHon) 
PI: Alan G. Palestine M.D, Head, Section on Clinical LI, NEI 

Immunology 

Others: Robert B. Nussenblatt M.D. Clinical Director NEI 

David C. Herman M.D. Senior Staff Fellow LI, NEI 



COOPERATING UNITS (U any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Clinical Iiranunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEAHS: 
0.31 



PROFESSIONAL: 
0.31 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects D (b) Human tissues Q (c) Neither 

D (al) Minors 
D (a2) Interviews 



SUMMARY OF WORK CUM stanaairi unretiucaa type. Do not excaml (/)• space prwided.) 

In recent years there has been increasing evidence in the literature that 
hormones are capable of regulating the immune system. There is evidence to 
suggest that prolactin is an immunostimulatory hormone and that reduction of 
serum prolactin levels in experimental animals by hypotheaectomy or treatment 
with bromocriptine will result in a degree of immunosuppression. 

An animal model of experimental autoimmune uveitis (EAU) induced by immunization 
of rats with S-antigen (a soluble antigen from the retina) is used to study 
intraocular inflammatory disease. We have demonstrated a decrease in antibody 
production in both male and female rats and a decreased incidence of uveitis in 
female animals but no significant effect on the immune responses measured by 
lymphocyte proliferation. As reported before, high doses of cyclosporine (10 
mg/kg) results in only partial reduction of intraocular inflammation. We have 
demonstrated that the suppression of prolactin by concurrent use of bromo- 
criptine in combination with low dose cyclosporine is more effective than either 
drug separately in suppressing both the incidence of disease as well as cellular 
and humoral immune responses . Evidence in the literature suggests that 
cyclosporine competes with prolactin for binding sites on lynphocytea therefore 
reductions in prolactin level may reduce competition for those sites and make 
cyclosporine treatment more effective. Further studies with this animal model 
will elucidate other aspects of the neuroendocrine axis that may be utilized to 
I regulate the immune system to treat autoimmune diseases . 



PHS 6040 (R«v. 1/84) awo ti4-»i* 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 00221-03 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 charucmrs or tesa. Title must M on one line Oerween ttie ooraers.) 
Intraocular Class II Antigen Expression in Endotoxin-Induced Uveitis 



PRINCIPAL INVESTIGATOR (List otfmr protaaamnal personnel Oelow the Pnncipal Investigator.) (Name, title, laooratoty, ana institute alfiliaoon) 
PI: Alan G. Palestine M.D. Head, Section on Clinical 

Immunology 



LI, NEI 



Others: Robert B. Nussenblatt M.D. 
Jeffrey N, Bloom M.D. 
David C. Herman M.D. 



Clinical Director 
Senior Staff Fellow 
Senior Staff Fellow 



NEI 
LI, NEI 
LI, NEI 



COOPERATING UNITS (it any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Clinical Immunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.51 



PROFESSIONAL; 
0.51 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



D (b) Human tissues El (c) Neither 



SUMMARY OF WORK (Use atanOarti unnoucad type. Do not axcaad the space provtOea.) 

Endotoxin is a polysaccharide derived from the cell wall of gram negative 
bacteria. When injected, into the footpad or the eye of a rat it will induce an 
inflammatory reaction within the eye. The mechanism of this inflammation is 
still unclear. However, since several types of anterior uveitis in humans 
appear to be linked to gram negative bacteria exposure, this is considered a 
relative model for anterior uveitis in humans such as Reiter's syndrome. In 
this study the expression of class II antigens was studied within the eyes of 
rats receiving E. coli endotoxin by immunohistochemical techniques. We observed 
that the expression of class II antigens on the ciliary body and iris proceeded 
the influx of inflammatory cells into the eye and that the inflammatory cells 
that entered the eye- were primarily neutrophils with some monocytes . No T-cells 
were present in the inflammatory infiltrate. The inflammatory cellular 
infiltrate could be inhibited by indomethacin or colchicine, however this did 
not alter the expression of class II antigens by the iris or ciliary body 
indicating that this expression is not singly a consequence of the inflammatory 
infiltrate but may be intimately involved with the mechanism of the expression 
of endotoxin induced uveitis. Corticosteroids were capable of suppressing both 
the cellular inflammatory infiltrate and the expression of class II antigens. 
The expression of class II antigens on nonlymphoid cells within the eye may be 
important in antigen presentation or may simply signal a phenotypic change on 
the cells due to the interaction of endotoxin with the cell membranes. The 
findings were compared with the expression of class II antigen in passive and 
active intraocular Arthus. The effect of endotoxin on ocular inflammation was 
studied using fluorophotometry to validate the use of animal studies as a useful 



PHS 6040 (Rev. 1/84) 



SPO (I***!* 



DEPARTMENT OF HEALTH AND HUMAN SERVICES ■ PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT MUVBER 



ZOl EY 00247-01 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or *m. TWe must M on one line between the boraers.) 
Autoimmunity to the Anterior Uvea in Patients with Uveitis 



PRINCIPAL INVESTIGATOR (List other protesaionU personnel below tfie Pnnapal Investigator.) (Name, title, laboratory, ami mstituta affiliation) 
PI: Alan G. Palestine M.D. Head, Section on Clinical LI, NEI 

Immunology 



Others; 



Rebecca Gurley 



M.S. 



Biologist 



LI, NEI 



COOPERATING UNITS (If any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Clinical Immunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 
0.77 



PROFESSIONAL: 

0.17 



OTHER: 



0.6 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



[^ (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use atandara urueducud lypa. Do not exceed Ute space provided.) 

Many forms of anterior uveitis are presumed to be due to autoimmunity directed 
towards ocular antigens. However, there has been no confirmation that an 
ocular specific antigen is involved in this process. It is important to 
develop an understanding of the mechanisms of inflammation in patients that 
have anterior uveitis. The presumed site of inflammation in these patients is 
the iris and ciliary body. We, therefore, began to look for iris specific 
proteins to which patients might have an autoimmune response. Patients with 
.anterior uveitis were screened for auto-antibodies directed against bovine 
iris. Antibodies were detected to a protein with a molecular weight of 
approximately 22,000 in some patients. When compared to a control group, 
patients, in general, have higher levels than control individuals of this 
antibody. Until the protein is isolated and T-cell responses can be measured, 
the true significance of these antibodies will be unclear. Antibodies to 
retinal antigens are much less revealing than the corresponding T-cell 
responses in distinguishing patients from controls. The protein that has been 
identified appears to be specific to the iris and is not found in other 
tissues of the body. Purification of this protein for other immunologic 
studies is in progress . 



PHS 6040 (Rev. 1/84) 



CPO t14.«ll 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 00069-11 LI 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PHOJECT (30 cfiaracnrs or less. Title must tit on one line tiecween tne Ooraers.) 
Immune Responses to Ocular Antigens 



PRINCIPAL INVESTIGATOR (List otfw prolesaianal personnel Oelow me Principal Investigator.) (Nairn, title, laboratory, ana insottita atfillaoon) 
PI: Igal Gery Ph.D. Head, Section on Experimental LI, NEI 

Immunology 



Others: Shigeto Hirose M.D. 

Hiroki Sanui M.D. 

Takao Tanaka M.D. 

LiHong Hu M.D. 

Satoshi Kotake M.D. 



Visiting Fellow 
Visiting Associate 
Visiting Fellow 
Visiting Fellow 
Visiting Fellow 



LI, NEI 

LI, NEI 

LI, NEI 

LI, NEI 

LI, NEI 



COOPERATING UNITS (if arty) 

Stephen I. Katz 
Hanah Margalit 
Horst W. Korf 



M.D. 
Ph.D. 



DB, NCI, NIH 
LMB, NCI, NIH 



University of Geissen, FRG 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Experimental Immunology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 
7.83 



PROFESSIONAL: 

7.43 



OTHER: 



0.4 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
n (a2) Interviews 



D (b) Human tissues E (c) Neither 



SUMMARY OF WORK (Usm standard unreduced type. Do not exeaed ttm spact prmtdad.) 

This project is aimed at learning about the pathogenesis of inflammatory eye 
diseases which are grouped under the term "uveitis". As a model for uveitis 
in man we have induced "experimental autoimmune uveoretinitis" (EAU) in 
experimental animals by immunization with ocular-specific antigens. We have 
recently shown that a retinal component, the interphotoreceptor 
retinoid-binding protein (IRBP) is highly uveitogenic and produces EAU in 
various animals, including primates. Our main effort in FY-1988 has focused 
on the identification of peptide determinants of IRBP that are responsible for 
inducing EAU and initiating immune responses. Of the fourteen peptides 
selected for synthesis from the IRBP secpaence, four peptides were found to 
induce EAU in Lewis rats. One of the peptides, designated R14, was extremely 
potent, inducing disease at a dose as low as 0.06 |J,g/rat; the other three 
peptides were approximately 1000 fold less active. A correlation was found 
between the capacity of the peptides to induce EAU and to initiate cellular 
immunity which cross reacts with the native IRBP molecule. Two peptides, R4 
and R14, were also found to produce EAU in primates, thus suggesting that 
these peptides could be involved in htiman uveitio conditions as well. 

In other studies we have collected data to suggest the possible involvement of 
non-MHC- restricted killer lymphocytes ("NK" and "LAK") and of interferon-Y 
(IFN-y) in the pathogenesis of EAU. A marked increase in the 
non-MHC-restricted cytotoxic activity was observed in monkeys immunized for 
induction of EAU. Treatment of mice with IFN-y significantly elevated the 
expression of la (class II) antigens on various ocular cells, with a pattern 
resembling that seen in animals which develop EAU. 



PHS 6040 (Rav. 1/84) 



a'o ti4-»i* 



^ 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

i ZOl EY 00232-03 LI 
NOTICE OF INTRAMURAL RESEARCH PROJECT I 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must tit on one line between the txjrders.) 
Interferon System in Cellular Function and Disease 



PRINCIPAL INVESTIGATOR (Ust other professions personnel below the Principal Investigator) (Name, title, laboratory, ana institute affiliation) 

PI: John J. Hooks Ph.D. Head, Section on Immunology LI, NEI 

and Virology 

Others: Barbara De trick Ph.D. Expert LI, NEI 

Caroline Percopo B.S. Biologist LI, NEI 

Christian Hamel M.D. Visiting Fellow LI, NEI 

Muriel Kaiser-Kupfer M.D. Head, Section on Ophthalmic CB, NEI 

Genetics 



COOPERATING UNITS (it any) 

Jan Vilcek M.D. New York University, School 

of Medicine 
Charles Evans M.D. Head, Tumor Biology Section LB, NCI 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunology and Virology 



INSTmjTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
1.1 



PROFESSIONAL: 

0.7 



OTHER: 

0.4 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects D (b) Human tissues ^ (c) Neither 

n (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Ust stanOart) unreOuced type. Do not sxcMd the apaca provided.) 

The interferon (IFN) proteins can modify a variety of biological activities 
and are considered one of the body's regulatory proteins. Numerous studies 
now indicate that the IFN's are potent immunoregulators . During the past year 
we have been studying the ways in which IFN proteins interact with cells of 
the immune system and how this interaction may modify immune responses and 
immunologically related disorders . 

Using immunocytochemical analysis we have developed a sensitive method of 
'identifying lymphokines, IFN-gamma and IL2, at the site of tissue damage. We 
have identified these lymphokines in inflammatory eye diseases . The presence 
of these lymphokines is associated with a lymphocyte infiltrate predominantly 
of a T-cell origin and with the expression of MHC class II antigens on both 
the infiltrating cells and in the retinal pigment epithelial (rpe) cells. 

Experimentally we have shown that this direct intravitreal inoculation of 
recombinant rat IFN-y results in the expression of MHC Class II antigen (la) 
in a variety of ocular cells . 

This is the first demonstration of lynphokines , IFN-gamma and IL2 at the site 
of a localized autoimmune disease. These observations may indicate that 
IFN-gamma induced MHC class II antigen expression may serve as a local 
an?Jlification system in autoimmune and inflammatory eye disease. A better 
understanding of the role of lymphokines in the mechanisms involved in the 
development of autoimmunity and inflammation may be beneficial in the 
treatment of these diseases. 



PHS 6040 (Rev. 1/84) SPO •t4-«ia 



DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



PROJECT NL/MBER 

ZOl EY 00233-03 LI 



TITLE OF PROJECT (BO characters or less. Title must lit on one line Ixtween ttie txiroers.) 

Studies on the Bioregulatory Aspects of the Retinal Pigment Epithelial Cell 



PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute atlillatlon) 

PI: John J. Hooks Ph.D. Head, Section on Immunology LI, NEI 

and Virology 



Others: Barbara Detrick Ph.D. 
Caroline Percopo B.S. 
Susan Robbins Ph.D. 
Laura Caspers-Velu M.D. 
Shuji Suzuki M.D. 



Expert 
Biologist 

Postdoctoral Fellow 
Visiting Associate 
Visiting Associate 



LI, NEI 

LI, NEI 

LI, NEI 

LMOD, NEI 

LI, NEI 



COOPERATING UNITS (If any) 

Lawrence Bowsell M.D. 

Alain Bernard M.D. 

Reuben Siraganian M.D. 



Hopital St. Louis, France 
Institute Gustave Rowsse, France 

NIDR, NIH 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunology and Virology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
1.96 



PROFESSIONAL: 

1.76 



OTHER: 



0.2 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
n (a2) Interviews 



El (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standard unraducad type. Do not axeaad the apaoa prwidad.) 

The retinal pigment epithelial (rpe) cell is a major regulatory cell in the eye. 
That is, the rpe cell exerts a variety of actions in maintaining retinal 
integrity and function. In order to more effectively study this cell in vivo 
and in vitro, we have produced monoclonal antibodies directed against human rpe 
cells. 

Using immunoperoxidase assays (ABC) , we have identified two mouse IgG monoclonal 
antibodies which react with the human rpe cell. The monoclonal antibodies are 
both specific for the rpe cell within the eye, since they do not react with any 
other ocular structures. Moreover, these antibodies do not cross react with 
human skin, kidney or peripheral mononuclear cells. These antibodies recognize 
cell surface molecules which are highly conserved since they can be found in 
man, monkey, rat, mouse, cow, chicken and frog. 

These are the first monoclonal antibodies which are directed solely at the human 
rpe cell. Further characterization and studies with this antibody should prove 
useful in the identification of rpe cells in situ and in vitro. Moreover, this 
immunoglobulin will allow us to probe the bioregulatory functions of the cell. 



PHS 'r'Hn (Rp>' I""' 



GPO 01 rf-P! 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



=boject number 



ZOl EY 00234-03 LI 



PEFIIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 cttaracMrs or lass, fitle must tit on one line Oorwean tne boraers.) 
MHC Class II Antigens in the Pathogenesis of Inflanmatory Diseases 



PRINCIPAL INVESTIGATOR (Ust OBW protassionat personnel Delow trie Pnnapal Investigator) (Name, title. laOoratory, ana insotuta amiiatmn) 
PI= -John J. Hooks Ph.D. Head, Section on Immunology LI, NEI 

and Virology 



Others: Barbara Detrick Ph.D. 

Caroline Percopo B.S. 

Chi-Chao Chan M.D. 

Robert B, Nussenblatt M.D. 



Expert 
Biologist 
Medical Officer 
Clinical Director 



LI, NEI 
LI, NEI 

LI, NEI 
NEI 



COOPERATING UNITS <H any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Immunology and Virology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.40 



PROFESSIONAL- 

0.30 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
n (a2) Interviews 



OTHER: 



0.1 



D (b) Human tissues EI (c) Neither 



SUMMARY OF WORK (Use staiKtarO unraOucaa typm. Oo not ex ca eH the apaem proindatl.) 



MHC class II antigens, HLA-DR in the human and la in the mouse, are membrane 
bound glycoproteins that are encoded by genes of the major histocompatibility 
complex. Expression of these antigens is of great functional importance for 
the initiation and perpetuation of immune responses. In a number of immuno- 
pathologic conditions HLA-DR antigen negative cells are stimulated to express 
class II antigens. In these cases an immunologic role has been postulated for 
the class II antigen expression. 

During the past year, we have determined if class II antigens are expressed in 
certain diseases as well as evaluated their possible role in autoimmune and 
inflammatory diseases. Initial studies identified cells in the anterior 
segment and cells in the retina (rpe cell) which express class II antigens 
during inflammatory eye diseases. Treatment with monoclonal anti-la 
antibodies diminished the clinical disease and the expression of MHC class II 
antigens . 

These studies on MHC class II antigen expression in localized autoimmune 
diseases provide evidence that the activation of these antigens may contribute 
to the immunopathogenesis of these diseases. 



PHS 6040 (Rav. 1/84) 



aro it4.«i* 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 00240-02 LI 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. We must tit on one line between the txjrders.) 
Virus Infections in the Eye 



PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name. tnle. laboratory, and institute attillation) 

PI: John J. Hooks Ph.D. Head, Section on Inmunology LI, NEI 



Others: 







and Virology 


Susan Robbins 


Ph.D. 


Postdoctoral Fellow 


Christian Hamel 


M.D. 


Visiting Fellow 


Barbara Detrick 


Ph.D. 


Expert 


Caroline Percopo 


B.S. 


Biologist 


Robert B. Nussenblatt 


M.D. 


Clinical Director 



LI, NEI 
LI, NEI 
LI, NEI 
LI, NEI 
NEI 



COOPERATING UNITS (H any} 

See attached 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunology and Virology 



INSTITUTE AND LOCATION 

NEI. NIH. Bethesda. Maryland 20892 



TOTAL MAN-YEARS: 
1.00 



PROFESSIONAL; 



0.90 



OTHER: 



0.1 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



El (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standmni uni9duc0d type. Do not cxcMd the space provtdad.) 

During the past year we have initiated studies to evaluate the various 
virologic and immunopathologic processes which occur when viruses replicate in 
the ocular microenvironinent . This is a new project which is composed of three 
areas: (1) Evaluation of virus spread in HSV-1 induced retinitis. (2) 
Studies on coronavirus infection in ocular and optic nerve cells. (3) 
Determination of possible role of other viruses in hvunan eye diseases . 

Retinitis following anterior chamber inoculation of herpes siii^jlex virus 
(HSV-1) is an interesting model of viral spread and virus induced disease. 
During the past year we have elucidated some of the pathologic mechanisms 
involved in this disease. We found that footprints of the immune system 
(IFN-gamma and MHC class II antigen expression) can be identified in the 
protected retina strongly indicating that it is the immune system which 
protects the retina from virus destruction. Moreover, we identified the virus 
in the capillary body and ciliary nerves suggesting that this may be the mode 
of spread of the virus to the uninjected eye. Elucidation of virus spread and 
activation in the retina may provide insight into these same mechanisms in 
human disease, such as acute retinal necrosis. 

We have initiated studies to evaluate coronavirus infections in the eye and 
optic nerve. Monoclonal anti-virus receptor antibody has identified selected 
cells within the eye which express virus receptors . Preliminary studies 
indicate that the virus is capable of inducing ocular damage in the posterior 
pole. 



PHS wyn io_. 



r. I'c o 1 4-i> t » 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBES 

ZOl EY 00184-06 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 cnaractars or less. We must M on on« urn Oerwean me Ooraers.) 
Cellular Mechanisms in Uveitis 



PRINCIPAL INVESTIGATOR (Ust oV»r profsannai parsonnm Oeiow me Phnapal Investigator.) (Name, ooe, laDoratorY. anO institute atfUlation) 
PI: Rachel R. Caspi Ph.D. Visiting Associate LI, NET 



Others: Robert B. Nussenblatt M.D. 

Francois Roberge M.D. 

Chi-Chao Chan M.D. 

William Leake M.S. 

Makoto Higuchi M.D. 



Clinical Director 
Visiting Associate 
Medical Officer 
Biologist 
Visiting Fellow 



NEI 
LI, NEI 
LI, NEI 
LI, NEI 
LI, NEI 



COOPERATING UNITS (It any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTTTUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
2.06 



PROFESSIONAL: 

2.02 



OTHER: 



0.04 



CHECK APPROPRIATE BOX(ES) 

G (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



G (b) Human tissues [^ (c) Neither 



SUMMARY OF WORK (Uaa ttandartl unraOucad (ypt. Do not excauO the spact provided.) 

Cellular mechanisms of ocular iinmunologically-mediated disease are being studied 
in animal models of experimental autoimmune uveoretinitis . For this purpose, 
previously established models are used (eg, S-Ag uveitis in the Lewis rat) and 
new models are being developed (IRBP and S-Ag uveitis in different strains of 
mice) . In vivo-functional long-term T-cell lines and T-cell clones are 
developed and maintained in vitro from lymphoid organs of experimental animals 
immunized with uveitogenic ocular proteins. The phenotype and functional 
properties of these cells, as well as their interaction with ocular resident 
cells are being studied. The goal of these studies will be to identify the 
immunoreactive cells and mediators involved in the intraocular inflammatory 
process. 



PHS 6040 (Rev. 1/84) 



6Pe etfta 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT .MUMSER 



I 



ZOl EY 00222-03 LI 



PEHIOD COVEHED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (30 c/ianaars or lesa. Wa must ht on ona una OeMean me ooraors.; 

Immunopathology in the Eyes with Experimental Autoimmune Uveitis (EAU) 



PRINCIPAL INVESTIGATOR (Ust oWar pramssionai personnel Oelow me Pnnapal Investigator.) (Name. twa. laOoratoiy, ana institute affiliation) 
PI: Chi-Chao Chan M.D. Medical Officer LI, NEI 



Others: Robert B. Nussenblatt M.D. 

Igal Gery Ph.D. 

Rachel R. Caspi Ph.D. 

Francois Roberge ' M.D. 



Clinical Director NEI 

Head, Section on LI, NEI 

Experimental Immunology 

Visiting Associate LI, NEI 

Visiting Associate LI, NEI 



COOPERATING UNITS (It any) 

University of Tokyo, School of Medicine (Manabu Mochizuki, M.D.) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 
0.31 



PROFESSIONAL: 

0.31 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



D (b) Human tissues \B (c) Neither 



SUMMARY OF WORK (Uam stanoant unreOueaa type. Oo not axcmml the apaeu promOaAf 

Identity and topographic localization of immunocompetent cells and alteration of 
surface markers on ocular resident cells in rodents with experimental autoimmune 
uveoretinitis by active immunization or adoptive transfer were analyzed by 
immunohistochemical studies. The lymphocyte population at the inflammatory 
sites was found to change markedly during the course of disease. In the early 
stage, T-helper/ inducers are the predominant cells in the eye. A relative 
increase of T-suppressor/cytotoxic cells in the late stage were observed. 
Expression of major histocompatibility complex class II antigens on ocular 
resident cells such as RPE, retinal endothelium, keratocytes, fibroblast and 
ciliary epithelium was observed in different models of EAU in rats. This 
antigen expression may play a certain role in the pathogenesis of EAU. Both 
infiltrating (cell subpopulation) and expression of class II antigens on ocular 
resident cells, can be modulated by different immunosuppressive agents. 

Dynamics of EAU induced by adoptive transfer of S-antigen-specific T-lynphocyte 
cell line has showed that this cell line can recognize the photoreceptor S-Ag in 
vivo. Immunopathology in the eyes with EAU in mice can be presented as a 
chronic granulomatous inflammation. Development of sutoretinal neovasculariza- 
tion may occur. Expression of major histocompatibility complex class II 
antigens is only located on ocular resident cells with the presence of 
inflammatory cells. 



PHS 6040 (Rav. 1/84) 



OPO tt4-*ta 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



i=ROjECT .'JUMBER 

ZOl EY 00224-03 



LI 



PERIOD COVERED 

October 1, 1987 to September 30, 



1988 



TITLE OF PROJECT (30 ctiaracmrs or lass. Title must tit on one line tjetween mo Ooraars.) 
Sympathetic Ophthalmia: Immunopathclogical Findings 



PRINCIPAL INVESTIGATOR (Ust ottitr protaasKmai personnel OoWw me Pnnapal Investigator./ (Name, title, laooratory, and institute altiliaoan) 



PI: Chi-Chao Chan 

Others: Robert B. Nussenblatt 
Alan G. Palestine 

Toichiro Kuwabara 



M.D. 

M.D. 
M.D. 

M.D. 



Medical Officer 

Clinical Director 

Head, Section on Clinical 

Immunology 
Head, Laboratory of 

Ophthalmic Pathology 



LI, NEI 



LI, 
LOP, 



NEI 

NEI 

NEI 



COOPERATING UNITS (H any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTTTUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.12 



PROFESSIONAL: 



0.12 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (al) Minors 
D (a2) Interviews 



\2 (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use stanOai^ unreOucad type. Oo not excaea tfie space provnjaa.) 



Immunocompetent cells and ocular resident cells in the ocular tissues from 
patients with a clinical diagnosis of sympathetic ophthalmia were examined using 
the iinmunohistochemical technique. The choroidal infiltrates were composed 
primarily of T-lymphocytes . Different amounts of macrophages and B lymphocytes 
were present in each case. A varied spectrum of iitimunopatho logical and 
histopathological findings may occur in clinically diagnosed sympathetic 
ophthalmia. The immunopathology resembles EAU induced by retinal soluble model. 
Exposure of uveal tissue outside the eye and adjuvant effect may be important in 
the pathogenesis of this disease in humans . 



PHS 6040 (Rav. U84) 



SPO •I4-*I« 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 00225-03 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 ctiancters or lass. TitlB must lit on one Una Oetween tna ooraars.) 

Post- Inflammatory Complications in Uveitis 



PRINCIPAL INVESTIGATOR (List oOTw prorassional personnai OeH3w ma Principal Investigator.) (Nama, trtta. lat>orator^. ana institutB attillation) 
PI: Chi-Chao Chan M.D. Medical Officer 



Others: Robert B. Nussenblatt M.D, 
Francois Roberge M.D. 



Clinical Director 
Visiting Associate 



LI, NEI 

NEI 
LI, NEI 



COOPERATING UNITS (if any} 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 
0.12 



PROFESSIONAU 

0.12 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



Q (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Us» staMara unraOucad typa. Oa not axcamt Vm space prmiaa.) 



Complications of post-inflammation in uveitis patients includes destruction of 
photoreceptors, gliosis, choroidal scar, and formations of cyclitic membrane, 
snowbanking and preretinal membrane. Pos-inflammatory membrane composition may 
play an important role in the cause of complications associated with uveitis. 
In this study, eyes enucleated from patients with end stages of chronic anterior 
uveitis (formation of cyclitic membrane) , pars planitis (formation of preretinal 
membrane) were evaluated imraunohistochemically. Glial cells and proliferating 
Mialler cells were the major cellular components in these membranes. Basement 
membrane-like components and new collagens were the major extracellular membrane 
components . 



PHS 6040 (Rav. 1/84> 



ere ti4**ii 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMsea 
ZOl EY 00226-03 LI 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (SO cnaracmrs or lasa. fif/o must tit on ona lina oatween ine Oomers.) 

Immunopathology of Ocular Onchocerciasis and Other Parasitic Diseases 



PRINCIPAL INVESTIGATOR (List ot/Jar prorassional pefsonna/ Oo/ow tha Pnncipal Invaaagatar.) (Noma. Ma. laOoratory, ana insatuta attillation) 

PI: Chi-Chao Chan M.D. Medical Officer LI, NEI 



Others: 



Robert B. Nussenblatt M.D. 



Clinical Director 



NEI 



COOPERATING UNITS (if any) 

National Institute of Allergy and Infectious Diseases, Clinical Parasitic 

Diseases Section (Eric A. Ottesen, M.D.); World Health Organization (K. Awadzi, 

M.D.) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 

0.33 



PROFESSIONAL: 

0.33 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (al) Minors 
n (a2) Interviews 



B (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Usa aundai^ unnOucad typm. Oo not •xcaad ttm apacm prxivtaml.) 

Ocular specimens and sera from 12 patients with onchocerciasis and 10 controls 
were studied. A mild to moderate chronic inflammatory cellular infiltration was 
present in the conjunctiva of the onchocerciasis patients . T-lymphocytes were 
the predominant inflammatory cells with the T-suppressor subset being 
significantly increased in the onchocerciasis patients when compared to 
controls. In the onchocerciasis patients, the nonlymphoid cells in the 
conjunctiva and iris, such as vascular endothelia, pericytes and fibroblasts, 
showed an increase in expression of class II antigens. The anti-Onchocerca 
volvulus antibodies in the sera and aqueous humor were significantly higher in 
the patients compared to the controls . These findings suggest that T-cells are 
important in the ocular immune response to Onchocerca and that expression of 
class II antigens on nonlymphoid cells and the humoral factors may all play a 
critical role in ocular onchocerciasis . 

Retinal auto-antibodies in sera of these 12 patients were found. They were 
bound to the inner retinal layer and photoreceptors . Such autoimmune antibodies 
may play a role in the pathogenesis of the retinal degeneration and optic 
atrophy that occurs as a consequence of onchocerciasis . 



PHS 6040 (Rav. 1/84) 



e^o •i44tt 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT MUMsea 
ZOl EY 00241-02 LI 



PERIOD COVERED 
October 1, 



1987 to September 30, 1988 



TITLE OF PROJECT (80 cnancters or lasa. Title must tit on one line Oetween me ooraen.) 
Inununopathology of Ocular Diseases 



PRINCIPAL INVESTIGATOR (List ottier protassional personnel Oetow the Principal Investigator.) (Name, title. laOoratory. ana instittM attillaoon) 
PI: Chi-Chao Chan M.D. Medical Officer LI, NEI 



Others: Robert B. Nussenblatt M.D. 

Alan G. Palestine M.D. 

Ming Ni M.D. 

Toichiro Kuwabara M.D. 



Clinical Director 

Head, Section on Clinical 

Immunology 
Visiting Fellow 
Head, Laboratory of 

Ophthalmic Pathology 



NEI 

LI, NEI 

LI, NEI 
CB, NEi 



COOPERATING UNITS ^*any> zhongshan Ophthalmic Center, Guangzhon, Chine (Winifred Mao, 
M.D.); University of Iowa (Jay H. Frachmer, M.D.); Georgetown University Center 
for Sight (Michael Lemp, M.D.) 



UVa/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Immunoregulation 



INSTITUTE AND LOCATION 
NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 
0.27 



PROFESSIONAL: 

0.27 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



(b) Human tissues G (c) Neither 



SUMMARY OF WORK <Us9 standartt unraHucad typu. Oa not excaud the spaca pmnOad.) 

Ocular specimens from human ocular tissues with various diseases, such as 
uveitis, conjunctival and corneal diseases, and ocular metabolic genetic 
diseases were studied using iiranunoperoxidase technique as well as light and ■ 
electron microscopic evaluation. In uveitis, immunocompetent cells and 
lymphokines are critical in the reflection of clinical diagnosis, disease 
course and prognosis. In non-uveitis, alteration of cellular membrane surface 
markers and intracytoskeleton on the ocular resident cells may imply damages 
and abnormalities in these diseases . 



PHS 6040 (Rav. 1/84) 



ere •i4^i( 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

i ZOl EY 00231-03 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 ctiaractars or less. Wa must tit on one line tjetween tne oomers.) 
Cell Surface Antigens on Retinoblastoma Cells 



PRINCIPAL INVESTIGATOR (List oWer pmtassional personnel oelow the Principal Investigator) (Name, title, laboratory, ana insotuta alfiliationl 
PI: Barbara Detrick Ph.D. Expert LI, NEI 



Others: John J. Hooks Ph..D. 

Gerald J. Chader Ph.D. 
Caroline Percopo B.S. 



Head, Section on Immunology LI, NEI 

and Virology 
Chief LROffi, NEI 

Biologist LI, NEI 



COOPERATING UNITS (it any) 

Charles 



Evans 
Norman Katz 
Merlyn Rodrigues 



M.D. Head, Tumor Biology Section LB, 
M.D. Walter Reed Army Medical Center 
M.D. University of Maryland, Baltimore 



NCI 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.6 



PROFESSIONAL: 

0.4 



OTHER: 



0.2 



CHECK APPROPRIATE BOX<ES) 

dl (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



D (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Usa smnoaiti unreducad type. Do not axcaatf ma spaca promlatt.) 

Retinoblastoma (Rb) , an ocular tumor of childhood, consists of multipotent 
embryonic cells that have the potential to differentiate into neuronal or 
glial-like components. MHC class II antigens (HLA-DR, DQ, DP) are integral 
glycoproteins which are critical elements in immune regulation. The 
identification of these determinants on a variety of primitive stem cell types 
and tumor cells arrested at selected phases of their cell cycle has suggested 
that these molecules play a role in cellular differentiation. 

Recently, we demonstrated the presence of the class II molecules on Rb cells. 
In addition, the modulation of HLA-DR by IFN-y as well as the preferential 
expression of this determinant over HLA-DQ is described. Double labeling 
experiments revealed that HLA-DR antigen is shared concomitantly with cells of 
glial and neuronal character. 

Based on these initial studies, additional investigations are in progress. 
One approach focuses on the correlation of class II antigen expression with 
cellular differentiation. A second examines the prognostic significance of 
these molecules on retinoblastoma cells and the possible relationship these 
proteins may have to the modulation and management of this tumor. Finally, a 
third study will examine the role of IFN-gamma as a differentiating agent of 
this tiimor. 



PHS 6040 (Rav. 1/84) 



SPO •I4-*II 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PPIOjECT -.bMBE = 



ZOl EY 00235-03 LI 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



j TITLE OF PROJECT ISO characters or less. Title must ht on orw line Oafweon the boraers.) 

Identification and Modulation of Class II Antigens 



PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator) (Name. otte. laboratory, ana institute atfillaOon) 

PI: Barbara Detrick Ph.D. Expert LI, NEI 



Others: John J. Hooks Ph.D. 

Chi-Chao Chan M.D. 

Caroline Percopo B.S. 

Robert B. Nussenblatt M.D. 



Head, Section on Immunology LI, NEI 

and Virology 

Medical Officer LI, NEI 

Biologist LI, NEI 

Clinical Director NEI 



COOPERATING UNITS (it any) 

G. Aguirre 
Barton F. Haynes 
Laurence Boumsell 



D.D.S., P.D. Univ. of Pennsylvania 
M.D. Duke University 

M.D. Paris, France 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunology and Virology 



INSTTTUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.44 



PROFESSIONAL: 



0.34 



OTHER: 



0.1 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (al) Minors 
n (a2) Interviews 



D (b) Human tissues [1 (c) Neither 



SUMMARY OF WORK (Use ttanOanl unreduced type. Do not exceed the speee provided.) 

Class II antigens are integral glycoproteins encoded by genes in the major 
histocompatibility coit^jlex. Their expression is critical to immune reactivity. 
Although most immune cells constitutively express class II antigens, some 
non- immune cell types can be induced to demonstrate these molecules under 
selected conditions, such as an immunologic or degenerative event. Based on our 
earlier data, demonstrating that retinitis pigmentosa patients had an alteration 
in IFN-gamma production and class II antigen expression, we expanded our studies 
to evaluate class II antigen expression in a variety of ocular situations. We 
found that the retinal pigment epithelium cell does not express class II antigen 
in the normal eye. In contrast, the rpe cell did express these molecules in a 
retinal degenerative disorder (retinitis pigmentosa) and in two ocular inflam- 
matory diseases (synpathetic ophthalmia and uveitis) . Using the EAU animal 
model of ocular autoimmune disease we demonstrated that the rpe cell is acti- 
vated to express class II antigens pridr to clinical and histopathological 
evidence of the disease. Finally, we demonstrated that EAU could be altered 
with anti-la therapy. In this study EAU animals receiving monoclonal anti-la 
antibodies experience not only less ocular inflammation but also a delay in the 
onset of EAU. Moreover, immunocytochemistry analysis revealed that eyes from 
these animals expressed less la antigen as well as a diminution of infiltrating 
macrophages and lymphocytes. These data show that anti-la treatunent signifi- 
cantly modifies the course of EAU in the rat. We have also demonstrated that 
direct inoculation of recombinant IFN-gamma results in the expression of MHC 
class II (la) in a variety of ocular cells. We are continuing to investigate 
the effects of other potent modulators with the hope that an alteration in 
activation or expression of these molecules may modify the disease process to 
the benefit of the host. 



PHP pajn (Pau 1 'Ml 



SPO •I4.»1« 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT I ^^l EY 00092-10 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Vtle must lit on one line between the tiorclers.) 
HLA, ABO, and B-cell Alloantigens and Ocular Inflammatory Disease 



PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Pnnapal Investigator) (Name, title, laboratory, and institute altillation) 
PI: Robert B. Nussenblatt M.D. Clinical Director NEI 



COOPERATING UNITS (it any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immuno regulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEAHS: 
0.03 



PROFESSIONAL: 

0.03 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

G (a) Human subjects D (b) Human tissues D (c) Neither 

D (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Use standard unreduced type. Do not axcaod tfis apace provided.) 

Patients with ocular toxoplasmosis, pars planitis, Behcet's disease, 
chorioretinitis of unknown origin, are being studied to determine the phenotype 
frequency of the HLA, ABO, and B-cell alloantigens . Because the B-cell 
alloantigens or DR antigens are thought to play a role in the immunologic 
response to antigens, these findings will complement other immune uveitis 
studies being simultaneously carried out. Restriction fragment analysis has 
begun to complement these HLA studies . 



PHS 6040 (Rev. 1/84) SPO 9i4.ait 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 00075-10 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (90 characters or less. Title rrtust tit on one line between me borders.) 
Immune Functions in Ocular Diseases of Obscure Etiology 



PRINCIPAL INVESTIGATOR (List other protessiortal personnel below the Principal Investigator) (Name, title, laboratory and institute amilatkm) 
PI: Robert B. Nussenblatt M.D. Clinical Director NEI 



Others; 



Alan G. Palestine 


M.D. 


William Leake 


M.S. 


Rashid Mahdi 




Janet L. Davis 


M.D. 



Head, Section on Clinical LI, NEI 

Immunology 

Biologist LI, NEI 

Biologist LI, NEI 

Senior Staff Fellow LI, NEI 



COOPERATING UNITS (If any} 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN- YEARS: 
1.60 



PROFESSIONAL: 
0.40 



OTHER: 



1.2 



CHECK APPROPRIATE BOX(ES) 

Izl (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



D (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use sfndu^ unrvlucad type. Do not exceed the aptce provldtd.} 

In vitro cellular immune functions and lymphocyte subsets are being studied in a 
masked method in patients with ocular toxoplasmosis, pars planitis, Behcet's 
disease, geographic choroiditis, and chorioretinitis of unknown origin. Crude 
ocular antigens, purified uveitogenic soluble antigen (S-antigen) , IRBP of the 
retina, and uveitogenic fractions of the retinal S-antigen are being used in a 
lymphocyte microculture technique to evaluate the presence of cellular immune 
memory to ocular tissues. In addition, purified antigens from the toxoplasmosis 
organism are also being tested in this in vitro system. A subgroup of patients 
with posterior uveitis has been identified as having this iimnunologic memory. 
Lymphocyte subsets in the blood and in the eye are being defined in these 
patients by monoclonal antibodies which may shed light on the basic mechanisms 
of uveitis and may be used as a guide for specific immunologic therapy. The 
serum from these patients is also being evaluated. Using retinal biopsy, a new 
retinopathy in AIDS appears to have been identified. 



PHS 6040 (Rev. 1/84) 



SPO tt4.»lt 



PROJECT DUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT ^°^ ^^ 00094-10 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (30 cfiaracters or less. Title must lit on one line tsetween the txrders.) 
Immune Mechanisms in Experimental Autoinmiune Uveitis 



PRINCIPAL INVESTIGATOR (List other protessional personnel 6e/ow the Pnncipat Investigator) (Name, title, laboratory, ana institute affiliation) 

PI: Robert B. Nussenblatt M.D. Clinical Director NEI 

Others: Phuc Le Hoang M.D. Visiting Scientist LI, NEI 

Rashid Mahdi Biologist LI, NEI 



COOPERATING UNITS (if any) 



UVa/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.7 



PROFESSIONAL: 

0.6 



OTHER: 

0.1 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects G (b) Human tissues ^ (c) Neither 

n (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Usa standard unreduced typa. Do not exceed the space provided.) 

Lewis rats and non-human primates, immunized at a site distant to the eye with 
the retinal soluble antigen (S-antigen) in complete Freund's adjuvant, develop 
experimental autoimmune uveitis (EAU) . Lymph node cells and peripheral 
lymphocytes from immunized animals manifested significant cellular immune 
responses measured by the lymphocyte culturing technique. The cyclosporines, a 
family of drugs with specific anti-T-cell-activity, have been found to be 
exceptionally effective in protecting rats with EAU. Attempts at local 
immunosuppressive therapy in order to prevent EAU have begun. Topical and 
periocular cyclosporine-A (CsA) have been used in order to evaluate its 
effectiveness in EAU. Newer cyclosporines, particularly D&G, have been 
evaluated in this model, with their efficacy compared to that of CsA. Ciamexone, 
a drug with immunopotentiating characteristics, has always been utilized in this 
model. The use of "natural" immunomodulatory models are being developed. 



PHS 6040 (Rev. 1/84) gpo 9I4-«i* 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00115-08 LI 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must tit on one line Between the Ooraers.) 
Cyclosporine Therapy in Uveitis 



PRINCIPAL INVESTIGATOR (Ust ott)er professional personnel below the Principal Investigator.) (Name, title, laboratory, ana instituta affiliation) 
PI: Robert B. Nussenblatt M.D. Clinical Director 



NEI 



Others; 



Alan G. Palestine 


M.D 


Janet L. Davis 


M.D 


Jeffrey N. Bloom 


M.D 


David C. Herman 


M.D 


Chi-Chao Chan 


M.D 



COOPERATING UNITS (if any) 



Head, Section on Clinical LI, NEI 

Immunology 
Senior Staff Fellow LI, NEI 

Senior Staff Fellow LI, NEI 

Senior Staff Fellow LI, NEI 

Medical Officer LI, NEI 



UkB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 
0.75 



PROFESSIONAL; 

0.75 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

m (a) Human subjects 
n (al) Minors 
n (32) Interviews 



D (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standani unreducad type. Do not axcaad tfi* spaca prmidad.) 

Cyclosporine, an endecapeptide fungal product with specific anti-T-cell 
characteristics, will be administered to patients with sight-threatening ocular 
inflammatory disease of non-infectious origin who have failed on either 
corticosteroid or cytotoxic agent therapy. This will be done to test 
cyclosporine 's efficacy in the treatment of uveitis. Within the context of 
these ongoing studies, the effect of hydergine on reversing cyclosporine induced 
nephrotoxicity is being evaluated in a randomized, masked, cross-over study. 
Additionally, selected patients whose uveitis is well controlled on cyclosporine 
for one year or more are undergoing kidney biopsies to evaluate the long term 
effects of this agent. A phase I/II randomized trial using Cyclosporine A and G 
has begun. 



PHS 6040 (Rev. 1/84) 



SPO 9I4'4II 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

ZOl EY 00228-03 LI 



I 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (BO cfiaracwn or less, nie must tit on one line between the txjraers.) 

Study of Ocular Glial Cells Involvement in Uveitis 



PRINCIPAL INVESTIGATOR (Ust other pmlessionai personnel below the Principal Investigator.) (Name, tiae. laboratory, and institute atfiliatlon) 
PI: Francois Roberge M.D. Visiting Associate LI, NEI 



Others: Robert B. Nussenblatt M.D. 
Rachel Caspi Ph.D. 



Clinical Director 
Visiting Associate 



NEI 
LI, NEI 



COOPERATING UNITS (H any) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Immunoregulation 



INSTTTUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 

0.92 



PROFESSIONAL; 

0.92 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (a1) Minors 
n (a2) Interviews 



n (b) Human tissues Q (c) Neither 



SUMMARY OF WORK (Us9 standanl unreOuceO typu. Oo not axcood tho spac* prmtOad.) 

The work extended our ongoing study of interactions between the retinal glial 
Miiller cell and T-lymphocytes . In an in vitro co-culture system, Miiller cells 
had been shown to exert a profound inhibitory influence on the proliferation of 
T-helper cell lines through a membrane bound factor. Investigations of the 
nature of the inhibitory moiety revealed that it was sensitive to proteinase. 
Further studies showed that the expression of the factor on the surface of 
Miiller cells could be suppressed by glucocorticoids. 



PHS 6040 '""v 1 '° 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 



NOTICE OF INTRAMURAL RESEARCH PROJECT 



ZOl EY 00249-01 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 cnaracmrs or lasa. We must tit on one line oeiweon me oomers.) 
Cytokines in Human Intraocular Fluids 



PRINCIPAL INVESTIGATOR (Ust omer pmtessionm personnel Oelow tfie fnnapal Investigator. ) (Name. atia. laboratory, ana institute altillaaon) 
PI: Janet L. Davis M.D. Senior Staff Fellow LI, NEI 

Others: Robert B. Nussenblatt M.D. Clinical Director NEI 



COOPERATING UNITS r//any> g^^ Research Institute, Boston, Massachusetts (Alex £. Jalkh, 
M.D.); Eye Research Institute, Boston, Massachusetts (Charles Schepens, M.D.); 
University of Miami, Miami, Florida (Harry W. Flynn, Jr., M.D.) 



UVB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
0.32 



PROFESSIONAL; 

0.32 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects C3 (b) Human tissues G (c) Neither 

n (a1) Minors 
n (a2) Interviews 



SUMMARY OF WORK (Um standurl unreOuctO typa. Oo not exceed ttie space prrniOed.) 

Human intraocular fluids are collected during the course of surgery to 
repair retinal detachment, remove vitreous and strip membranes of 
proliferative vitreoretinopathy (PVR) , and remove vitreous and cataract from 
uveitic eyes. These fluids (ordinarily discarded) are analyzed for 
interleukin 1 (IL-1) and interleukin 2 (IL-2) activity by bioassays. 
Specimens from 88 patients have been analyzed for IL-1. Il-l activity was 
found in 90% of uveitic eyes, 35% of eyes with retinal detachment and 17% of 
. eyes with proliferative vitreoretinopathy. Thirty-two specimens have been 
analyzed for IL-2. Ten percent of uveitis patients, 20% of retinal detachment 
patients and 60% of PVR patients had detectable IL-2 activity. IL-1 is a 
mediator in multiple organ-specific pathways. Its presence in the eye 
suggests a role in intraocular inflammatory and immune processes. IL-2 is 
produced by activated T cells. The high percentage of PVR patients with IL-2 
activity suggests a role of the immune system in proliferative 
vitreoretinopathy . 



PHS 6040 (Rav. 1/84) avo*i4-*it 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00248-01 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 cfiaractars or less. Wa must m or on« line between the tioroers.j 

Magainin Therapy of Infectious Keratitis 



PRINCIPAL INVESTIGATOR (List otrier protessmnm personnel Oeiow tne Pnnap*! Investigator.) (Name, mie. letxyatory. ami msotute attilianon) 

PI: Phuc Le Hoang M.D. Visiting Scientist LI, NEI 



Others: Robert B. Nussenblatt M.D. 
Janet L. Davis rl.D. 

Rashid Mahdi 



Clinical Director 
Senior Staff Fellow 
Biologist 



NEI 
LI, NEI 
LI, NEI 



COOPERATING UNITS (It any) 

Human Genetics, National Institute of Child Health and Human Development 
(Mchael Zasloff, M.D,, Ph.D.); Human Genetics, National Institute of Child 
Health and Human Development (Charles Bevins, H.D., Ph.D.) 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 

Section on Immunoregulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

1.5 



PROFESSIONAL: 

1.0 



OTHER: 



0.5 



CHECK APPROPRIATE BOX<ES) 

n (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



D (b) Human tissues H (c) Neither 



SUMMARY OF WORK (Use stanoan unnOuoaa typa. Do not axoaea Ote space pmvtOed.) 

Studies in animals are being carried out to determine the in vivo activity of 
a new class of antimicrobial peptides isolated from the skin of the African 
frog Xenopus laevis and called magainins. This family of peptides consists of 
two closely related peptides that are each 23 amino acids which inhibit growth 
of numerous species of bacteria and fungi in vitro. An animal model of 
experimental bacterial keratitis induced in adult New Zealand white rabbits 
was used to determine the in vivo relevance of the antimicrobial activity of 
magainins. Pseudomonas aeruginosa corneal infection was primarily considered 
because it is the most destructive and the most difficult to treat corneal 
infection in humans. Each cornea was infected by an intrastromal injection of 
100 bacteria. The topical treatment with magainin drops or ocular ointment 
was started either 4 hours or 20 hours after the infection. The control 
animals were either not treated or treated with the vehicle (PBS or petrolatum 
plus mineral oil) . These preliminary studies demonstrated the in vivo 
activity of the magainin by showing a less severe corneal abscess in the 
treated animals with a delayed onset of the abscess as compared to the control 
animals. Although the animals could tolerate well the treatment, magainin 
drops and ointment induced a chemosis with a conjunctival hyperhemia by 
themselves which can aggravate the conjunctival inflammation related to the 
infection. 



PHS 6040 (Rev. 1/84) 



GPO ■l4-*lt 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 

I ZOl EY 00132-07 LI 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (30 chancters or lass. Title must tit on one una Oetween tna Ooraars.) 
ilolecular Biology of Phototransduction 



PRINCIPAL INVESTIGATOR (List otnar protassionaj personnel Oelow the Pnnapat Investigator.) (Name. Offa. laooratory. ana insotvta attillation} 

PI: Toshimichi Shinohara Ph.D. Head, Section on Molecular LI, NEI 

Biology 



Others: Masahiko Tsuda 

Benjamin Amaladoss 
Kunihiko Yamaki 
Charles Egwuagu 
Shuji Suzuki 



M.D., Ph.D. Visiting Associate 

Ph.D. Visiting Associate 

M.D., Ph.D. Visiting Associate 

Ph.D. Staff Fellow 

M.D., Ph.D. Visiting Associate 



LI, NEI 

LI, NEI 

LI, NEI 

LI, NEI 

LI, NEI 



COOPERATING UNITS (it aityt 



LAB/BRANCH 

Laboratory of Immunology 



SECTION 
Section on Molecular Biology 



INSTTTUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 

4.13 



PROFESSIONAL; 

4.13 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



D (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Uam stunoard unreOuced type. Oo not axcted ttm space pionaed.) 

The eye has a remarkable property in that it can function efficiently over a 
very wide range of illuminations from single photon to bright sun. The rod 
cells which have photosensitive rhodopsin are more sensitive to dim light and ' 
dark adapts to increase their sensitivity. However, the rod cells cease their 
sensitive phototransduction in bright light, the cone cells are in contrast 
operative in bright light . 

Rhodopsin, transducin, PDE, rhodopsin kinase and S-antigen have been known to be 
associated with the phototransduction cascade, our focus of interest is on 
rhodopsin kinase and S-antigen. In order to further understand this light 
dependent modulatory mechanism in rod outer segments, we have characterized 
S-antigen, rhodopsin kinase, calmodulin and 24K ROS specific proteins using 
recombinant DNA technologies . S-antigen had local regions of sequence homology 
with a-transducin including the putative rhodopsin binding and phosphoryl 
binding sites . 

Rhodopsin kinase is a family of proteins which have conserved features and 
similar catalytic domains among themselves. Also, calmodulin is a family of 
Ca"'"'' binding proteins and it had conserved domins too. The 24k ROS specific 
protein did not have any sequence similarity with other known proteins. Thus, 
the amino acid sequences of these proteins further substantiated the functional 
roles of these proteins in the phototransduction cascade. 



PHS 8040 (Rev. 1/84) 



SPO •I4-«I« 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT .MUMBER 

ZOl EY 00250-01 LI 



i 



PEHIOD COVEHED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 ctiancnrs or less. Title must tit on one line oetween tne ixraers.) 
Molecular Biology of Experimental Autoimmune Uveitis 



PRINCIPAL INVESTIGATOR (List otfiar prorassional oenonnel oelow tne Principal Investigator.) (Name, title. /aOoraro/y, ana instttute attiliation) 
PI: Toshimichi Shinohara Ph.D. Head, Section on LI, NEI 

Molecular Biology 



Others: Kunihiko Yamaki 
Vijay K. Singh 
Charles Egwuagu 
Tohru Abe 



M.D., Ph.D. Visiting Associate LI, NEI 

Ph.D. Visiting Associate LI, NEI 

Ph.D. Staff Fellow LI, NEI 

M.D. Visiting Associate LI, NEI 



COOPERATING UNITS (it any} 

Larry A. Donoso 



M.D., Ph.D. Wills Eye Hospital, Philadelphia, 
PA 



UVB/8RANCH 

Laboratory of Immunology 



SECTION 
Section on Molecular Biology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEAHS: 

2.2 



PROFESSIONAL: 

2.2 



OTHER: 



D (b) Human tissues G (c) Neither 



CHECK APPROPRIATE BOX(ES) 

El (a) Human subjects 
Q (al) Minors 
D (a2) Interviews 

SUMMARY OF WORK (Uaa stanOuxl uniwJucad type. Oa not excaeti ttie space provioaa.) 

Inf laitimatory diseases of the eye are a significant cause of visual handicap in 
the United States and throughout the world with severe cases often leading to 
blindness. Autoimmune processes directed against normal eye tissues, such as 
the retina, are thought to play a significant role in the pathogenesis of such 
diseases . Molecular mimicry, a process by which an immune response directed 
against a non-self protein cross reacts with a normal host protein, may play a 
role in autoimmunity. Experimental autoimmune uveitis (EAU) serves as an animal 
model of ocular inflammation. The disease is caused by the immunization of 
microgram amounts of a soluble retinal protein, designated S-antigen, in 
susceptible animal strains, including primates. 

We have determined amino acid sequences of human, mouse and bovine S-antigen. 
Immunogenic sites and four uveitopathogenic sites using 20 different chemically 
synthesized oligopeptides were also determined. We induced EAU and pinealitis 
in Lewis rats with a small synthetic peptide, corresponding to amino positions 
106 to 117 in yeast histone H3, which incidentally contains five consecutive 
amino acids identical to a uveitopathogenic site in human S-antigen. In 
addition, native yeast histone H3 was also capable of inducing an EAU. These 
findings provide a basis for autoimmune inflammatory diseases of the eye in 
humans . 



PHS 6040 (Rav. 1/84) 



SPO •I4.»lt 



ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October 1, 1987 - September 30, 1988 

REPORT OF THE CHIEF, LABORATORY OF MECHANISMS AND OCULAR 

DISEASES 
Jin H. Kinoshita, Ph.D. 



A quarter of a century ago, Drs. Cogan and Kuwabara introduced the 
hypothesis that the initiating factor in diabetic retinopathy was the 
selective loss of the retinal capillary pericytes. Drs. Kador and Akagi 
thought that the degeneration of the pericytes in diabetes was due to aldose 
reductase (AR). In order to support their hypothesis, it was essential to 
demonstrate the presence of AR in pericytes. Where others have failed, Drs. 
Kador and Akagi demonstrated immunohistochemically that AR was found in the 
pericytes and not in the endothelial cells of human retinal capillaries. More 
recently this was confirmed by others in our laboratory who showed that cell 
cultures of human retinal capillary pericytes do contain AR as demonstrated by 
biochemical, immunohistochemical , and molecular biological techniques. 

Recently Drs. Kador and associates have initiated diabetic retinopathy 
studies in galactosemic dogs. This model has been shown by Engerman to 
develop a background retinopathy which was indistinguishable from that of 
diabetic dogs. Dr. Kador and associates have been following the progression 
of retinal changes in both AR treated and untreated galactosemic dogs. They 
found that along with with pericyte ghosts in untreated dogs, there was 
proliferation of endothelial cells, the presence of acellular capillaries and 
later microaneurysm formation. All these retinal changes in galactosemic dogs 
were prevented by treatment with aldose reductase inhibitor. 

Similar results were observed in the rat model. Although rats were not 
known to develop diabetic retinopathic changes, Dr. Robison has recently shown 
retinal micro-and macrovascular changes in long term galactosemic rats. He 
also found loss of pericytes, proliferation of endothelial cells and 
microaneurysms. Different from the galactosemic dogs, were the engorged 
veins, venules, and capillaries in the retina of galactosemic rats. All these 
.retinal changes in galactosemic rats were prevented by an aldose reductase 
inhibitor. 

These studies emerging from the laboratories of Drs. Kador and Robison are 
most significant and may pave the way in the development of a new treatment 
modality for diabetic retinopathy. 

Another research area of active progress is the study on gyrate atrophy 
(GA) in Dr. Inana's laboratory. 



Gyrate atrophy is a blinding autosomal recessive degenerative disease of 
the retina and choroid of the eye characterized by a generalized deficiency of 
the mitochondrial enzyme, ornithine aminotransferase (OAT). The knowledge of 
the underlying biochemical defect in GA enabled Dr. Inana to take a molecular 
genetic approach in studying this disease. First, he constructed and 
characterized a molecular probe for the human OAT in the form of a cDNA 
clone. Analysis of the cDNA-derived OAT sequence revealed the presence of an 
OAT precursor containing a leader sequence similar to those found in other 
mitochondrial proteins of cytoplasmic origin. A differential hybridization 
analysis of the human genome using specific OAT cDNA-derived probes 
demonstrated the presence of one putative functional OAT gene and at least 
three other OAT-related genes indicating a gene family. The functional OAT 
and OAT-related gene sequences were mapped to a precise area of chromosones 10 
and X. A sequence analysis of OAT gene clones confirmed the chromosone 10 
gene to be the functional gene and at least one of the X chromosone genes to 
be a pseudogene. Analysis of the OAT gene, mRNA, and protein in 20 GA 
patients using the OAT DNA and antibody probes demonstrated a GA case with a 
partial heterozygous deletion of the OAT gene, no OAT mRNA, and undectable 
level of OAT protein. The rest of the cases showed normal OAT gene and 
variably reduced levels of OAT mRNA and protein. In one of the cases the OAT 
mRNA level was shown to be half of normal, indicating expression of only one 
of the OAT gene alleles, and a point mutation was demonstrated in the 
expressed mRNA resulting in an amino acid change in the OAT protein. The 
results from these cases constitute the first real demonstration of the 
molecular genetic defect of OAT present in GA. 



PROJECT iMUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 

ZOl EY 00245-01 LMODi 



'!5l'^ofeer"!° 1987 to September 30, 1988 



TDlEiOF PRQJECTJ80 cbaractors ot/ess. Title must fit on one line Between tne tjoraers J 
Molecular Biology or Cataracts 



PRINCIPAL INVESTIGATOR (Ust Other professional personnel Oelow the Principal Investigator.) (Name, title, laboratory, ana institute attillabon) 
PI: Teresa Borr^s Ph.D. Expert LMOD, NEI 

Others: Anna Rodokanaki M.D. Visiting Fellow LMOD, NEI 



COOPERATING UNITS (it any) 

Karolinska Institute, Stockholm, Sweden (Dr. Hans Jornvall) 
NIADDK, Diabetes Branch (Dr. Flora de Pablo) 



Mechanisms of Ocular Diseases 



SECTION. ^ „ ^ 

Section of Cataracts 



INSTTTUTE AND LOCATION , 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

1.8 



PROFESSIONAL: 

1.6 



OTHER: 

0.2 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects D (b) Human tissues \^ (c) Neither 

D (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK OJae standanl unrMucad type. Do not exceed the spaca providad.) 

Cataract development is a complex process involving a whole range of 

different causes. The hereditary cataracts in our animal model provide an 

excellent opportunity to identify changes in gene regulation that will result in 

the formation of lens opacity. The study of gene expression changes in 

hereditary cataract of strain 13/N of guinea pigs is particularly important 

because it provides the only model of nuclear hereditary cataract and because 

guinea pigs, as humans, are born with their eyes open, when this cataract is 

already present. 

A new guinea pig lens crystallin, C-crystallin, discovered in our section by 
Drs. Huang-Zigler, appears to be absent or sharply reduced in the lens of the 
cataractous animal. We set up to clone the copy of the mRNA encoding Zeta- 
crystallin as a first approach to understand which step of the regulation of 
this gene could be responsible for the defvelopment of the cataract. 

We screened a guinea pig cDNA library with a synthetic oligonucleotide and 
obtained a positive clone (pTBlOO) of 1448 base pairs (bp). This clone contains 
the full C -protein coding region of 328 amino acids with a MW of 35,071 daltons; 
it provided us with the first primary structure of this novel protein. 

Computer search of the ^-crystallin sequence in the protein sequence data 
bank revealed a 33% similarity of this protein with the alcohol dehydrogenase 
' (ADH) protein family. Further analysis of the comparison proved to be 
statistically significant indicating that C -crystallin belongs to the 
superfamily of the ADHs. The role of this similarity and its significance in 
lens transparency is under present study. 



PHS 6040 (Rev. 1 /84) a PO 9 1 4-a i ■ 



DEPARTMENT Of HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00201-04 LMOD 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (BO cnamemra or wss. Tnm muti M on on* «n* ourwaan 

Molecular Biology of Aldose Reductase 



fns 0ofO9n.t 



PRINCIPAL INVESTIGATOR (Lai otntr 

PI : Deborah Carper 

Others: Chihiro Nishimura 
Caroline Graham 
Masayuki Kaneko 



1/ oatow ma f>nnaomi immagator.) (Namt. nM. MCoranyy. ana twaum artuianoni 

Ph.D. Biologist LMOD, NEI 

M.D. Visiting Associate LMOD, NEI 

B.A. Chemist LMOD, NEI 

M.D. Visiting Associate LMOD, NEI 



COOPERATING UNITS III any) 

David J. Barrett, Jules Stein Eye Institute, UCLA, Los Angeles, CA 



LAB/BRA NCh 

• Laboratory of Mechanisms of Ocular Diseases 



SECTION 
Section on Cataracts 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN. YEARS: 



PROFESSIONAL: 



3.0 



3.0 



OTHER: 



0.0 



CHECK APOROPRIATH aOX/ESi 

C (a) Human suojects 
C (al) Minors 
d (a2) Intervievys 



C (b) Human tissues iZi (c) Neither 



SUMMARY Of WORK (Um sanoBro umwtkjemt lyw. Oo not mcmo ma cote* ofOMoaa; 



Aldose reductase (AR) of the polyol pathway has been in^Jlicated in some of 
the disabling complications of diabetes . We have now successfully completed the 
protein sequence for aldose reductase using cDNA sequencing and primer extension 
analysis of AR mRNA. The. primary structure of AR consists of an open reading 
frame of 948 nucleotides encoding for a 316 amino acid polypeptide (including 
the initiation methionine) with a molecular weight of 35,797. Secondary 
structure predictions indicate that AR is over 50% p-Sheet. 

Protein comparisons have previously revealed structural relatedness (41% to 
57%) among vertebrate aldose reductase, aldehyde reductase, prostaglandin F 
synthase and the frog lens protein p-crystallln. This superfamily can now be 
extended to prokaryotes by the inclusion of Corynebacterium 2,5 diketo-D- 
gluconate reductase. This more distantly related protein shares 30-40% identity 
with the vertebrate enzymes. 

Southern blot analysis indicated the existence of a multi-gene family for 
AR. Since our amino acid sequence data for AR have revealed considerable 
sequence similarity to other aldo/keto reductases, it will be interesting to 
elucidate the relationship between genes encoding these proteins and a gene 
family for AR. 



PROJECT NUMBER 
DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 



NOTICE OF INTRAMURAL RESEARCH PROJECT 



PEBIOD COVERED 

October 1, 1987 to September 30, 1988 



ZQl SY 00189-05 LMOD 



TITLE OF PROJECT !B0 cnancmrt or mu. Tnm mutt In on on* an* oaiwtan mm ooraon.) 

Oxidation of Proteins in Cataractogenesis 



PRINCIPAL INVESTIGATOR (ust om»r orowsstona/ otnomm ovow me Pnnatfi irvtmsngator.) lNam» tm*. Mooraiofy. tna nwrura attmaoon) 

PI: Donita L. Garland, Ph.D. Research Chemist LMOD, NEI 



COOPERATING UNITS it any) 

None 



LAB/BRA NCh 

Laboratory of Mechanisms of Ocular Disease 



SECTION 

Cataracts 



INSTITUTE AND LOCATTON 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-VEAaS; 

1.0 



PROFESSIONAL: i OTHER; 

1.0 ! 0.0 



CHECK APOROPRiATc BOX/ES. 

C (a) Human suojects [2 (b) Human tissues G (c) Neither 

C (al) Minors 
ZI (a2) Interviews 



SUMMARV OF WORK (Um um t j a iu mrvouoae tyoa. Oo net axeama tna aoaoa pmioaa.) 



Oxidative changes of lens proteins are thought to occur with aging and to 
contribute to the development of cataracts. The goals of this project are to 
determine: 1) the extent of oxidative modification of crystallins and metabolic 
enzymes in both normal and cataractous lenses; 2) the nature of the modifications 
and mechanisms leading to the changes; and 3) the effect of the modifications on 
structure and function of lens proteins. Bovine and human lenses were used. The 
approach has been to study the modifications of lens proteins after treatment in 
vitro by mixed function oxidation systems. Treatment of bovine Ya'crystallin 
caused the loss of about two sulfhydryls and a progressive loss of methionine 
residues with increased time of oxidation. Only a fraction of a cysteic acid 
residue was found and the modification of other amino acids has not yet been 
correlated with new species formed upon oxidation. Deamidation has yet to be 
examined. Similar studies are in progress on a human gamma crystallin expressed 
in mouse L cells; the goal is to identify the modified amino acids. The proteins 
of bovine trabecular meshwork extracted by various procedures were analyzed by 
polyacrylamide gel electrophoresis. The profile was very similar to that of 
human trabecular meshwork. There were a few significant differences between calf 
and cow trabecular meshworks. These results suggest that bovine trabecular 
meshwork may be a useful model system to study glaucoma. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



Pf^OjECT NUMBER 



201 EY 00237-03 LMOD 



PEWOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT f80 cnamcnrs or mu. Titm mutt m on on* an* 

Characterization of the Lens 



PRINCIPAL INVESTIGATOR (Usi otnv oro/*«iionai ofton m i ovow mt f>nnatMi mvtmgator ) (N»rm. nrw. wooraiory, ana ntmim athnanoni 

PI: Paul Russell Ph.D. Research Chemist LMOD, NEI 
Others: Masao Nakamura M.D. Visiting Associate LMOD, NEI 



COOPERATING UNITS lit any) 

Division of Cancer Research, University of Toronto (S. Meakin, M. Breitman, L.-C. 
Tsui) Howe Laboratory and Harvard University (D.L. Epstein); Lab of Retinal Cell 
and Molecular Biology, NEI (S. Gentleman) 



LAB/B Ranch 
Laboratory of Mechanisms of Ocular Diseases 



SECTION 

Section on Cataract 



iNsrrruTE and location 
NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-VEASS; 



PROFESSIONAL; 



1.6 



1.6 



OTHER; 



0.0 



CHECK APWTOPHlATc BOX/ESi 

C (a) Human suojects 
G (al) Minors 
!Z (a2) Interviews 



[^ (b) Human tissues u (c) Neither 



SUMMARY OP WORK (U» 



I ifwwauoaa lyet. Oo ner axeaaa ma j 



The processes of aging in the hiunan lens have been difficult to study 
because the mechanisms by which alterations occur in the lens are not known. 
The proteins in the lens undergo distinctive changes in their charge but the 
cause of these modifications and the relationship between these alterations 
and cataract formation is not established. One way of investigating these 
changes is to study the individual proteins in vitro and deteirmine how 
modifications affect the structure and interactions of these crystallin 
proteins with other proteins. One of the major groups of proteins in the 
lens is y-crystallin. One of the ycrystallin genes has been stably 
integrated into mouse L-cells. By using the Y-crystallin expressed in the 
mouse cells, studies of the alteration of the hiiman protein in an oxidation 
system have been done. The microheterogeneity and the shift of the protein 
to more acidic forms that are observed in the aging human lens have been 
observed with the crystallin in vitro. It would appear that the many of the 
alterations that are seen in the ycrystallin in the nucleus of the human 
lens can be mimicked with a mixed function oxidation system on isolated 
proteins. Thus, many of the changes that have been reported on aging in the 
human lens may be the result of oxidative damage to the conponents of the 
lens. 

Additionally, work has progressed on the calcium binding proteins of the 
lens. These proteins, called annexins, may play a major role in development 
and differentiation in the lens. At least two of the major calcium binding 
proteins in the lens have been shown to be glycosylated. The addition of 
sugar residues on these proteins may indicate there is another level of 
control which the cell has for these very important proteins. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00105-09 LMOD 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 enaracmrz or mu. Tim mutt hi on on» an* oanttan mm oofofs.i -^ 

Structure and Composition of Lens Crystaliins with Respect to Cataractogenesis 



PRINCIPAL INVESTIGATOR (Usi omv onftvonmi ofmnmi owow ma Pnnaoai immsogator) (Namt mm. moonmry. ana ntmuta amiianon, 

PI: J. Samuel Zigler, Jr. Ph.D. Research Biologist LMOD, NEI 

Others: Qing-ling Huang M.D. Visiting Fellow LMOD, NEI 

Xinyu Du M.D. Visiting Fellow LMOD, NEI 



COOPERATING UNITS « any) ■ 

Department of Chemistry, Adelphi University (F. Bettelheim); Department of 

Ophthalmology, University of Tennessee (H.M. Jernigan, Jr.); Oakland University, 
Rochester, MI (V.N. Reddy), Alcon Laboratories (M.Lou) 



LAB/BRA NCm 

Laboratory of Mechanisms of Ocular Diseases 



! SECTION 

Section on Cataract 



iNsrrruTE and location 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAIK-VEAHS: i PROf=ESaONAL. 



2.2 I 2.2 0.0 



OTHER: 



CHECK APOROPRIATc 90X/ESi 

C (a) Human suojects \2- (b) Human tissues u (c) Neither 

C (a1) Minors 
'~ (a2) Intervjews 



SUMMARY Of WORK (Um Mtmiomia unwauoaa ryot. Oo net axeaae mm toaem oromama.) 

Lens crystaliins are evolutionarily conservative proteins that are the 
primary structural constituents of the lens. The focus of work in this 
laboratory is oriented toward: 1) increased understanding of the structural 
attributes of these proteins which contribute to their fitness to serve as 
components of a transparent tissue and 2) elucidation of the mechanisms whereby 
changes in the composition of lens crystaliins or aging-related modification of 
these long-lived proteins can contribute to opacification of the lens. 

The studies on zeta-crystallin, a lens protein, thus far found only in guinea 
pigs, have yielded several significant new findings. We now know that this 
protein is related to alcohol dehydrogenase and thus apparently represents the 
first reported example of a taxon-specif ic crystallin in a mammal in which an 
enzyme has been adopted by the lens as a structural protein. Since zeta- 
crystallin is not present in the animals homozygous for the congenital cataract 
trait, it is possible that the lack of zeta may be the initiating factor in the 
formation of the cataract. Such a situation would provide a unique system for 
studying the function of an individual crystallin as part of the transparent 
protein matrix in the lens. Studies on protein synthesis in the cataract lenses 
reveal significant synthesis of a protein which is not detected in normal lenses. 
Use of an antibody raised against a synthetic peptide from zeta-crystallin reveals 
that this second protein is related to zeta. 

It has been demonstrated that both inhibition of the glutathione redox cycle 
with BCNU or decreasing lens ATP through use of 2-deoxyglucose can potentiate the 
oxidative modification of crystaliins in cultured rat lenses ejq>osed to hydrogen 
peroxide . 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJEC MUMBER 



I 



ZOl EY 00193-05 LMOD j 



: PERIOD COVERED 

i October 1, 1987 to September 30, 1988 



I TITLE OF PROJECT (80 cnaraaers or »ss Tirie must In on one una osiwsen me ooraan.) 

I Molecular Biology of Hereditary Eye Diseases 



PRINCIPAL INVESTIGATOR <Usi omer oroiassionai oertonrmi oeicm me frmaoai momatigator I (Name mie. mooramrv. ana msmuie aitmanoni 

PI: George Inana M.D., Ph.D. Section Head LMOD, NET 

Dthers: Carmelann Zlntz Ph.D. Staff Fellow LMOD, NEI 

Yoshihiro Hotta M.D. Visiting Associate LMOD, NEI 

Carolynn Chambers Ph.D. IRTA Fellow LMOD, NEI 

Tetsuo Sasabe M.D. , Ph.D. Visiting Associate LMOD, NEI 



Keiko Fu j iki 



Ph.D. 



Professional Consultant LMOD, NEI 



I COOPERATING UNITS (it any) 

See next page 



CHSCK ABOROORIATH aOX/sSi 

m (a) Human suoiectt 
Z (ai) Minors 
_ (a2) Imervtews 



S (b) Human tissues :_ (c) Neither 



; lAB/BHAnCm 






' Laboratory of Mechanisms of Ocular Diseases 






, SECTION 






Molecular Pathology Section 




i 


INSTrrUTk ANU LOCATION 






NEI, NIK, Bethesda, MD 20892 






TOTAL MAN-YEARS. ■ PSOFSSSONAL. 


j OTHER; 




5.4 ' 5.4 


1 






I Summary op work fU» hikjil, unrweueaa iya>. ^ nor 

Ornithine Aminotransferase Deficiency in Gyrate Atrophy: Gyrate atrophy (GA) 
Ls a blinding, autosomal recessive degenerative disease of the retina and choroid 
3f the eye characterized by a generalized deficiency in the mitochondrial enzyme, 
ornithine aminotransferase ( OAT ) . Our molecular genetic investigation of this 
disease has resulted in the cloning and characterization of a cDNA for the human 
DAT, mapping of the OAT gene sequences to chromosomes 10 and X, identification of ^ 
the OAT gene family and characterization of the members of the family including 
the functional OAT gene, construction of expression clones of OAT and expression 
Df OAT in heterologous tissues, and analysis of the OAT gene and its expression in 
3A patients which has revealed a case with a partial heterozygous deletion of the 
DAT gene and complete absence of the OAT mRNA. By examining the family members of 
this GA patient we were able to demonstrate the stable autosomal recessive 
inheritance of the OAT gene and expression defect in tJie family in addition to 
demonstrating the co-dominant mode of action of the OAT gene. Analysis of a GA 
patient who shows a marked decrease in the level of cellular OAT protein revealed 
that he is expressing only one of the two alleles of the OAT gene and that the 
expressed OAT contains a single point mutation resulting in an amino acid change. 
This amino acid change appears to modify an a-helical region of the OAT protein, 
and assay of the mutant OAT protein for mitochondrial transport/processing seems to 
indicate that the mutant protein fails to become processed. 

Hereditary Retinoblastoma and X-linked Ocular Diseases: Work on hereditary 
jretinoblastoma is continuing with isolation of malignant revertants of non- 
nalignant hybrids between Y79 retinoblastoma and NIH3T3 cells and expression 
cloning of genes that may alter the phenotype of retinoblastoma. A linkage of the 
DAT-related X chromosome genes to Norrie Disease and X-linked retinitis pigmentosa 
has been established using restriction fragment length polymorphisms detected by 
the OAT probe. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PERIOD COVERED 

October, 1987 to September, 1988 



PROJECT NUMBER 



SOI EY 00003-16 LMOD 



TITLE OF PROJECT (80 characters or less. Title rr^ust fit on one line between the borders.) 

Pharmacology of Ocular Complications 



PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, ana institute affiliation) 

PI: Peter F. Kador Ph.D. Research Chemist LMOD NEI 



Others: Laure Caspers-Velu M.D. 
Hitoshi Ikebe M.D. 
Toshihiro Nakayama Ph.D. 
Sanai Sato M.D. 
Susan DiCamillo B.A. 



Visiting Scientist 
Visiting Scientist 
Visiting Scientist 
Visiting Scientist 
Guest Worker 



LMOD NEI 

LMOD NEI 

LMOD NEI 

LMOD NEI 



COOPERATING UNITS (it any) 

None 



LAB/BRANCH 

Laboratory of Mechanisms of Ocular Disease 

SECTION 

Section of Molecular Pharmacology 



INSTITUTE AND LOCATION 



National Eye Institute. National Institutes of Health, Bethesda. Maryland 20892 



TOTAL MAN-YEARS: 
6 



PROFESSIONAL: 

5. 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects D (b) Human tissues ?II (c) Neither 

D (a1) Minors 
n (a2) Interviews 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 

The onset and progression of yarious ocular complications are being investigated 
as well as methods for their potential pharmacological control. Specifically, 
relationships between diabetes and galactosemia-induced retinopathy, cataract, 
keratopathy, and changes in pupil function, iris and ciliary process structure 
and the interactions of the enzymes aldose reductase and aldehyde reductase are 
being investigated. Methods for either delaying or preventing the onset and 
.progression of these complications through the pharmacological control of these 
enzymes are also being developed. 

Events leading to the formation of several types of cataracts are also being 
studied as well as methods for controlling the onset of these cataracts through 
pharmacological intervention. 



PHS 6040 (Rev. 1/84) 



DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



P€HIOD COVERED 

October 1, 1987 to September 30, 1988 



PROJECT NUMBER 



7.ni T7.y nn 743-Q2 T.Mon 



TITLE OF PROJECT (80 criMraenn or wu. Titm mutt tn on oim mm mmmh rrw ooroarz.) 
Ocular Cells Cultured Under Normal and Diabetic Conditions 



PRINCIPAL INVESTIGATOR (Uai omw ^rDr»s«ion«< otnommi tmow me Pnnao,! mymsogttof , (N,m, nrw. moormnry. tna ^^^mut, titmnon, 

PI: Bruce A. Pfeffer Ph.D. Senior Staff Fellow LMOD, NEI 
Others: W. Gerald Robison Ph.D. Chief, Section on 

Pathophysiology LMOD, NEI 



L 



COOPERATING UNITS « any) 



None 



I LA8/BRANCH 

' Laboratory of Mechanisms of Ocular Diseases 
SECTION ■ ' 



Section on Pathophysiology 



INSTTTUTE AND LOCATION 

NEI, NIH, Bethesda, MD 20892 



TOTAL MAN-YEARS: 

1.0 



PROFESSJONAL. I OTHER; 

2.0 I 0.0 



CHECK APPROPRIATE M»ESi ^^^^^^^^~~^~~^~" 

G (a) Human suojects £2 (b) Human tissues C (c) Neither 

C (ai) Minors 
Zl (a2) Imerviews 



SUMMARY Of WORK lUm tanmro iMwmauema tytm. Oo nerncMa m* sok* oro«<OM. 



Along with the vascular lesions characteristic of diabetic retinopathy, 
considerable clinical evidence exists that the retinal pigment epithelium (RPE) 
is affected in diabetic eye disease. Biochemical and physiological studies of 
animal models suggest that diabetic pigment epitheliopathy may be a complication 
mediated by the activity of the enzyme aldose reductase. We are utilizing 
cultured human and monkey RPE as an in vitro model system to study the effects of 
elevated hexoses on these cells. In common with other tissues in the presence of 
high sugar concentrations, transport of the amino acid taurine into cultured RPE 
cells incubated with galactose is impaired. In addition, the galactose- treated 
cells are "leakier" in such a way as to actually extrude taurine. Both of these 
effects can be partially prevented by inciibation with aldose reductase inhibitor 
(ARI) supplemental to the galactose. Since taurine is essential for normal 
retinal function, a deficit in RPE handling of taurine under diabetic conditions 
may contribute to retinal pathology. 



DEPARTMENT Of HEALTH AND HUMAN SERVICES • PUBLIC HEALTH SERVICE 



I NOTICE OF INTRAMURAL RESEARCH PROJECT 

I 



ZOl EY 00149-15 LMOD 



1 PERIOD COVEHEO 






Ontnbfir 1. T^R7 tn Rflptfimhfir ID. 1 qRR 






TITLE OF PROJECT (BO cnarwemn or mu Jim imtti ir on ont mt omiw9un am ooratn i 






Ultrastructure and F^inction nf t-.hfi CrI 1 s flnri Ti=^P;nPP; of ths F.vfi 






PRINCIPAL INVESTIGATOR <Uji MTwr orefMSonai otrtonnti emnm irw frwtenmi nmtifio' i INamt mm moortmry ana nttnun Ulutmiioni 




PI: W. Gerald Robison, Jr. Ph.D. Chief, Section on 


LMOD, 


NEI 


Pathophysiology 






Others: Masao Nagata Ph.D. Visiting Associate 


LMOD, 


NEI 


M.D. 






Bruce A. Pfeffer Ph.D. Senior Staff Fellow 


LMOD, 


NEI 


COOPERATING UNITS « any, 




1 


None 







I LAB/BRANCn 

I Laboratory of Mechanisms of Ocular Diseases 



SECTION 

Section on Pathophysiology 



INST7TUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN. YEARS i PROPSSSONA^ j OTHER 

5.2 ! 5.0 ' 



CHECK APORO»RlATc SOX/ESi 

C (a) Human suoiects E (b) Human tissues C (c) Neither 

C (31) Minors 
Z (a2) Imerviews 



SUMMARY OF WORK mm tanaan tnwoueta rftm. Oa not i 



Diabetic retinopathy is mainly a vascular disease which first manifests 

itself by several histopathological lesions related to the integrity of capillary ' 

walls, including basement membrane thickening, loss of mural cells, and ' 

endothelial cell proliferation. We now find that there is another diabetes- : 
related alteration in capillary walls which results in fewer mural-to-endothelial i 

cell contacts, and may cause endothelial cell proliferation. Normally, ! 

junctional regions which permit cell-membrane-to-cell-membrane contacts between ■ 

mural and endothelial cells occur frequently. They appear as fenestrae in the I 
thick basement membranes which separate the plasma membranes of the mural and 

endothelial cells over most of their juxtaposed surfaces. In galactose-fed rats , 

there is a significant decrease in the number of junctional regions. After 28 \ 

months of normal diet there was a mean of 1.0 (range 1-6) junctional region per i 

ultrathin transection of rat retinal capillaries, whereas, rats fed 50% | 

galactose had less than half as many (mean = 0.3). When an aldose reductase j 

inhibitor was added to the galactose diet the number of junctional regions i 

approached normal (mean = 0.8). Therefore, as with several other diabetic ' 
complications, the decrease in cell-to-cell contacts in capillary walls is 

prevented by inhibition of aldose reductase activity. The mechanism of cell ' 
contact loss will be investigated using cell culture. Aldose reductase 

inhibitors are becoming increasingly useful in studies related to the possible i 

prevention of diabetic retinopathy. ' 



P"S 60C ■=«•■. "(U 



c»c • ' «-• • • 



ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October 1, 1987 to September 30, 1988 

REPORT OF THE CHIEF, LABORATORY OF MOLECULAR AND 
DEVELOPMENTAL BIOLOGY 

Joram Piatigorsky, Ph.D. 

This is the seventh year for the Laboratory of Molecular and 
Developmental Biology (LMDB) . The efforts of this laboratory continue to be 
directed towards understanding the molecular and cellular basis for lens 
development. The complexion of the laboratory has changed this year, in that 
Dr. Toshimichi Shinohara has left to create a molecular biology section in the 
Laboratory of Immunology. We are fortunate, however, that Dr. Ana Chepelinsky 
has become a tenured member of the LMDB and has begun to form a productive 
research team. She is continuing her studies on the tissue-specific 
expression of the mouse aA-crystallin gene as she plans new investigations 
concerning the expression of non-crystallin genes in the lens. She has been a 
pivotal force in the creation of our transgenic mouse facility, presently 
housed in building 14. With Eric Wawrousek she has been able to demonstrate 
that the promoter for the mouse aA-crystallin gene retains its lens-specific 
activity in transgenic mice when a sequence containing only nucleotides -88 to 
+46 are used. This is of great interest, because in comparison with numerous 
other results we have obtained using chicken and mice it indicates that 
homologous crystallin genes do not use the same regulatory elements for their 
expression. Our ability to produce transgenic mice is continually being 
strenghtened and we now have trained two more investigators to do this 
demanding technique- -Teresa Lomjoco and Joan McDermott. 

As we continue to identify regulatory regions of the crystallin genes, 
we have increased our efforts to isolate the factors with which they interact. 
Two approaches are being developed in this connection. First, we are 
searching for lens nuclear proteins that bind to the crystallin gene 
regulatory regions. This requires the preparation and fractionation of 
proteins from lens cell nuclei. At the time of writing, David Donovan has 
resolved by ion- exchange chromatography chicken lens nuclear proteins which 
bind to the aA-crystallin promoters of chicken and mice. Working with 
Christina Sax and John Klement, Dr. Donovan has obtained preliminary evidence 
that these homologous crystallin promoters do indeed bind to different 
proteins, as the results above suggested. Moreover, it begins to look as if 
we might be able to purify these different putative regulatory proteins and 
their cDNAs. This would constitute a significant advance in our ability to 
understand how the complex temporal and spatial patterns of crystallin gene 
expression are regulated. 

Binding alone is insufficient to reconstruct the dynamics of crystallin 
gene expression. It is necessary to develop functional assays for regulatory 
factors. Dr. Sax has explored the possibility of injecting crystallin 
promoters into Xenopus oocytes as a functional test for activity. Preliminary 
results indicate that this system may be used for identifying crystallin 
transcription factors. Ultimately we may have to devise a cell-free system as 
well which behaves with specificity with respect to crystallin transcription. 



In addition to investigating the oA-crystallin gene in greater detail, 
we are also examining other crystallin genes. John Roth is in the process of 
mapping the different regulatory regions of the chicken /9B1- crystallin gene 
and George Thomas is investigating the two chicken 5 -crystallin genes. We 
have obtained evidence by using deletion mutants that these genes have 
upstream sequences repressing gene expression as well as more proximal 
positive regulatory regions. Chicken 5 -crystallin is particularly intriguing 
in that there are two extremely similar genes lying side by side on the 
chromosome, yet one is expressed about a hundred times more strongly in the 
lens than the other. It appears as if a variety of regulatory mechanisms 
govern crystallin gene expression and the challenges before us are to 
understand how any one of these operates and how the different mechanisms are 
coordinated to achieve the perfection of a transparent lens . 

Last year we reported that many crystallins, surprisingly, were 
recruited from metabolic enzymes. Graeme Wistow discovered that e -crystallin 
is similar to lactate dehydrogenase B and even has enzyme activity. We went 
on to link r -crystallin with enolase, 5 -crystallin with argininosuccinate 
lyase and the squid crystallin with glutathione S- transferase. This year Dr. 
Wistow, Tom Lietman, Barbara Norman, and I have demonstrated that these 
crystallins are encoded by the same gene as their respective enzymes, a 
situation we call gene sharing. This has important implications for the 
evolution and expression of these crystallins. From an evolutionary 
viewpoiont, gene sharing means that a single protein is under at least two 
entirely separate selective pressures, which would slow the evolutionary 
clock. It also means that the different uses of this gene, i.e. as a 
structural crystallin protein in the lens or as an enzyme in other tissues, 
evolved by modification of gene regulation alone and did not involve changes 
in the coding regions of the genes. From an expression viewpoint, gene 
sharing means that crystallins are not lens -specific, but are only 
preferentially expressed in that tissue. When the crystallin/enzyme gene is 
being utilized as an enzyme it is expressed at low levels in many different 
tissues. We must still find out whether the same or different regulatory 
sequences are used for lens and non-lens expression of a shared gene and 
whether different transcription factors are invoked when the gene is used in 
one capacity or another. The surprising and fascinating finding that 
crystallins and metabolic enzymes share genes changes our thinking of the 
evolution and regulation of crystallins. 

In contrast to the lens-specific expression of the oA-crystallin gene, 
studies by Robert Dubin have shown that the oB-crystallin gene is expressed in 
a number of different tissues, including heart, kidney and skeletal muscle. 
This suggests strongly that even crystallins with no known enzymatic function 
have another use in different tissues. Dr. Dubin showed by creating 
transgenic mice carrying an aB-crystallin minigene that lens and non-lens 
expression of this gene is regulated by its flanking sequences, most probably 
at the 5' end. One wonders how many other proteins have multifunctional 
roles, and what were the rules to select such a smorgasbord of proteins to be 
used as lens crystallins. 



In order to gain a fuller appreciation for the variety of proteins used 
as crystallins and to explore new terrain that may provide insight to the 
evolution of crystallin gene regulation, we are including invertebrates in our 
research. Last year we introduced the jellyfish as a subject of investigation 
since they have cellular lenses with a striking resemblance to vertebrate 
lenses, yet are extremely primitive animals (at least 600 million years old) 
which are, of course, built on. an entirely different body plan than 
vertebrates. We have now shown that cubomedusan jellyfish lenses contain only 
2 or 3 crystallins (depending upon species) which bear little if any 
similarity with the cirystallins of the squid or vertebrates. We have 
generated an antibody to one of the jellyfish crystallins and are ready to 
isolate its cDNA and gene. We hope that these studies into the unchartered 
waters of invertebrate crystallins will yield surprises and valuable 
information concerning the evolution and expression of these gene families. 

The work of Peggy Zelenka and her group concerns the expression of 
proto- oncogenes during differentiation of lens epithelial cells into lens 
fiber cells. Earlier studies from this section established that c-myc mRNA 
levels in cultured embryonic chicken lens epithelial explants were elevated as 
the cells withdrew from the cell cycle during differentiation. Using a 
modified nuclear run-on transcription assay which they developed, Dr. Zelenka 
has now demonstrated that the increased mRNA levels are at least partly 
regulated by increased transcription of exons 2 and 3 of the c-myc gene. Luke 
Pallansch has further demonstrated that c-myc mRNA levels in the cultured 
explants can be post- transcriptionally regulated by pharmacological agents 
which block the lipoxygenase pathway of arachidonic acid metabolism. In 
addition. Dr. Pallansch has shown that a lipoxygenase pathway metabolite of 
arachidonic acid is lost during in vitro differentiation, raising the 
possibility that this post- transcriptional mechanism may also be involved in 
the accumulation of c-myc mRNA that accompanies differentiation. 

Efforts to measure levels of c-myc protein in the past had been 
fruitless because of the failure of chicken c-myc protein to cross -react 
immunologically with available antisera against the human and mouse proteins. 
This year Howard Beswick and John Talian planned and oversaw the synthesis and 
purification of chicken-specific c-myc peptides, which were then used to raise 
antibodies in rabbits. As a result it is now possible to establish the 
relationship between c-myc protein and mRNA levels in differentiating lens 
cells. This antiserum also makes possible a variety of experiments on the 
distribution, stability, and function of c-myc protein during differentiation. 

Since expression of high levels of c-myc protein is not correlated with 
DNA replication in differentiating lens cells, other possible fxmctions for 
this protein are being considered. Dr. Zelenka, working with Anita Dash, a 
summer student, has demonstrated that elevated c-myc expression is correlated 
with accximulation of mRNA for the heat shock protein, HSP 70. In addition. 
Dr. Howard Beswick has constructed a plasmid containing a chicken c-myc cDNA 
which will allow experiments to study the effect of c-myc expression on the 
transcription of other genes in transfected cells. 



Dr. Talian has begun an investigation of the expression of a cytoplasmic 
proto- oncogene, c-src, in embryonic chicken lenses. As a preliminary step, he 
has studied the expression and distribution of calpactin I, a protein which is 
a known substrate for the tyrosine kinase activity of v-src, and which has 
recently been shown to be a major component of the lens membrane EDTA- 
extractable protein fraction by a collaborative effort between this laboratory 
and Dr. Paul Russell (NEI.LMOD). Using immunofluorescence. Dr. Talian has 
established that calpactin I has the expected localization along membranes of 
lens fiber cells, and has shown for the first time that this protein is 
present in lens epithelial cells. Using cultured explants of embryonic 
chicken lens epithelia he has shown that the intensity of immunofluorescence 
for calpactin I increases during the first 24 hr of differentiation in vitro, 
in parallel with acctimulation of calpactin I mRNA. These studies point to 
increased synthesis and accumulation of calpactin I during early stages of 
lens fiber formation, and are consistent with the suggestion that this protein 
may play a role in the cell elongation that accompanies differentiation. 

It is too often taken for granted that a laboratory runs smoothly 
without the realization that this only occurs when its support staff is 
excellent. Our secretary, Mrs. Dawn Chicchirichi, continues to take care of 
all our administrative and typing needs and we are very lucky to have her with 
us. We also rely heavily on Mrs. Barbara Norman who keeps the laboratory 
well-oiled and in top shape as she performs her "bench work". I take this 
opportunity to thank them and make my appreciation known. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00127-12 LMDB 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must fit on one line betweer^ the borders.) 

Plasma Membrane Composition and Biosynthesis in Chick Lens Fibers and Epithelia 

PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator) (Name, title, laboratory, and institute affiliation) 



PI: 



Peggy Zelenka 



Others : Luke Pallansch 
John Talian 



Ph.D. 


Geneticist 


LMDB, 


NEI 


Ph.D. 


Staff Fellow 


LMDB, 


NET 


Ph.D. 


IRTA Fellow 


LMDB, 


NEI 



COOPERATING UNITS (if any) 

Flora de Pablo 



M.D. 



Diabetes Branch, NIDDK 



LJ\B/BRANCH 

Laboratory of Molecular and Developmental Biology 



SECTION 

Section on Cellular Differentiation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 

TOTAL MAN-YEARS: 

2.5 



PROFESSIONAL: 



2.5 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects 
n (a1) Minors 
n (a2) Interviews 



D (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 



This project has identified a wide range of alterations in plasma membrane 
lipids and proteins which are associated with differentiation of lens 
epithelial cells to form lens fibers. Results have shown that 
phosphatidylinositol degradation ceases when lens epithelial cells 
differentiate to form lens fiber cells, while synthesis of phosphatidylinositol 
and other phospholipids increases. Since phosphatidylinositol is rich in 
arachidonic acid, a precursor of prostaglandins and leukotrienes , the 
metabolites of arachidonic acid produced by lens cells have been characterized. 
Loss of a lipoxygenase pathway metabolite has been correlated with the 
initiation of differentiation in vitro. Plasma membrane proteins which have 
been investigated include the insulin and IGF receptors and the membrane 
associated protein, calpactin I. Equilibrium binding studies have shown that 
embryonic chick lens epithelial cells possess both insulin and IGF I receptors , 
and that expression of both is regulated during differentiation and 
development. Analysis of membrane associated proteins has demonstrated that 
calpactin I is a major component of the EDTA-extrac table protein of lens 
membranes. mRNA for this protein accumulates during in vitro differentiation, 
in parallel with an increase in immunofluorescence staining intensity. These 
studies have shown that embryonic chicken lens epithelial membranes are dynamic 
entities which undergo structural and functional changes as part of the 
differentiation process. 



PHS 6040 (Rev. 1/84) 



GPO SI4-SIS 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00238-03 LMDB 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (SO characters or less. Title must tit on one line between the boraers.) 

Proto- oncogene Expression During Lens Differentiation and Development 



PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation) 

PI: Peggy Zelenka Ph.D. Geneticist LMDB, NET 



Others : Luke Pallansch 
Howard Beswick 
Xiu-An Zhu 



Ph.D. Staff Fellow LMDB, NET 

Ph.D. Visiting Fellow LMDB, NEI 
M.D. Visiting Scientist LMDB, NEI 



COOPERATING UNITS (if any) 



LAB/BRANCH 

Laboratory of Molecular and Developmental RioTogy 



SECTION 

Section on Cellular Differentiation 



INSTITUTE AND LOCATION 

NEI. NIH. Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

2.0 



PROFESSIONAL: 



9.0 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 



This project investigates the expression of proto-oncogenes during the 
differentiation of embryonic lens epithelial cells to form lens fiber cells, 
and seeks to determine the specific function of the corresponding gene 
products in the developing lens. Measurements of steady-state mRNA levels and 
nuclear run-on transcription experiments have identified several proto - 
.oncogenes which are actively expressed in the embryonic lens. Among these are 
the nuclear proto-oncogenes, c-myc, c-fos, and p53, and the membrane- 
associated tyrosine-specific protein kinase, c-src. A transient increase in 
the expression of the c-myc gene which occurs as the differentiating cells 
withdraw from the cell cycle suggests that this proto -oncogene may regulate 
some aspect of differentiation. The increased expression of c-myc has been 
shown to be primarily post -transcriptional, although a small increase in 
transcription has also been observed. The increase in c-myc expression which 
occurs during differentiation can be mimicked pharmacologically by agents 
which block the lipoxygenase pathway of arachidonic acid metabolism. 
Increased levels of c-myc mRNA, whether in differentiating cells or in cells 
treated with lipoxygenase inhibitors, are correlated with accumulation of mRNA 
for the heat shock protein, HSP70. 



PHS 6040 (Rev. 1/84) 



SPO ai4-SIB 



Ts: 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



IZOI EY 00126-07 LMDB 



PERIOD COVERED 



Orfohpr 1 . 1Q87 m cippf-pmbpr -^0. 1 q«« 



TITLE OF PROJECT (80 charactars or less. 77f/e must tit on one line Oerweon the boraers.) 

Crystallin Genes: Structure, Organization, Expressioii and Evolution 



PRINCIPAL INVESTIGATOR (List ottier professional personnel below the Principal investigator.) (Name, title, laboratory, and instituta attiilation) 

PI: Joram Piatigorsky Ph.D. Chief LMDB, NEI 



Others: Ana B. Chepelinsky Ph.D. 

David M. Donovan Ph.D. 

Robert A. Dubin Ph.D. 

John F. Klement Ph.D. 

Thomas Leitman B.A. 



Research Biologist 
IRTA Fellow 
Staff Fellow 
Staff Fellow 
HH Medical Student 



LMDB, NEI 

LMDB, NEI 

LMDB, NEI 

LMDB, NEI 

LMDB, NEI 



COOPERATING UNITS (if any) 



See next page 



LAB/BRANCH 

Laboratory of Molecular and Developmental Biology 



SECTION 



Section on Molecular Genetics 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 
12 



PROFESSIONAL: 

12 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects G (b) Human tissues D (c) Neither 

n (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Usa standarO unraducad type. Do not axcaed tha space prmtdad.) 

We have continued to study the crystallin genes and their expression in the 
cellular eye lens. Experiments have provided evidence that the chicken and 
mouse oA-crystallin gene use different cis-acting sequences and trans-acting 
factors to regulate their expression. Partial purification of embryonic 
chicken lens nuclear proteins that bind to the mouse aA-crystallin promoter 
has been achieved. More importantly, it appears as if it will be possible to 
isolate these putative regulatory proteins and their cDNAs . In contrast to 
the aA-crystallin gene, which is highly lens -specific, the mouse aB-crystallin 
genes were shown to be expressed in numerous non-lens tissues (heart, kidney, 
skeletal muscle), although to a lesser extent than in lens. Experiments using 
cultured cells and transgenic mice indicated that regulation of the qB gene 
resides in its 5' flanking sequence. The chicken ^Bl-crystallin promoter has 
been shown to be lens-specific in cultured cells; deletion mutants suggested 
the presence of a negative regulatory sequence (-436/-296) in this gene which 
may contribute to its tissue-specific expression. The concept of gene sharing 
was developed, which refers to the same gene encoding. both a lens crystallin 
and a metabolic enzyme. Our data indicate that argininosuccinate lyase is 
encoded by the 5 -crystallin genes (possibly only the S2 gene in the chicken) 
and a-enolase by the single r -crystallin gene in ducks and chickens. The r- 
crystallin/a-enolase gene in ducks is expressed about 25 times more strongly 
in the lens than in the liver. Finally, sequences of tryptic peptides were 
obtained and an anti- serum was raised to a synthetic peptide specific for a 
major crystallin (Jl) of the jellyfish eye lens. This initiates molecular 
studies on the crystallin of these ancient cellular lenses. 



PHS 6040 (Rev. 1/84) 



SPO (W**!! 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



ZOl EY 00251-01 LMDB i 

I 



TITLE OF PROJECT (30 characters or less. Title must tit on one line between tne txinMrs.) 

Regulatory elements of the aA-crystallin gene promoter 



PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Pnncipal Investigator) (Name. atle. laboratory, ana institute attiliation) 

PI: Ana B. Chepelinsky Ph.D. Research Biologist LMDB, NEI 



Others: Teresa Limjoco M.D. 

Eric Wawrousek Ph.D> 

Joram Piatigorsky Ph.D. 

Bernd Sommer Ph.D. 



Visiting Fellow 

Staff Fellow 

Chief 

Guest Worker 



LMDB, NEI 

LMDB, NEI 

LMDB, NEI 

LMDB, NEI 



COOPERATING UNITS (it any} 

Clive Dickson 



LAB/BRANCH 

Laboratory of Molecular and Developmental Biology 



Ph.D. Imperial Cancer Research Fund 
London, England 



SECTION 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

3 



PROFESSIONAL: 
3 



OTHER: 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects D (b) Human tissues i] (c) Neither 

n (a1) Minors 
D (a2) Interviews 



SUMMARY OF WORK (Usa standard unraducad type. Do not axcaad the space providad.) 

We have continued to characterize the cis -regulatory elements of the murine 
oA-crystallin promoter responsible for the lens-specific expression of this 
gene. Hybrid genes containing murine qA 5' flanking sequences and the gene 
coding for the bacterial enzyme chloramphenicol acetyltransf erase (CAT) were 
constructed and their expression studied in explanted chicken lens epithelia 
and in transgenic mice. Our results indicated the presence of a proximal 
(-88/+46) and a distal (-111/- 88) domain which must interact for promoter 
function in the explanted chicken lens epithelia. The sequence -88/- 60 is 
essential for promoter function. The distal domain activates the proximal 
domain when placed at the 5' end but not when inserted at the 3' end of the 
CAT gene. The distal domain does not activate the enhancerless SV40 promoter. 
Point mutations indicated that bases at positions -108 and -109 are essential 
for the activating properties of the distal domain in explanted chicken lens 
epithelia. Experiments with transgenic mice showed that the sequence -88/+46 
directs CAT gene expression specifically to the lens. Gel retardation and 
methylation interference experiments provided evidence for selective binding 
of different embryonic chicken lens nuclear proteins to sequences -111/- 84 and 
-83/-55. The protein factor binding -111/-84 may have some similarities to 
the transcription factor Spl . 



PHS 6040 (Rev. 1/84) 



GPO 9I4-»I« 



y 



ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October 1, 1987 - September 30, 1988 

REPORT OF THE CHIEF, LABORATORY OF RETINAL CELL AND MOLECULAR BIOLOGY 

Gerald J. Chader, Ph.D. 



The mission of the Laboratory of Retinal Cell and Molecular Biology 
is to investigate the functioning of the neural retina, at the levels of 
both cell and gene functioning. To best achieve this goal, investigators 
in the Laboratory are grouped in three Sections, although there is a 
great deal of communication and collaboration between the groups. 

Following are some of the accomplishments of the Laboratory members 
in this past year: 

Section on Cell Biology: A possible defect in phospholipid 
metabolism has been uncovered in a canine model of inherited retinal 
degeneration. Palmitic acid incorporation is abnormal in affected dogs, 
suggesting a significant reduction in the ester if ication of palmitic 
acid in this disease. Another important finding is that the alkylating 
agent NMNN can induce a progressive retinal degeneration in test animals. 
This effect appears at the gene level. Thus, two important leads have 
been uncovered in approaching genetic and toxicologically-induced 
degenerative conditions of the neural retina. 

Section on Biochemistry: Members of this Section have also studied 
animal models of retinal degeneration. In this case, the hereditary 
models used were in mouse, cat and dog. Interestingly, an early defect 
in the secretion of the photoreceptor protein IRBP, interphotoreceptor- 
binding protein, was found. The rd gene in particular appears to ,code 
for this secretion defect. A related project of investigators in this 
Section is the study of animal models of human uveitis. With collabo- 
rators in the Laboratory of Immunology, the IRBP protein has been found 
to be highly uveitogenic, inducing a severe inflammatory condition in the 
eyes of mouse, rat and monkey. Moreover, small peptide fragments of the 
IRBP molecule have been pinpointed that cause the disease. This is a 
potentially major breakthrough that may allow for modes of therapy to be 
developed in the future. 

Section on Gene Regulation: This group has been very successful in 
investigating the IRBP gene. The entire bovine genomic IRBP has been 
cloned and fully sequenced. The protein is large, the mRNA is also large 
but the gene is relatively compact. The full amino acid sequence has 
been deduced from the nucleotide sequence; it has given clues as to many 
of the interesting functional domains in the IRBP molecule. For example, 
it can be seen that the protein is composed of four similar units, two of 
which may cooperate to bind a retinoid molecule. The four-fold repeat 
strongly indicates gene replication during evolution. These findings 
will make it possible to begin the study of gene expression of IRBP in 
test systems in the near future. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES ■ PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT MjMBER 

ZOl EY 00070-11 LRCMB 



PERIOD COVERED 

October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (BO characters or less. Title must tit on one line tmtween the txirders.l 

Vitamin A and Ocular Tissues 



PRINCIPAL INVESTIGATOR (Ust other prolessional personnel Oelow the Principal Investigator) (Name, title, laboratory, ana institute attillatlon) 



PI: Barbara Wiggert Ph.D. 

Others: Ling Lee M.S. 

Michael Redmond Ph.D. 

Gerald J. Chader Ph.D. 



Head, Section on 

Biochemistry 
Chemist 
Staff Fellow 
Chief 



LRCMB, NEI 

LRCMB, NEI 

LRCMB, NEI 

LRCMB, NEI 



COOPERATING UNITS (it any) 

LSU Eye Center, New Orleans, LA (N. Bazan, B. Scott); Johns Hopkins University, 
Baltimore, MD (R. Adler); University of Lund, Lund, Sweden (T. van Veen); 
University of Illinois College of Medicine, Chicago, IL (D. Pepperberg, H. Ripps) 



LAB/BRANCH 

Laboratory of Retinal Cell and Molecular Biology 



SECTION 

Section on Biochemistry 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 



2.7 



PROFESSIONAL: 



1.7 



OTHER: 



1.0 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (a1) Minors 
n (a2) Interviews 



El (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Usa standard unreduced type. Do not exceed the space provided.) 

Interphotoreceptor retinoid binding protein (IRBP) was studied in retinae of 
mice with allelic combinations at the rd and rds loci. Until postnatal day 7 
(P7), IRBP is located intracellular ly in all retinae. Thereafter, in the normal 
retina, IRBP increases and is found primarily in the interphotoreceptor matrix. 
In the rd/rd, +/+, and rd/rd, rds/rds mutants, IRBP drops rapidly after Pll and 
is not secreted but is present intracellularly during the remaining degenerative 
process. In the rd/rd, rds/rds mutant, IRBP loss significantly precedes visual 
cell loss. In contrast, retinae of rodless +/+, rds/rds and +/+, rds/+ mutants 
synthesize essentially normal amounts of IRBP until very late in the degenera- 
tive process when there is then a significant amount of intracellular IRBP. We 
conclude that abnormality in secretion combined with other factors could lead to 
the degenerated phenotype in mice bearing the rd gene. 

Four synthetic peptides based on amino acid sequences present in cyanogen 
bromide peptides of IRBP were shown to induce autoimmune uveitis (EAU) and 
pinealitis (EAP) in Lewis rats. One of these peptides, containing 23 amino 
acids, was highly immunopathogenic and also immunodominant. The other peptides 
were substantially less immunopathogenic and also non-dominant. 

IRBP was found to provide efficient delivery of retinol to the pigment 
j epithelium for esterif ication and storage in the eyecup of dark adapted toads 
j (B. Marinus). Purified bovine IRBP was found to be capable of binding exogenous 

radiolabeled docosahexaenoic acid and palmitic acid. 



PHS 6040 (Rev. 1/84) 



GPO 9t4>ail 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PWOJEC"' NUMBER 



!Z01 EY 00015-23 LRCMB 



I PERIOD COVERED 

i October 1, 1987 to September 30, 1988 



TITLE OP PROJECT IBO cnamermrs or •«. Tme mutt m on on» an* o«fw«*n in* oorotn.) 

The Cell Biology of the Vertebrate Retina 



PRINCIPAL INVESTIGATOR lUsi omtr onftannai ovmnnti omw me ^maoai mrmaigatot > mtme rnie. mooniory. ana maun attmatnni 

PI: Paul J. O'Brien 



Others : 



Sylvia B. Smith 
Caren C. Demars 



Ph.D. 


Head, Section on 

Cell Biology 


LRCMB, 


NEI 


Ph.D. 


IRTA Fellow 


LRCMB, 


NEI 


B.A. 


Biologist 


LRCMB, 


NEI 



COOPERATING UNITS « any) 



I lAB/BRANCm 

! Laboratory of Retinal Cell and Molecular Biology 



SECTION 

Section on Cell Biology 



IN5TTTUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-rEASS; 



PROFESSIONAL: 



i OTHER-. 



1.6 



0.9 



0.7 



J 



CHECK A«wRO»RiATE BOX/ESi 

^ (a) Human suDiects 
[I (al) Minors 
Z ia2) Imerviews 



LJ (b) Human tissues 



Lx (c) Neither 



SUMMARV OF WORK IJm i 



: jfimeuemo lynt. Oo net i 



The post-translational modifications of rhodopsin include acylation, 
giycosyiation and chromophore addition. All appear to take place in the rod inner 
segment. The resulting molecules exhibit a slightly higher molecular weight than 
the mature rhodopsin in the outer segment and thus can be distinguished. 
The role of the palinitate residues is unknown but could be related to membrane 
assembly. The addition of the vitamin A chromophore seems to be essential for 
intracellular transport of the opsin protein to the Golgi and to the outer 
segments. The addition of several sugar residues in the Golgi complex may be a 
requirement for noirmal outer segment disc formation since the rhodopsin molecules 
in the plasma membrane and basal folds have a higher molecular weight than 
rhodopsin in disc membranes. 

Rod outer segments contain a molecule with both inositol and glucosamine. 
This molecule is reminiscent of the phosphatidylinositol-glycan anchor found in 
transiently membrane bound proteins and may indicate the existence of a 
phospholipase mediated release mechanism. 

A manganese-dependent 5 ' -nucleotidase that cleaves cytidine monophosphate has 
been found to become highly active in rod outer segment tips at the time of disc 
shedding. It has been isolated, partially purified and characterized and could 
provide insight into new mechanisms related to the shedding process. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 

NOTICE OF INTRAMURAL RESEARCH PROJECT 



=bcj£: 



-.uMBE = 



1 




ZOl EY 00016-21 LRCMB' 


1 PERIOD COVERED 

i October 1, 1987 to September 30, 


1988 


1 



TITLE OP PROJECT 180 cnmmcmn v wu Tim must « on on* m oarxwvn rf>» o u m& n .i 

The Biochemistry of Normal and Dystrophic Retinas 



PRINCIPAL INVESTIGATOR lusi om»r oronuionai ovtonnw omtow m* (>nnaD»i imimtugmtor i iNmmt. om. imoormiory, mno msmun amimooni 

PI: Paul J. O'Brien 

Others: Sylvia B. Smith 

Caren C. Demars 



Ph.D. 


Head, Se'ction on 

Cell Biology 


LRQIB , 


NEI 


Ph.D. 


IRTA Fellow 


LRCMB, 


NET 


B.A. 


Biologist 


LRCMB, 


NEI 



COOPERATING UNITS (I myi 

School of Veterinary Medicine, University of Pennsylvania (G. Aguirre) 



LAB/BRANCH 

Laboratory of Retinal Cell and Molecular Biology 



SECTION 
Section on Cell Biology 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-VEARS: 



1.1 



PROFESSIONAL: 



0.9 



OTHER: 



0.3 



CHECK APPROPRIATE BOXTES) 

G (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



C (b) Human tissues Q (c) Neither 



SUMMARY OP WORK (Urn I 



I umaeucM lypa. Oo not aaeaae m* i 



This project examines biochemical events unique to the retina, particularly 
the synthesis and modification of photoreceptor membrane components, in the retinas 
of vertebrates which can be affected by inherited retinal degenerations. The 
synthesis of the visual pigment, rhodopsin, occurs at a normal rate as measured by 
radioactive leucine incorporation following intravitreal injection in the eyes of 
miniature poodles affected with progressive rod-cone degeneration. Similarly, 
the glycosylation and acylation of rhodopsin were found to be normal following 
intravitreal injection of labeled fucose or palmitic acid, respectively. 
However, phospholipid synthesis or degradation, measured by radioactive palmitic 
acid incorporation, appears to be different in the affected dogs, suggesting a 
possible metabolic defect in this inherited disorder. The evidence suggests a 
significant diminution in the ester if ication of palmitic acid. Incubation of 
trephine punches of retina with labeled precursors produces the same labeling 
pattern in phospholipids as does intravitreal injection. Thus many precursors can 
be screened with a single retina. 

Transplacental exposure to the DNA alkylating reagent N-methyl-N-nitrosourea 
on day 16 of gestation in CD-I albino mice induces a progressive retinal 
degeneration beginning at 4-6 weeks of age. No obvious defect in either protein or 
phospholipid synthesis can be demonstrated. Thus a more subtle defect may have 
! occurred such as the alteration of a small number of genes . 



Due HAAA fC«v •1U1 



r,mn *■* «.•»■ 



1 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT iMUMBER 



ZOl EY 00148-15 LRCMH 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must lit on one line Between the txiraers.) 

Visual Control Mechanisms 



PRINCIPAL INVESTIGATOR (Ust other prvtessionai personnel below the Pnnapal Investigator.) (Name, title, laboratory, ana institute attillaeonl 
PI: Gerald J. Chader Ph.D. Chief LRCMB, NEI 

Others: R. Theodore Fletcher M.S. Chemist LRCMB, NEI 



COOPERATING UNITS (it any) 

School of Veterinary Medicine, University of Pennsylvania (G. Aguirre); Department 
of Anatomy, Erasmus University, Rotterdam, The Netherlands (S. Sanyal); Department 
of Zoology, University of Lund, Lund Sweden (T. van Veen) 



LAB/BRANCH 

Laboratory of Retinal Cell and Molecular Biology 



SECTION 

Section on Gene Regulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 



0.7 



PROFESSIONAL: 



0.2 



OTHER: 



0.5 



CHECK APPROPRIATE BOX(ES) 

G (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



[3 (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Us» siandartl urtrvOuctd typo. Do not •xcaatf tfw spaca provided.) 

Several diseases appear to only strike the neural retina. Thus, there may be 
important proteins or other substances that are specific to the retina and which 
are abnormal either in function or concentration in these retinal diseases . Such 
a protein may be IRBP, the interphotoreceptor retinoid-binding protein. We have 
found a greatly decreased concentration of this protein in an early stage of 
hereditary retinal degeneration in the Abyssinian cat. Other proteins may not be 
retina-specific but possible defects in their synthesis and/or function may 
particularly affect retinal metabolism. Such a protein, the cAMP-dependent 
protein kinase, is found in many cell types but appears to have a defect in 
synthesis in retinoblastoma t\unor cells grown in tissue culture. Such a defect 
could cause or contribute to the uncontrolled growth of retinoblastoma cells in 
vitro and perhaps in vivo as well. 



PHS 6040 (Rev. 1/84) 



CPO 9l4>ail 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00124-08 LRCMB 



PERIOD COVERED 
October 1, 1987 to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must lit on one line Oefwoen the borders.) 

Metabolism of the Retina and Pigment Epithelium 



PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Phnapal Investigator) (Name, title, laboratory, and institute affiliation) 

PI: Gerald J. Chader Ph.D. Chief LRCMB, NEI 



Others: Robert Waldbillig Ph.D. 
R. Theodore Fletcher M.S. 
Dagmar Arnold M.D. 



Expert 

Chemist 

Visiting Associate 



LRCMB, NEI 
LRCMB, NEI 
LRCMB, NEI 



COOPERATING UNITS (it any) 



LAB/BRANCH 

Laboratory of Retinal Cell and Molecular Biology 



SECTION 

Section on Gene Regulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 



2.3 



PROFESSIONAL: 



1.8 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



OTHER: 



0.5 



El (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Usa atanOam unreOucad type. Oo not axeted tfw space provided.) 

Low molecular weight, soluble factors as well as extracellular matrix molecules 
play major roles in the growth and development of all tissues. This includes 
normal tissue and tumor tissue. Laminin may play such a critical role in retinal 
development. Insulin and especially IGF-1 may act as messengers coding for 
differentiation in the retina and, by affecting phosphorylation of the G-protein 
transducin, may be directly or indirectly involved in the visual process. 
Abnormal protein kinase activity and thus cyclic AMP function may be involved in 
the rapid, uncontrolled growth of retinoblastoma tumor cells. 



PHS 6040 (Rev. 1/84) 



SPO 914-CII 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 

PERIOD COVERED 

October 1, 1987 to September 30, 1988 
TITLE OF PROJECT (BO characters or less. Title must lit on one line between tne Borders. I 

Molecular Genetics of the Eye and Ocular Diseases 



PROJECT NUMBER 

ZOl EY 00196-05 LRCMB 



PRINCIPAL INVESTIGATOR (List other pmtassionat personnel Below tne Pnncipal Investigator) (Name, title, laboratory, ana institute atftliation) 
PI: John M. Nickerson Ph.D. Biologist LRCMB, NEI 



Others: Diane Borst Ph.D. 

T. Michael Redmond Ph.D. 

Jing-Sheng Si M.D. 

Adriana Albini Ph.D. 

Lila Inouye M.D. 

Judith Toffenetti Ph.D. 



IRTA Fellow 
Staff Fellow 
Visiting Associate 
Visiting Associate 
Staff Fellow 
Staff Fellow 



LRCMB, NEI 

LRCMB, NEI 

LRCMB, NEI 

LRCMB, NEI 

LRCMB, NEI 

LRCMB, NEI 



COOPERATING UNITS (if any) 



See next page. 



LAB/BRANCH 

Laboratory of Retinal Cell and Molecular Biology 



SECTION 

Section on Gene Regulation 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 



6.2 



PROFESSIONAL: 



6.2 



OTHER: 



0.0 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
n (a1) Minors 
D (a2) Interviews 



(il (b) Human tissues G (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 

My laboratory has isolated and characterized recombinant DNA molecules necessary 
for the study of the structure and expression of IRBP ( Interphotoreceptor 
Retinoid-Binding Protein). We have cloned many different cDNAs (copies of the 
IRBP messenger RNA) from bovine and human retina. We have screened a human 
retina cDNA library with the bovine IRBP cDNA probe and have identified several 
large cDNA clones up to 3.5 kb in length for human IRBP. We have sequenced 
portions of all of these overlapping cDNA clones. The IRBP mRNA is long, 4.4 to 
7.4 kb in several species and usually gives only one band on a Northern blot. 
The cDNA and gene sequences have been used to predict the amino acid sequence of 
the protein. The polypeptide contains four 300 amino acid long repeats, with 30- 
40% identity among the repeats. These sequences have been helpful in the 
analysis of the uveitogenic peptides in IRBP. DNA sequence analysis of the gene 
clone has identified the authentic N- terminus, the putative initiator methionine 
codon, a putative pro-peptide and a putative signal peptide sequence of the IRBP 
polypeptide. The chromosomal location of the IRBP gene is: 10 for human, 4 for 
dog, and 11 for mouse. The bovine gene structure is compact for the size of the 
protein, and has only 3 introns. The stmicture of the gene suggests an 
interesting evolution, involving a processed gene intermediate and two unequal 
crossovers. 



PHS 6040 (Rev. 1/84) 



GPO ■I4>»lt 



ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October 1, 1987 - September 30, 1988 

ElEPORT OF THE CHIEF, LABORATORY OF SENSORIMOTOR RESEARCH 
Robert H. Wurtz, Ph.D. 



This is the Tenth Annual Report of the Laboratory of Sensorimotor Research. 
Rather than summarize last year' s work (which is detailed in the following indi- 
vidual annual reports) I would like to outline the progress of the Laboratory in 
its first decade. Even this unusually long report describes only major themes, 
omitting a series of other important areas of work within the laboratory. 

The investigators in this Laboratory share an interest in the brain mechan- 
isms underlying vision and eye movement . Three fields of neurophysiology that 
are particularly well developed relate to the control of eye movements: the pro- 
cessing of visual target information, the generation of eye muscle innervation, 
and the adaptive maintenance of adequate performance. Knowledge in these fields 
has advanced rapidly over the last twenty years, and members of the Laboratory 
have been at the forefront of each field. Despite this progress, one of the 
great unresolved problems in neurophysiology remains: how does sensory informa- 
tion give rise to motor responses? One of the goals of the Laboratory has been 
to study not only the individual aspects of visual and motor processing by the 
brain, but also the transition from visual to motor signals. 

The visual and oculomotor functions of the brain that we study have been 
shown to be similar in hiunans and old world monkeys ( Macaca mulatta) so that our 
experiments on the monkey serve as a model for humans. Behavioral, physiologi- 
cal, and anatomical experiments that are possible in the monkey have given us our 
most fundamental understanding of how visual and oculomotor functions are likely 
to be organized in humans. In addition, several investigations in the laboratory 
illustrate how the precise analysis possible in the visual-oculomotor system has 
allowed exploration of more general questions of brain research. 

One of the major advantages of studying this visual-oculomotor system is 
that this system consists of a series of single movement stibsystems, all 
integrated to produce a coordinated system, but each sufficiently separated to 
allow each to be studied individually. Work in the laboratory has concentrated 
on a number of these movement systems including the saccadic, pursuit, and ocular 
following systems. 

Saccadic eye movements . These movements shift the direction of the eye 
rapidly from one part of the visual field to another to bring the fine-grained 
fovea of the retina onto the area of the visual field of interest . This is the 
system whose integrity is critical for reading and for the frequent inspection of 
our surroundings . 

Dr. Michael Goldberg has concentrated on an understanding of the saccadic 
system at the highest level of organization, the frontal region of the cerebral 
cortex. In an area that is referred to as the frontal eye fields, he has 



identified a set of neurons that are active during different phases of the sac- 
cadic eye movement including cells responding to visual stimuli, cells discharg- 
ing in association with purposive eye movements, and cells discharging after the 
occurrence of an eye movement . He has found that the cells discharging in rela- 
tionship to eye movements represent the major output of this cortical area to the 
brainstem structure related to saccadic eye movements, the superior colliculus. 
Removal of these cortical cells by selective lesion has revealed that a signifi- 
cant function of the area is the generation of saccadic eye movements under com- 
plex conditions, e.g., saccades made to the location of a remembered target. 

Work on the frontal eye fields has dealt with one of the most fundamental 
problems that the brain must solve in controlling movement: the conversion of a 
sensory error into an accurate motor movement. For saccadic eye movements, this 
question is one of how the brain converts the difference between where the eye is 
looking and where the desired target is located into the spatial coordinates used 
to guide the eye movement . Most solutions to this problem hypothesize a spatial 
map within the brain, but only rudimentary spatial maps have been found. On the 
basis of his experiments. Dr. Goldberg developed an alternate hypothesis that 
argued that the brain uses only the difference in eye and target position but 
updates this difference information after every eye movement . All the elements 
necessary for this system have been identified in the activity of single cells in 
the frontal cortex. Thus, the work on the frontal eye fields has produced impor- 
tant hypotheses about the way the brain solves fundamental sensory motor problems 
and represents the most quantitative and detailed study of one of the highest 
levels of cortical function. Insights gained from this work have recently led to 
a method of treatment of patients whose reading is interrupted by extraneous sac- 
cades . 

An area of the basal ganglia in the brainstem (the substantia nigra pars 
reticulata) receives projections from frontal cortex, and Dr. Hikosaka and I 
discovered that cells in this area that decrease their discharge in relation to 
saccades to visual targets or with saccades to locations that had to be remem- 
bered. Since the output of this structure has been demonstrated to be inhibitory 
on the next stage of the saccadic system, the superior colliculus, it is likely 
to exert a control on the superior colliculus not previously realized. We subse- 
quently demonstrated such control by blocking or mimicking the action of the 
inhibitory transmitter, GABA in the pathway to the superior colliculus. Because 
of the precision of recording of saccadic eye movements and the control of the 
conditions under which they are made, this oculomotor-related pathway is probably 
the best understood output of the basal ganglia. Sxibsequent tests in humans with 
a disease of the basal ganglia (Parkinsons Disease) revealed some of the same 
deficits seen in the monkey during the treatment with drugs that mimic GABA. 

A target of both the frontal eye fields and the substantia nigra is the 
superior colliculus and its relation to saccadic eye movements was first 
described by Goldberg and me nearly 20 years ago. Subsequent work in our labora- 
tory and many others has contributed to defining the role of cells in the supe- 
rior colliculus to the control of saccadic eye movements and the consequences of 
damage of the structure. The classic understanding of the colliculus has been 
that it has provided information on the motor error, the difference between posi- 
tion of the eye and the target. Work in the laboratory in the last several years 
has shown, however, that there are additional cells in the superior colliculus 
that provide information about how far the eye has gone toward reaching that tar- 
get, a dynamic motor error. 



One test of the completeness of the knowledge in a field is the ability to 
make mathematical models that perform realistically. Using knowledge common to 
the field, and the results of recent experiments within the Laboratory, Dr. Lance 
Optican has been able to develop a new model that incorporates both visual and 
motor elements of the saccadic system. This model incorporates physiological 
observations and produces dynamically realistic eye movements when simulated on a 
computer. Dr. Optican' s model is a unique achievement in its successful descrip- 
tion of how visual information may control saccades . The advantage of this 
modeling approach is that it suggests a new concept of the organization of the 
brain stem control of saccades, incorporates both new and old physiological 
observations, and reconciles seeming discrepancies among different experimental 
results. The model emphasizes the importance of some of our new observations on 
the superior colliculus, and has redirected study of the role of the superior 
colliculus in controlling eye movements throughout the field. 

Our knowledge of the saccadic system is sufficiently extensive that it has 
warranted a volume in Reviews of Oculomotor Research, edited by me and Dr. Gold- 
berg. 

Pursuit eye movements . These movements allow the fovea to be directed at a 
target moving in the visual field, and among mammals this system is most highly 
developed in primates, including humans. An understanding of this system is 
dependent on an understanding of visual motion processing within the brain, which 
in primates is largely concentrated within the cerebral cortex. Work by me and 
my collaborators has capitalized on the identification of different cortical 
areas in front of the primary visual area, particularly areas MT and MST, where a 
high proportion of cells are sensitive to visual motion. We found that punctate 
chemical lesions of MT led to a deficit in pursuit but not saccades; this 
represents the clearest demonstration to date that an area of visual cortex can 
be related to one type of visual processing (motion) but not for another (posi- 
tion) . Cells in MST provide both visual motion information and added non-visual 
information on direction of pursuit eye movements. Discrete damage to this area 
produces a deficit in pursuit toward the side of the brain with the lesion, as 
has been classically observed following damage to parietal cortex in humans. 

Ocular Following Movements . Several types of eye movements have long been 
recognized to reduce slippage of the retinal image in order to provide clear and 
stable vision in spite of movements of the head and body: the vestibular-ocular 
response and the optokinetic response. Dr. Frederick Miles has now identified an 
entirely new visual-motor response not previously recognized that also aids in 
maintaining clear vision, and he has referred to this as an ocular following 
response. He has found in the monkey that this response has an incredibly short 
and regular latency close to 50 msec, and that it is generated by motion of the 
visual field. The sensitivity of the system is increased shortly after a sac- 
cade, and could serve to minimize in our normal con^lex environment the drifts of 
eye movement that follow saccades. Stibsequent experiments have revealed a simi- 
lar though not as robust ocular following response in humans . Through a series 
of ingenious experiments on the monkey. Dr. Miles and his collaborators have been 
able to dissect out the variables affecting this response and have suggested that 
the ocular following response is designed for stabilization of the visual scene 
during translation through the environment . This is in contrast to the optok- 
inetic and vestibulo-ocular systems which steibilize the visual scene during rota- 
tion of the head and body. The recognition of this control system raises the 
possibility that a number of characteristics ascribed to other ocular motor 



systems, such as the pursuit system, are actually part of this newly recognized 
translational control system. Dr. Miles' experiments also illustrate further the 
power of a carefully detailed behavioral analysis applied to a complex system 
within the brain. 

Visual Selection . The selection of a target from among the myriad of those 
available is important for several forms of behavior and critical for the execu- 
tion of saccadic eye movements. Dr. David Lee Robinson has concentrated on the 
neural basis of this selection process and more generally on visual attention 
independent of the direction of gaze. His work has concentrated on the pulvinar 
nucleus, a visual area in the thalamus, and much of our knowledge of the function 
of this structure is the result of his investigations. In studies of the pulvi- 
nar, superior colliculus, and parietal cortex, he has shown the modified 
responses of single cells while the monkey shifted attention from one part of the 
visual field to another, and has also been able to use small reversible chemical 
lesions to reveal the contribution of these structures to shifts of attention. 
These experiments not only demonstrate the ability to relate a brain structure to 
such a high level function as selective attention, but also show for the first 
time a function for the pulvinar, a hitherto puzzling thalamic structure. These 
insights into attentive processes have led to investigations of patients with 
diseases affecting the parietal and frontal regions of the cerebral cortex and 
progressive supranuclear palsy; such experiments reveal that different types of 
deficits are associated with damage to different regions of the brain. 

Adaptive Control . The oculomotor systems, in order to function properly, 
must be continually adjusted for changes that occur normally in the course of 
developing and aging or that result from diseases affecting the system. Adjust- 
ment of these oculomotor systems therefore require adaptive control to maintain 
their precision, particularly if the system usually operates "open loop", that 
is, information about any error in the movement arrives in the brain too late to 
alter that movement. Dr. Miles and Dr. Optican have been leaders in investigat- 
ing adaptive control in the oculomotor system. Before joining the laboratory. 
Dr. Miles had studied extensively the cellular changes related to the adaptive 
changes of the vestibular ocular reflex as well as the conditions under which 
this adaptation occurred, and since joining the laboratory he has shown that the 
plasticity of this system is so specific that adaptation can occur for certain 
frequencies of vestibular stimulation but not others. He and Dr. Optican showed 
that the amplitude of saccades and the stibsequent ocular drifts were also subject 
to adaptive control. Subsequent work by Dr. Miles has revealed for the first 
time the adaptive control of vergence accommodation, and of the newly identified 
ocular following response. Dr. Optican was the first to demonstrate the adaptive 
control of the pursuit system. This was an important finding, since the pursuit 
system is not "open loop", i.e., pursuit movements are slow enough that they 
influence their input (retinal slip) and can provide adequate control. In this 
case, the adaptive control was designed to proved not merely adequate, but 
optimal, performance, and this finding raises the possibility that all neural 
systems are under adaptive control. The role of the cerebellum has been demon- 
strated in several of these cases of adaptation, giving this structure a major 
role in the adaptive control of eye movements, and probably a more explicit func- 
tion than that postulated for any other system. 

Visual Coding . While work in the laboratory has centered on visual-motor 
control, a number of experiments have concentrated on visual processing, particu- 
larly in the visual pathways from primary visual cortex into extrastriate areas 



related to visual motion (MT and MST) and to areas presumably related more to the 
analysis of form (inferotemporal cortex) . Salient among these investigations has 
been the work, by Dr. Optican in collaboration with Dr. Richmond of the NIMH which 
questions the fundamental assumption of nearly all studies of the visual system 
that single neurons convey information only by the strength of their discharges . 
In their investigations on inferotemporal and striate cortex neurons, they have 
been able to show that the pattern of discharge is critical, and that the tem- 
poral modulation of the cell discharge contains roughly double the information 
transmitted by a neuron as compared to the total number of spikes alone. Drs. 
Optican and Richmond have developed hypotheses about how the visual system might 
encode within one neuron a series of visual characteristics of a stimulus. Their 
hypotheses raise fundamental questions about the way in which the brain codes 
visual information and additional fundcimental questions about the organization of 
the visual system based on the notions of the tuning characteristics of indivi- 
dual neurons that has grown out of the work of Hubel and Wiesel in the last 25 
years. Investigation of their hypothesis will yield fundamental insights on some 
of the most intriguing questions related to the visual system, namely, how such 
properties as form, color, and motion are represented by neurons within the 
brain. 

The future challenge . In comparison to the challenge of understanding these 
elegant and precise visual and visual-motor systems within the brain, our pro- 
gress has been modest . But in comparison to the knowledge that we had about 
these systems 10 years ago, I find our progress very gratifying. Because of the 
relative simplicity of the oculomotor system we probably now understand its sub- 
systems better than any other other system in the primate, and the field of ocu- 
lomotor control is the first within neurophysiology to be on the brink of under- 
standing the entire flow of information- from the visual sensation to the motor 
response. We look on the visual-motor function of the brain as providing clues 
to higher brain function. The number of fundamental problems already studied 
that relate to general issues of brain function indicate, I think, that this 
approach is successful. One of the most exciting challenges facing the Labora- 
tory in the future will be to use our expertise in the study of visual, motor and 
adaptive neural mechanisms to produce a new field of sensorimotor physiology, one 
able to study the brain's systems as an integrated whole. 

Our laboratory has benefited greatly from the interactions of a group of 
senior scientists working on different but related problems, and we have been 
able to share intellectual challenges and technical break throughs quickly and 
efficiently. It is obvious that there is extensive overlap in our interests that 
has led to substantial cross fertilization in both directions of experiments and 
experimental design. At a technical level we have benefited from technical 
advances now used beyond our laboratory: the implantation of the eye coil (Judge, 
Richmond and Chu) , the Rex laboratory computer software, (Hays, Richmond and 
Optican) , and the ASP model simulation software (Optican and Goldstein) . In the 
10 years since its organization, I think our laboratory has become preeminent in 
the study of the visual-motor system and, as a consequence, we are able to 
attract some of the most talented young investigators from throughout the world. 
I can only hope that the next 10 years will be as profitable as the last. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00049-10 LSR 



PERIOD COVERED 

October 1, 1987, to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must fit on one line between the borders.) 

Cerebral Cortical Mechanisms for Eye Movements and Visual Attention 



PRINCIPAL INVESTIGATOR (List other orotessionat personnel t>elow the Principal Investigator.) (Name, title, laboratory, ana institute atfiliation) 

PI: Michael E. Goldberg M.D. Chief, NMS LSR, NEI 



Others: Mark A, Segraves Ph.D. 

Edmond J. FitzGibbon M.D. 

Carol L. Colby Ph.D 

Jean-Rene Duhamel Ph.D. 



Senior Staff Fellow LSR, NEI 

Senior Staff Fellow LSR, NEI 

Guest Researcher LSR, NEI 

Visiting Scientist LSR, NEI 



COOPERATING UNITS (if any) 



LAB/BRANCH 

Laboratory of Sensorimotor Research 



SECTION 

Neuro-Ophthalmologic Mechanisms Section 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

3.6 



PROFESSIONAL: 

2.5 



OTHER: 



1.1 



CHECK APPROPRIATE BOX(ES) 

D (a) Human subjects 
D (a1) Minors 
D (a2) Interviews 



D (b) Human tissues K! (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 

The functional nature of the projection from the frontal eye field to the brain 
stem has been studied in the rhesus monkey. Like the frontotectal projection, the 
frontopontine projection contains cells which discharge in association with eye 
movements or visual fixation, but not cells which have exclusive peripheral 
visual responses . 

The nature of the visual stimuli evoking smooth pursuit were was studied using 
open-loop visual methods , Superimposition of open-loop position and velocity 
errors during pursuit maintenance resulted in the generation of eye velocities 
that indicated that stimulus position as well as stimulus velocity is an impor- 
tant stimulus for the maintenance of smooth pursuit. 

The time course and dynamics of uniocular saccadic adaptation were studied in 
monkeys who were made to adapt to a weakened eye. At first the weakened eye had 
a hysteresis in orbital position, and an orbital-position-dependent saccadic 
inaccuracy. Both the hysteresis and the orbital position dependent effects were 
compensated for in a point by point manner with experience. The results suggest 
that the oculomotor system has a complicated and sensitive corrective mechanisms 
for the non-linearity of orbital mechanics . Any physical derangement causes 
maladjustment of this condensation, which can be adapted in time. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00153-06 LSR 



PERIOD COVERED 

October 1, 1987, to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must tit on one line between the boraers.) 
Visual Motion and the Stabilization of Gaze 



PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, ana institute affiliation) 



PI: 



Frederick A. Miles 



Others : Hubert Kimmig 
Urs Schwarz 



D.Phil 

M.D. 
M.D. 



Chief, OCS 

Visiting Fellow 
Visiting Fellow 



LSR, NEI 

LSR, NEI 
LSR, NEI 



COOPERATING UNITS (if any) 



Joshua Wallman 



Ph.D. 



Professor 



CUNY 



LAB/BRANCH 

Laboratory of Sensorimotor Research 



SECTION 

Oculomotor Control Section 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS; 

1.8 



PROFESSIONAL: 

1.0 



OTHER: 



0.8 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects 
D (al) Minors 
n (a2) Interviews 



n (b) Human tissues K] (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 

Processes important for emmetropization, whereby the optical power of the eye 
comes to match its size, were examined in developing chicks. The eyes of chicks 
raised in a low-ceiling environment were significanctly more myopic in the upper 
field than the eyes of control animals. Most of this effect could be accounted 
for by selective local increases in the depth of the posterior chamber. This is 
consistent with the notion that vision plays an active role in sculpting the 
chick' s eye to achieve appropriately f ocussed retinal images in the different 
paijts of the visual field. The maintenance of stable retinal images was studied 
in chicks by examining the visual mechanisms responsible for stabilizing the 
head. The head movements induced by translation or rotation of the surroundings 
revealed powerful stabilizing reflexes that seem to be mediated by separate 
mechanisms, e.g., responses to translational disturbances showed none of the 
naso-ten^oral asymmetries characteristic of the ocular stabilization mechanisms 
in birds that deal with rotations of the surroundings. Further, rotational 
oscillations of the surroundings at high frequencies evoked lateral translations 
of the head rather than rotations, suggesting that only the translational mechan- 
isms respond over this part of the range. Image stabilization was also studied 
in monkeys by examining the visual mechanisms underlying their ocular pursuit of 
small moving targets. The early suppression of ocular pursuit by featured back- 
grounds, described by Keller & Khan (1986) , was shown not to be due simply to the 
reduced physical salience of the track target: suppression was still" seen, albeit 
i reduced, if the path of the target was devoid of features and consisted of a dark 
band. In fact, suppression was still evident even when the band was 30° wide. 
Suppression also showed interocular transfer, whereby texture seen only by one 
eye could suppress pursuit initiated by target motion seen only by the other eye. 
This indicates that suppression can result entirely from centrally mediated 
interactions between visual inputs . 



PMQ firiAr\ /Daw 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00152-06 LSR 



PERIOD COVERED 

October 1, 1987, to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title must tit on one line between the borders.) 

Adaptive Changes in Saccadic Innervation 



PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator ) (Name, title, latxjratory, ana institute affiliation) 



PI: 



Lance Opt i can 



Others : Zoi Kapoula 

Michael E . Goldberg 
David M. Waitzinan 
Terence P . Ma 



Ph.D. Res. Biomedical Engineer 

Ph.D. Guest Researcher 

M.D. Chief, NMS 

M.D. Staff Fellow 

Ph.D. Post-Doctoral Fellow 



LSR, NEI 

LSR, NEI 

LSR, NEI 

LSR, NEI 

LSR, NEI 



COOPERATING UNITS (if any) 



LAB/BRANCH 



Laboratory of Sensorimotor Research 



SECTION 

Oculomotor Control Section 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

2.3 



PROFESSIONAL: 



1.9 



OTHER: 



0.4 



CHECK APPROPRIATE BOX(ES) 

CS (a) Human subjects 
D (a1) Minors 
n (a2) Interviews 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 

Saccades are the rapid eye movements used to change visual fixation. These eye 
movements are very accurate and end without drift. One of the projects in this 
lab has studied the ability of the brain to control post-saccadic ocular drift in 
both eyes. We have found that human subjects, like monkeys, respond to 
optically-induced post-saccadic slip by developing post-saccadic ocular drift in 
a compensatory direction. This suggests that after normal saccades there should 
be no post-saccadic drift. However, normal subjects usually show post-saccadic 
ocular drift in one or both eyes after every saccade. Attempts to cause monocu- 
lar adaptation failed, suggesting that the drift after saccades in normal sub- 
jects can not be corrected because of the lack of an independent mechanism for 
each eye. 

Another study in this lab has focussed on the neural mechanisms of the sensory- 
to-motor transformation needed to turn visual target information into saccadic 
eye movements. It has long been known that the superior colliculus (SO) in the 
brain stem contains both visual and motor maps related to saccadic eye movements. 
Up until now it has been assumed by all that the colliculus was providing a 
static command signaling the change of eye position that would get the eye on 
target. This leaves unresolved the issue of how the command signal is 
transformed from a location, or cell-code in SC into the frequency/duration code 
needed by the eye muscles. Based on new experimental analyses of SC activity 
I patterns, we have formulated a radical new hypothesis of SC function. According 
to this hypothesis, the SC is the source of the dynamic motor error signal in a 
local feedback loop controlling saccades. By placing it in the loop, the SC is 
now shown to be part of the transformation from cell-coded to 

frequency/duration-coded signals. This radical new hypothesis has far reaching 
consequences for how we think about the neural control of saccadic eye movements. 



PHS 6040 (Rev. 1/841 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PERIOD COVERED 

October 1, 1987, to September 30, 1988 



PROJECT NUMBER 

ZOl EY 00045-10 LSR 



TITLE OF PROJECT (SO characters or less. Title must fit on one line Oefween rfte borders ) 

Visuomotor Properties of Neurons in the Thalamus 



PRINCIPAL INVESTIGATOR (List other professional oersonnel below the Principal Investigator.) (Name, title, laboratory, ana institute attiliationi 



PI: David Lee Robinson Ph.D. 

Others: Caroline Kertzman Ph.D. 

Richard Sherins M.D. 

Irene Litvan M.D. 

Edmond FitzGibbon M.D. 

James Carl M.D. 



Research Physiologist 

IRTA 

Res . Endocrinologist 

Clinical Fellow 

Sr. Staff Fellow 

Sr. Staff Fellow 



LSR NEI 
LSR NEI 
NICHD 
NINCDS 
LSR NEI 
LSR NEI 



COOPERATING UNITS (if any) 



LAB/BRANCH 

Laboratory of Sensorimotor Research 

SECTION 

Visuomotor Integration Section 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

2.3 



CHECK APPROPRIATE BOX(ES) 

[3 (a) Human subjects 
D (a1) Minors 
n (a2) Interviews 



PROFESSIONAL: 



1.5 



OTHER: 



0.8 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided ) ' 

We have studied the neural mechanisms of visual spatial attention in humans 
and monkeys. Both species fixated on a spot of light and responded with their 
hands to peripheral visual targets. Reaction times were faster for targets pre- 
ceded by a light (cue) on the same side (validly cued) than when the cuing light 
was on the opposite side (invalidly cued) . The performance of the monkeys was 
selectively altered by injection of transmitter-related drugs into the superior 
colliculus. Neurons recorded from the colliculus while the monkey performed this 
task responded well to both the cue and target when they were in the visual 
receptive field. For some cells when the cue was placed outside of the visual 
receptive field, the neuron still discharged (weakly) as if it were influenced by 
the movement of attention through its receptive field. 

We tested a population of males treated for Kallmann's syndrome. All 
responded faster than did age-matched control subjects. The controls were only 
able to equal the patients' performance when highly motivated. A subset of 
patients have synkinesis. This group performs the task as do subjects with 
lesions of parietal cortex. They are very slow responding to invalidly cued tar- 
gets to one side and responding to any targets after diffuse cues. Such data 
suggest that these patient have an undiagnosed dysfunction of the parietal cor- 
tex. 

When patients with progressive supranuclear palsy are tested, they are sub- 
stantially slower in all respects compared to control subjects. They have signi- 
ficantly increased differences between valid and invalid reaction times suggest- 
ing a slowing of the movement of attention. They are able to move their atten- 
tion vertically as well as horizontally, even though they cannot make vertical 
eye movements. Treatment with physostigmine, a cholinesterase inhibitor, reduces 
the difference in reaction times suggesting an improvement in the ability to move 
attention. The therapy had no effect on the oculomotor capacity of the patients 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 



ZOl EY 00109-08 LSR 



PERIOD COVERED 
October 1, 1987, to September 30, 1988 



TITLE OF PROJECT (80 characters or less- Title must tit on one line between tne boraers.l 
Visuomotor Processing in the Primate Brain 



PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator) (Name, title, laboratory, and institute affiliation) 



PI: Robert H. Wurtz 
Others: Hidehiko Komatsu 

Dwayne S. G. Yamasaki 
Jean-Pierre Roy- 
David M. Waitzman 
Terence P . Ma 
Lance M. Optican 



Ph.D. 


Chief 


LSR, 


NEI 


Ph.D. 


Visiting Scientist 


LSR, 


NEI 


Ph.D. 


Guest Researcher 


LSR, 


NEI 


M.D. Ph.D. 


Guest Researcher 


LSR, 


NEI 


M.D., Ph.D. 


Staff Fellow 


LSR, 


NEI 


Ph.D. 


Guest Researcher 


LSR, 


NEI 


Ph.D. 


Res. Biomed. Engineer 


LSR, 


NEI 



COOPERATING UNITS (if any) 



LAB/BHANCH 

Laboratory of Sensorimotor Research 



SECTION 

Visuomotor Integration Section 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 



20892 



TOTAL MAN-YEARS: 

4.3 



PROFESSIONAL: 



2.7 



OTHER: 



1.6 



CHECK APPROPRIATE BOX(ES) 

n (a) Human subjects 
n (a1) Minors 
n (a2) Interviews 



D (b) Human tissues S3 (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 

We have continued our study of the visuomotor processing in the brain for the 
generation of smooth pursuit and saccadic eye movements . In the study of smooth 
pursuit eye movements, we concentrated on an area of cerebral cortex devoted to 
the analysis of visual motion, MST. We found that by stimulating this area dur- 
ing pursuit, we produced an acceleration toward the side of the brain being 
stimulated. Added to our previous observations, these experiments have localized 
the brain region related to the maintenance of pursuit eye movements. We also 
determined that recovery of pursuit following damage to these areas was minimally 
affected by visual experience during the recovery period. In the study of sac- 
cadic eye movements, we identified a type of neuronal discharge in the superior 
colliculus that indicated that some cells in this structure receive information 
about how far the eye has moved during a saccade. This led to a reformulation of 
the role of the superior colliculus in the generation of saccadic eye movements, 
and a new model of saccadic control. 



DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBLIC HEALTH SERVICE 
NOTICE OF INTRAMURAL RESEARCH PROJECT 



PROJECT NUMBER 
ZOl EY 00244-01 LSR 



PERIOD COVERED 

October 1, 1987, to September 30, 1988 



TITLE OF PROJECT (80 characters or less. Title musf tit on one line between the boraers.) 

Oculomotor and visual disorders in himans 



PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, ana institute atiillationt 

PI: James R. Carl M.D. Senior Staff Fellow LSR, NEI 



Others: Edmond J. FitzGibbon M.D. 
Michael E. Goldberg M.D. 



Senior Staff Fellow LSR, NEI 
Chief, NMS LSR, NEI 



COOPERATING UNITS (it any) 



I LAB/BRANCH 

Laboratory of Sensorimotor Research 



SECTION 

Neuro-Ophthalmologic Mechanisms Section 



INSTITUTE AND LOCATION 

NEI, NIH, Bethesda, Maryland 20892 



TOTAL MAN-YEARS: 

0.5 



PROFESSIONAL; 



0.5 



OTHER: 



0.0 



CHECK APPROPRIATE BOX(ES) 

[S (a) Human subjects 
n (a1) Minors 
n (a2) Interviews 



n (b) Human tissues D (c) Neither 



SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) 

Eye movements were studied in patients with progressive supranuclear palsy. 
These patients were found to have abnormalities in fixation, pursuit and saccadic 
eye movements, with an uncoupling of the major trajectories of the horizontal and 
vertical components of oblique saccades. Vertical eye movements appeared possi- 
ble only when the horizontal system made small square wave jerks. 

Patients with a variety of forms of nystagmus were evaluated in an ongoing study 
of _the clinical significance of the nystagmus waveform. 

Eye movement recordings were also used to document early neurologic involvement 
in patients with xeroderma pigmentosum. 



ANNUAL REPORT 

NATIONAL EYE INSTITUTE 

October 1, 1987 - September 30, 1988 

REPORT OF THE ASSOCIATE DIRECTOR FOR BIOMETRY AND EPIDEMIOLOGY 

Daniel Seigel, Sc.D. 



Organization : 

The Biometry and Epidemiology Program consists of a Clinical Trials 
Branch, an Epidemiology Branch, and a Biometry Section. Drs. Frederick Ferris 
III and Robert Sperduto serve as Chiefs of the two Branches, respectively; Dr. 
Roy Milton is the Head of Biometry. Dr. Daniel Seigel is Associate Director. 



Functions : 

The Biometry and Epidemiology Program (BEP) has three main functions: 
research, education, and consultation. 

Research is the dominant function. , It is the Program's mission to plan, 
develop, and carry out human population studies concerned with the causation, 
prevention, and treatment of eye disease and vision disorders, with emphasis 
on the major causes of blindness. This includes studies of incidence and 
prevalence in defined populations, prospective and retrospective studies of 
risk factors, natural history studies, clinical trials, genetic studies, and 
studies to evaluate diagnostic procedures. 

Education : The BEP carries out a program of education in biometric and 
epidemiologic principles and methods for the vision research community. This 
program consists of courses, workshops, a fellowship program for 
ophthalmologists, publications, and consultation and collaboration on 
research. 

Consultation : The Program provides biometric and epidemiologic 
assistance to National Eye Institute intramural and extramural staff and to 
vision research workers elsewhere. The assistance ranges from consultation 
through collaboration as co-investigator. 

Research Activities ; 

Clinical Trials . Two contract-supported, randomized multicenter clinical 
trials on the treatment of diabetic retinopathy are in progress under BEP 
scientific management. These are the Early Treatment Diabetic Retinopathy 
Study (ETDRS) and the Diabetic Retinopathy Vitrectomy Study (DRVS). 

The ETDRS was designed to provide a better understanding of the best time 
to use photocoagulation in the course of diabetic retinopathy. Patients with 
macular edema, preprol if erative retinopathy, and mild or moderate prolifer- 
ative retinopathy are being studied. Three forms of photocoagulation 
treatment, ranging from restricted focal treatment to complete panretinal 



photocoagulation, are being compared with no photocoagulation. In addition, 
the study is evaluating the effect of daily administration of aspirin, in a 
comparison with placebo controls, on the incidence of microvascular and 
macrovascular complications. The study is also investigating factors 
associated with the progression of disease. Recruitment was completed in 
March 1985 with the enrollment of 3,928 patients. In December 1985 the study 
reported that focal photocoagulation of clinically significant diabetic 
macular edema substantially reduced the risk of visual loss. It was further 
reported that focal treatment increases the chances of visual improvement, 
decreases the frequency of persistent macular edema, and causes only minor 
visual field losses. Analysis files containing all pre-randomization data 
have been prepared by the coordinating center. Writing teams of clinical 
investigators have been formed and are working with these files. Two 
additional manuscripts have been published in the last year. Drs. Lloyd 
Aiello and Frederick L. Ferris, III, serve as Co-Chairmen for the ETDRS, and 
Dr. Richard L. Mowery serves as Project Officer. 

The DRVS has recruited a group of patients having a total of 997 eyes 
eligible for the study: 616 eyes with vision reduced by hemorrhage into the 
vitreous (group H) and 381 eyes still having useful vision but with serious 
risk of complications that often lead to retinal detachment (group NR). 
Follow-up of the NR group ended in mid-1988. Two publications have now 
appeared from this study. The most recent described the two-year status of 
eyes in the hemorrhage group. Its most important finding was a higher 
percentage with good vision in eyes assigned to early vitrectomy. This 
treatment advantage was particularly apparent for juvenile onset diabetics, 
possibly because of more active retinopathy. A manuscript on three-year 
results in group NR eyes has been submitted for publication. 

Dr. Sperduto was active in the scientific management of a grant-supported 
clinical trial, the Prospective Evaluation of Radial Keratotomy Study (PERK), 
which is designed to evaluate a surgical procedure — radial keratotomy — to 
correct myopia. Three-year results of the study were published in October 
1987. 

The Clinical Trials Branch implemented the Krypton-Argon Regression of 
Neovascularization Study (KARNS) in three pilot clinics in December 1983 to 
test the examination procedures and data collection forms. The major 
objective of this randomized clinical trial is to compare krypton laser to 
argon laser panretinal photocoagulation for treating neovascularization on the 
optic nerve head caused by diabetic retinopathy. The pilot phase was 
successfully completed in June 1984 and 29 new clinics were enrolled in KARNS 
starting in August 1984. As of July 1, 1988, a total of 849 patients had been 
randomized. This study is unique for the National Eye Institute since the 
functions for both the coordinating center and the fundus photography reading 
center are being handled by staff of the Clinical Trials Branch. Another 
feature of this multicenter trial is that the participating clinics receive no 
financial reimbursement from the National Eye Institute for their partici- 
pation. Dr. Ferris and Dr. Chew help direct this study. Dr. Mowery serves as 
Director of the Coordinating Center. 

The Clinical Trials Branch is also participating in the Diabetic Macular 
Edema Study. This Study is designed to compare two different treatment 
techniques for diabetic macular edema. The first is the treatment technique 



demonstrated to be effective in the Early Treatment Diabetic Retinopathy Study 
and the second is a "grid" technique that was shown to be effective in a small 
clinical trial and which has become popular. The Study has eight clinics that 
have enrolled 155 patients to answer questions raised by the study section 
review of the initial grant submission. Resubmission of the grant application 
will occur in late 1988 when early follow-up data on these randomized patients 
is available. Drs . Chew and Ferris are involved in this project. 

Dr. Seigel assisted Dr. Robert Turner in preparation of a grant 
application for a clinical trial on diabetes control and retinopathy, in 
Oxford, England. The study has been approved and funded. Dr. Seigel serves 
as the Institute's representative to the study's Data Monitoring Committee, 
which had its first meeting in the Spring of 1988. 

Dr. Seigel is serving as Project Officer for a randomized trial of 
sorbinil, a drug manufactured by Pfizer Laboratories. The drug is an aldose 
reductase inhibitor and has potential for preventing or retarding diabetic 
neuropathy and retinopathy. The NEI is providing scientific leadership for 
this multiclinic trial, which is funded by Pfizer. Approximately 500 patients 
have been randomized to treatment and follow-up which ended in mid-1988. 

Kathryn Chantry has been appointed as Project Officer for the statistical 
contract with the Orkand Corporation. Daniel Seigel serves as alternate 
Project Officer. The Orkand Corporation provides computer support to several 
of our scientific projects. 

Epidemiology . Patients continue to be recruited for a multicenter case- 
control study of selected retinal diseases. The study is attempting to 
identify possible risk factors for branch retinal vein occlusion, central 
retinal vein occlusion, idiopathic macular holes, rhegmatogenous retinal 
detachment, and exudative macular degeneration. Cardiovascular risk factors 
are of special interest. Dr. Sperduto and Dr. Seigel are Co-Chairmen of the 
study. Dr. Mowery serves as Project Officer. Ms. Rita Hiller, Dr. Chew, and 
Dr. Tamboli are members of the Project Team. 

Clinical reexamination of the original Framingham Eye Study participants 
for lens and macular changes, and photographic evaluation for macular 
degeneration, is proceeding under research contracts with Epistat Associates 
and the University of Wisconsin. The examinations will be completed in FY 88, 
and a photograph grading system will be completed by March 1989. Dr. Milton 
is Project Officer and Dr. Ferris is Alternate Project Officer for this Study. 

Dr. Tamboli, Dr. Sperduto, and Mr. Marvin Podgor are using the SEER 
(Surveillance, Epidemiology, and End Result) data to study the incidence of 
and survival rates for retinoblastoma. 

Dr. Sperduto is a Co-Principal Investigator in a joint Indo-American 
case-control study of aging-related cataracts. The study, which is being 
conducted in New Delhi, India, completed patient recruitment in December 
1987. An Investigators' meeting attended by Drs. Sperduto and Milton and Mr. 
Podgor was held in Delhi in February 1988. A preliminary review of the data 
was conducted at the meeting and a more complete analysis of the data is now 
in progress. 



Dr. Sperduto is the Project Officer for the joint Italian-American Case- 
ControL Study of Senile Cataract. The study is designed to identify risk. 
factors for aging-related cataracts. Recruitment of patients into the study 
began in the Spring of 1987 and is scheduled for completion in the Spring of 
1989. Because the study design is similar to that of studies being conducted 
in Boston, Massachusetts, and New Delhi, India, comparison of results among 
studies should be possible. 

Dr. Sperduto, Dr. Milton, and Dr. Mowery collaborated with Chinese 
investigators from the Peking Union Medical College in conducting a prevalence 
survey of cataract in Tibet. The study demonstrated a 60% increase in the 
prevalence of cataract in Tibet compared with the prevalence in a suburb of 
Beijing. A poster describing the study was presented at the ARVO meeting. 

Dr. Sperduto was the coauthor of papers that described and evaluated 
systems to quantify cataracts in vivo. The systems use photographic 
transparencies as standards to grade lens changes at the slit lamp or in color 
photographs. The systems were found to be highly reproducible and of 
potential value in cross-sectional and longitudinal studies. 

Mr. Podgor and Dr. Sperduto have collaborated with Dr. William Kannel 
(Boston University) and Dr. Gary Cassel (Wilmer Institute) in an investigation 
of possible associations of lens changes and the incidence of cardiovascular 
events among diabetics, using Framingham Eye Study data and follow-up data 
from the Framingham Heart Study. A manuscript is in preparation. 

Ms. Hiller, Dr. Sperduto, Mr. Podgor, Dr. Ferris, and Dr. Wilson 
collaborated on a paper that used data from the Framingham Heart Study and the 
Framingham Eye Study to examine the association between diabetic retinopathy 
and the occurence of cardiovascular events (coronary heart disease, inter- 
mittent claudication, congestive heart failure, or stroke) in Type II dia- 
betics. A paper is in press in the American Journal of Epidemiology. 

Dr. Sperduto, Mr. Podgor, and Ms. Hiller have collaborated with Drs. 
Manuel Datiles, Kayoko Kashima, and Paul Edwards of the Clinical Branch on the 
quantification of measurement error in grading retroillumination photographs 
of posterior subcapsular opacities. A manuscript is in preparation. 

Dr. Milton is collaborating with Dr. David Felson, multipurpose Arthritis 
Center, Boston City Hospital, in use of Framingham Eye Study data for a study 
of visual impairment and hip fracture. A presentation was made at the 
American Federation Clinical Research, and a manuscript is being submitted for 
publication. 

Dr. Sperduto continued his collaboration with Dr. M. Christina Leske in 
conducting a grant-funded, case-control study of aging-related cataracts. The 
Boston-based study seeks to identify risk factors for specific types of aging- 
related cataracts and to develop standardized techniques for diagnosing 
cataracts. Recruitment for the study will be completed in December 1988. 

Dr. Mowery serves as the Project Director for an operations research 
project being conducted at Aravind Eye Hospital in Madurai, India. The 
purpose of the three-year study is to investigate which of four approaches is 



the most effective in recruiting people to an eye clinic for cataract surgery 
and which method is most cost effective. Preliminary results were presented at 
the 1988 AfiVO meeting. Dr. Mowery serves on both the Executive and Steering 
Committees for this project. 

The Helen Keller International (HKI) supported "cataract free zone" 
projects in Peru and Brazil began in December 1986 and were completed in June 
1987. Dr. Mowery was involved in monitoring the progress of these studies and 
reviewing the progress reports for HKI. He visited both Peru and Brazil in 
1987 and 1988 to work with the investigators in preparing drafts of their 
final reports for presentations in November 1987 and posters that were 
presented in May 1988 at the ARVO meeting. 



Education ; 

Dr. Kupfer and Dr. Mowery presented lectures at the 1988 meeting of the 
American Association of Pediatric Ophthalmologists on clinical trials and 
epidemiologic methods for doing clinical research. 

During 1987-8, Dr. Ferris and Dr. Chew taught courses at the American 
Academy of Ophthalmology and several university centers on diabetic 
retinopathy and macular degeneration. 

Dr. Carl Kupfer, Dr. Ferris, Dr. Seigel, Dr. Sperduto and Dr. Milton 
participated as faculty in the eighth of a series of annual courses on 
epidemiologic and biostatistical approaches to clinical vision research. 
Along with four university colleagues and a former BEP associate director, 
Drs. Theodore Colton, Matthew Davis, Charles Hennekens, Lawrence Rand and Fred 
Ederer, they presented a three-day course in Sarasota, Florida for clinical 
investigators just before the 1988 ARVO annual meeting. The course was 
attended by about eighty people from academic institutions and was well 
received. Plans are under way for a ninth course in 1989. 

Dr. Ferris collaborated with the American Academy of Ophthalmology to 
prepare a videotape summarizing the clinical implications of the results of 
the Early Treatment Diabetic Retinopathy on the treatment of diabetic macular 
edema. 

Drs. Seigel and Sperduto supervised the training program for three staff 
fellows from China: Drs. Jingjing Xu, Lizong Hu, and Li-Qi Tang. 

Collaboration and Consultation 

Dr. Ferris is a member of the Data, Safety, and Quality Review Board for 
the Diabetes Control and Complications Trial, National Institute of Arthritis, 
Diabetes, and Digestive and Kidney Diseases. He is also a member of the DRVS 
Data Monitoring Committee, and Data and Safety Monitoring Committee of the 
grant-supported Collaborative Ocular Melanoma Study. 

Dr. Milton provided biostatistical and administrative support through 
consultation and review for several international projects in ophthalmic 
research, including the US-Indo Science and Technology Initiative programs. 



Dr. Mowery served on Che NCI's Intramural and Administrative Support 
Contract Review Committee. He also serves as the Project Director for a data 
management support contract that serves the needs of all NEI staff. 

Mr. Podgor consulted with Dr. Griffin Rodgers, NIDDK on hypertension in 
sickle ceil disease. 

Mr. Podgor consulted with Deborah Street, Johns Hopkins University, on 
sample size estimation for an AIDS case-control study. 

Mr. Podgor consulted with Dr. Monique Roy, Clinical Branch, NEI on color 
vision in normal volunteers. 

Dr. Seigel served on an NIH Director's panel on hiring and promotions for 
epidemiologists and statisticians. 

Dr. Sperduto collaborated with Dr. Datiles of the NEI ' s Clinical Branch 
on the use of photographic techniques to document the presence and progression 
of lens opacities. A study was completed that estimated the measurement error 
and its effect on sample size requirements in clinical studies when two 
measurement systems were used to quantitate the size of posterior subcapsular 
opacities as seen in retroillumination photographs. 

Dr. Sperduto served as an ophthalmic consultant to NEI ' s Office of 
Planning and Reporting. 

Dr. Sperduto assisted in preparing a report on long-range planning of the 
National Eye Institute's cataract program. The report will be used as the 
basis for the Cataract Program Section of the next report of the National 
Advisory Eye Council for the period 1990-1992. 

Dr. Freidlin consulted with Dr. Datiles on statistical methods for 
comparing endothelial cells of diabetic and non-diabetic patients. 

Dr. Freidlin consulted with Dr. Edwards on the analysis of the computer 
classification of different types of cataracts. She will be acknowledged in 
the paper. 

Dr. Freidlin collaborated with Dr. Roy on the early results of Aging- 
Related Macular Degeneration (AMD) Study. 

Dr. Freidlin consulted with Dr. Kaiser-Kupfer on lens opacities in 
patients with bilateral acoustic neurofibromatosis. A manuscript is being 
prepared. 



Professional Activities ; 

Dr. Milton is a member of the Management Committee for "Current Index to 
Statistics," representing the American Statistical Association. 

Dr. Mowery served as Chairman of the Membership Committee of the Society 
for Clinical Trials. 

Dr. Seigei served on the Editorial Board of two journals: the Archives of 
Ophthalmology and Statistics in Medicine. 

Dr. Sperduto is a member of the Data Monitoring Committee for a grant- 
supported clinical trial on retinitis pigmentosa. 

Dr. Sperduto is a member of the Data Monitoring Committee for the 
Prospective Evaluation of Radial Keratotomy Study. 

Dr. Sperduto serves on the Advisory Committee for the Wisconsin 
Epidemiologic Study. 



Presentations 

Dr. Ferris was an invited speaker for a symposium on data monitoring at 
the Clinical Trials Society meeting. 

Dr. Mowery presented a lecture at the Eighth Annual Meeting of the 
Society for Clinical Trials on quality assurance issues in clinical trials. 

Dr. Seigei collaborated with Dr. A. Hillis in writing a talk, on surrogate 
statistics in eye research, given at the Biometrics Society annual meeting. 
It is in press in Statistics in Medicine. 

Dr. Seigei presented a lecture to ophthalmology residents at Howard 
University on the principles of clinical research. 

Drs. Seigei and Milton reported on their Monte Carlo analyses of grading 
systems for lens opacities, making presentations at Johns Hopkins School of 
Medicine and at NIH. A manuscript summarizing the results has been submitted 
for publication. 



Publications; 

1. Aiello LN, Ferris FL. Photocoagulation for diabetic macular edema 
(Letter to the Editor). Arch OphthaLmoL 1987;105 : 1163 . 

2. Chylack. LT, Leske MC, Sperduto RD, Khu P, et al. Lens opacities 
classification system. Arch Ophthalmol 1988;106:330-4. 

3. Datiles MB, Edward PA, Kaiser-Kupf er MI, McCain L, Podgor M. A 
comparative study between the PAM and the laser interferometer in 
cataracts. Graefe's Arch Clin Exp Ophthalmol 1987;225:457-60. 

4. Datiles M, Podgor M, Edwards P. Reproducibility study on the early 
cataract detector (Kowa ECD 2000). Ophthalmic Surg (in press). 

5. Early Treatment Diabetic Retinopathy Study Research Group. Techniques for 
scatter and local photocoagulation treatment of diabetic retinopathy: 
ETDRS Report No. 3. International Ophthalmol Clinics. 1987 ;27(4) :254- 
64. Little, Brown & Co., Boston. 

6. Early Treatment Diabetic Retinopathy Study (ETDRS) Research Group. 
Photocoagulation for diabetic macular edema. ETDRS Report No. 4. 
International Ophthalmol Clinics. 1988;28:265-72. Little, Brown & Co., 
Boston. 

7. Hiller R, Sperduto RD, Podgor MJ, Ferris FL, Wilson PWF. Diabetic 
retinopathy and cardiovascular disease in type II diabetics: the 
Framingham Heart Study and the Framinghara Eye Study. Am J Epidemiol 
1988;128:402-9. 

8. Hillis A, Seigel D. Surrogate observations in ophthalmologic studies. 
Statistics in Medicine (in press). 

9. Kaufman SC, Ferris FL, Swartz M, DRS Research Group. Diabetic 
Retinopathy Report No. 11. Arch Ophthalmol 1987; 105 :8079. 

10. Leske MC, Chylack. LT, Sperduto RD, Khu P, et al. Evaluation of a lens 
opacities classification system. Arch Ophthalmol 1988;106:327-9. 

11. Leske MD, Chylack LT, Sperduto R, Pennett M and McCarthy D. Progress 
toward developing a cataract classification system. In: "Developments in 
Ophthalmology," S. Karger Publisher, Basel/Switz 1987, vol 15, pp 9-15. 

12. Milton RC, Mohan M, Sperduto RD. Indo-US Case-control study of senile 
cataract design and development, in Straub (ed): "Developments in 
Ophthalmology." S. Karger Publisher, Basel/Switz 1987, vol 15, pp 92-98. 

13. Milton RC, Reddy V, and Naidu AN. Mild vitamin A deficiency and 
childhood morbidity - an Indian experience. Am J Clin Nutr 1987;46:827- 
9. 

14. Nussenblatt RB, Kaufman SC, Palestine AG, Davis MD, Ferris FL. Macular 
Thickening and Visual Acuity. Measurement in Patients with Cystoid 
Macular Edema. Opthalmol 1987 ;94(9) : 1134-8 . 



15. Rosner B, MilCon RC. Significance testing for correlated binary outcome 
data. Biometrics 1988;44:505-12. 

16. Roy MS, Podgor M J , Rick ME. Plasma f ibrinopeptide A, b-thromboglobulin, 
and platelet factor 4 in diabetic retinopathy. Invest Ophthalmol Vis Sci 
1988;29:856-60. 

17. Roy MS, Podgor MJ, Bungay P, Grunberger G, Carl J, Ellis D. Posterior 
vitreous f lourophotometry in diabetic patients with minimal or no 
retinopathy. Retina 1987 ; 7 : 170^6 . 

18. Seigel D. Designs for clinical research. Arch Ophthalmol. Dec 
1987;105:1647-9. 



CONTRACT NARRATIVE 

Thirteen Clinical Centers; a Coordinating Center at the University of 
Minnesota, Minneapolis, Minnesota; and a Fundus Photograph Reading Center at 
the University of Wisconsin, Department of Ophthalmology, Madison, Wisconsin. 

Title ; Diabetic Retinopathy Vitrectomy Study (DRVS) 

Principal Investigators ; Matthew D. Davis, M.D. (Study Chairman) 

Daniel Seigel, Sc.D. (Project Officer) 

Current Fund Allocation ; $144,966 (estimate) FY 1988 (EY 5 2148, EY 5 2147) 

Objectives ; The DRVS is a multicenter clinical trial to: 

1. Evaluate vitrectomy performed in the first six months after severe 
vitreous hemorrhage secondary to diabetic retinopathy compared to the 
more usual practice of waiting twelve months after vitreous hemorrhage 
to remove the vitreous (group H). 

2. Evaluate vitrectomy in eyes with good vision but with severe 
proliferative retinopathy and poor prognosis before vision is lost 
through hemorrhage or retinal detachment (group NR). 

3. Study the natural history of severe proliferative diabetic 
retinopathy. 

Major Findings ; The first report of results for eyes with severe vitreous 
hemorrhage was published in the November 1985 issue of the Archives of 
Ophthalmology. Over six hundred eyes with recent severe diabetic vitreous 
hemorrhage were randomly assigned to either early vitrectomy or deferral of 
vitrectomy for one year. After two years of follow-up, 25% of the early 
vitrectomy group had visual acuity of 10/20 or better compared with 15% in the 
deferral group. 

Significance to Biomedical Research and the Program of the Institute ; 
Diabetic retinopathy is one of four major causes of adult blindness and 
differs from the other three (macular degeneration, glaucoma, cataract) in 
that it generally affects a younger population. Vitrectomy has the 
theoretical potential of removing the "scaffolding" on which abnormal new 
vessels can develop, fibrous tissue can form, and retinal detachment can 
occur. It is important to determine when such intervention is most likely to 
deter this process and reduce the incidence of loss of vision. 

Proposed Course ; Follow-up has been completed. A manuscript has been 
submitted on 3-year results in the NR series. In 1988, 4-year results in the 
hemorrhage series will be analyzed. 

NEI Research Program ; Retinal and Choroidal Diseases 



Publications ! 

The Diabetic Retinopathy Vitrectomy Research Group. Two-year course of 
visual acuity in severe proliferative diabetic retinopathy with 
conventional management. DRVS Report No. 1. Ophthalmology, 92:492-502, 
1985. 

The Diabetic Retinopathy Vitrectomy Study Group. Early vitrectomy for 
severe vitreous hemorrhage in diabetic retinopathy. Two year results of a 
randomized trial. Diabetic Retinopathy Vitrectomy Study Report Number 
2. Arch Ophthalmol 103:1644-1652, 1985. 



CONTRACT NARRATIVE 

Twenty-chree Clinical Centers; a Coordinating Center at Maryland Medical 
Research Institute; a Fundus Photograph Reading Center at the University of 
Wisconsin, Department of Ophthalmology, Madison; a Central Laboratory at the 
Centers for Disease Control, Atlanta, Georgia; and an Electrocardiogram 
Reading Center at the University of Minnesota, Minneapolis, Minnesota. 

Title ; Early Treatment Diabetic Retinopathy Study (ETDRS) 

Principal Investigators ; Dr. Lloyd Aiello (Co-Chairman) 

Dr. Frederick. L. Ferris, III (Co-Chairman) 
Dr. Richard L. Mowery (Project Officer) 

Current Fund Allocation ; $5,696,686 (estimated) for FY 1988 

Objectives ; The Early Treatment Diabetic Retinopathy Study (ETDRS) is a 
multicenter randomized clinical trial, Che main goals of which are; 

1. To determine whether treatment of early stages of proliferative and 
nonproliferative diabetic retinopathy, with or without macular edema, 
by aspirin and/or prompt photocoagulation is effective in decreasing 
the rate of development of known retinopathy risk factors and/or the 
development of severe visual loss when compared to placebo or deferred 
photocoagulation. 

2. To help determine the best time to initiate photocoagulation treatment 
in diabetic retinopathy. 

3. To monitor closely the effects of diabetes mellitus and/or of 
photocoagulation on visual function. 

4. To produce natural history data that can be used to develop (identify 
risk factors) and test etiologic hypotheses in diabetic retinopathy. 

Major Findings ; From April 1980 to March 1985, the ETDRS research group 
enrolled 3,928 diabetic patients with early proliferative retinopathy, 
moderate to severe nonproliferative retinopathy, and/or diabetic macular edema 
in each eye. In December 1985, the research group published a report that 
focal photocoagulation of "clinically significant" diabetic macular edema 
substantially reduces the risk of visual loss. Focal treatment also increases 
the chance of visual improvement, decreases the frequency of persistent 
macular edema, and causes only minor visual field losses. 

Significance to Biomedical Research and the Program of the Institute ; The 
National Eye Institute regards fostering careful evaluation of new and widely 
used ophthalmic treatments as an essential element in its mission. This study 
represents an extension of the Institute's interest in preventing visual 
impairment of patients with diabetes. 

Proposed Course ; The study will end patient follow-up in July 1989 and 
prepare reports at that time on study results. 



NEI Research Program ; Retinal and Choroidal Diseases 

Publications ; 

Early Treatment Diabetic Retinopathy Study Research Group: 
Photocoagulation tor Diabetic Macular Edema. Arch Ophthalmol 103:1796, 
1985. 

Early Treatment Diabetic Retinopathy Study Research Group: 
Photocoagulation Therapy for Diabetic Eye Disease. JAMA 254:3086, 1985. 

Early Treatment Diabetic Retinopathy Study Research Group: Treatment 
Techniques and Clinical Guidelines for Photocoagulation of Diabetic 
Macular Edema, Report Number 2. Ophthalmology, 94:761-774, 1987. 

Early Treatment Diabetic Retinopathy Study Research Group: Techniques for 
Scatter and Local Photocoagulation Treatment of Diabetic Retinopathy : 
Early Treatment Diabetic Retinopathy Study Report No. 3. Internat Ophthal 
Clinics, 1987;27(4):254-64. Little, Brown & Co. Boston. 

Early Treatment Diabetic Retinopathy Study (ETDRS) Research Group: 
Photocoagulation for Diabetic Macular Edema. ETDRS Report No. 4. 
Internat Ophthalmol Clinics, 1988;28:265-72. Little, Brown & Co. Boston. 

Ferris FL and Aiello LM: Photocoagulation for Diabetic Macular Edema. 
Letter to the Editor. In press. 



CONTRACT NARRATIVE 

Five Clinical Centers; a Central Laboratory at National Health Laboratories, 
Vienna, Virginia; a Nutrition Biochemistry Laboratory at Centers for Disease 
Control, Atlanta, Georgia; an Electrocardiogram Reading Center at the 
University of Minnesota, iMinneapolis , Minnesota; a Data Management Group at 
Rockville, Maryland. 

Title ; Eye Disorders Case Control Study (EDCCS) 

Principal Investigators ; Dr. Daniel Seigel (Co-Chairman) 

Dr. Robert Sperduto (Co-Chairman) 
Dr. Richard Mowery (Project Director) 

Current Fund Allocation ; $801,075 (estimated) for FY 1988 

Objectives ; The goal of the Eye Disorders Case Control Study is to evaluate 
the role of potential risk factors for a number of disorders of the eye for 
which adequate epidemiologic data are now lacking. Secondary objectives of 
the study are to evaluate grading systems, particularly for hypertensive and 
arteriosclerotic changes in the retina. 

Major Findings ; Pilot testing at each of the four clinical centers was done 
between February-May 1986 based on the Manual of Operations designed by the 
NEI staff and the clinic staffs. Each clinic recruited at least five 
patients. The main study began in June 1986. Over 700 cases and 400 controls 
have been recruited. Wilmer Eye Clinic at Johns Hopkins Hospital has been 
added as a fifth clinic. 

Significance to Biomedical and the Program of the Institute ; In the 1983 
Report of the National Advisory Eye Council, a need was identified for 
"epidemiologic studies on various types of retinal vascular disease with 
particular view to isolating causative factors." In recent years, careful 
epidemiologic studies have been initiated for diabetic retinopathy, aging- 
related macular degeneration and ocular melanoma. However, for various forms 
of retinal artery and vein occlusions and rhegmatogenous retinal detachments, 
high quality epidemiologic data are lacking. In particular, cardiovascular 
risk factors appear to be associated with these disorders. This study 
represents an extension of the Institute's interest in identifying risk 
factors associated with retinal diseases. 

Proposed Course ; The five clinical centers will continue to recruit both 
cases and controls for four years. As soon as at least two hundred cases have 
been examined in any one disease group, analyses will begin. 

NEI Research Program ; Retinal and Choroidal Diseases 

Publications: None 



CONTRACT NARRATIVE 

A Fundus Photograph Reading Center at the Department of Ophthalmology, 
University of Wisconsin, Madison 

Title : Reading Center for Framinghara Eye Study Photographs 

Principal Investigators : Matthew D. Davis, M.D. (Principal Investigator) 

Roy C. Milton, Ph.D. (Project Officer) 
Frederick L. Ferris, III, M.D. (Alternate Project 
Officer) 

Current Fund Allocation : $75,838 (estimated) FY 1987 for EY 62116 

Objectives : To develop a classification system for aging-related macular 
disease and to provide an evaluation of that disease using the fundus 
photographs from the 1973-1975 Eye Study and the fundus photographs to be 
taken in the 1986-1988 Framingham Eye Study. 

Major Findings : This study began in June 1986 and is a companion study to The 
Ocular Re-examination of Framingham Eye Study Subjects. Development of the 
classification system is complete and application is ongoing. 

Significance to Biomedical Research and the Program of the Institute : Aging- 
related macular degeneration is a major cause of blindness. Incidence rates 
for this disease are not available, and the natural history is largely 
unknown. The data from this study on incidence, progression, and association 
with other variables could lead to an increased understanding of this aging- 
related ocular disease and possibly to the development of measures to prevent 
or delay its onset. This study is consistent with the Institute's interest in 
epidemiologic research and in alleviation of the human and economic burden of 
eye disease. 

Proposed Course : The classification scheme for aging-related macular degener- 
ation will be applied to approximately 1500 fundus photographs from the 1973- 
75 study and 1000 fundus photographs to be taken during 1986-88. Quality 
control and monitoring of evaluation methods and results will be ongoing. 
Photograph classification will be completed by March 1989. 

NEI Research Program : Retinal and Choroidal Diseases 

Publications: None. 



«' 



CONTRACT NARRATIVE 

A clinical Examination Center at Framingham, Massachussects , operated by 
Epistat Associates, Incorporated. 

Title : Ocular Re-examination of Framingham Eye Study Subjects 

Principal Investigators ; Theodore Colton, Sc.D. (Principal Investigator) 

Lawrence Rand, M.D. (Co-Investigator) 
Roy C. Milton, Ph.D. (Project Officer) 
Frederick L. Ferris, III, M.D. (Alternate Project 
Officer) 

Current Fund Allocation ; $240,266 (estimated) for FY 1988 EY 2105 

Objectives ; This re-examination of Framingham Eye Study subjects, first seen 
in 1973-1975, is an epidemiologic study of aging-related macular degeneration 
and cataract to determine their incidence, to describe their natural history, 
and to identify associations between their presence or progression and 
variables in the Framingham Heart Study, whose values were determined before 
development or progression of these diseases. 

Major Findings ; This study began in January 1986. Study procedures have been 
developed, equipment has been purchased and installed, and examination staff 
have been hired and trained. Examination of study subjects began in August 
1986. About 1000 subjects will be examined during the study, and examinations 
will be completed by December 1988. 

Significance to Biomedical Research and the Program of the Institute ; Aging- 
related macular degeneration and cataracts are major causes of blindness in 
the United States, accounting for thirteen and nine percent of all blindness, 
respectively. Incidence rates for these diseases are not now available, and 
their natural history is largely unknown. The data from this study on incid- 
ence, progression, and association with other variables could lead to an 
increased understanding of these aging-related ocular disease and possibly to 
the development of measures to prevent or delay their onset. This study is 
consistent with the Institute's interest in epidemiologic research and in 
alleviation of the human and economic burden of eye disease. 

Proposed Course ; Examination of an estimated 1000 study subjects will be 
completed by December 1988. Quality control procedures and monitoring of data 
is ongoing. 

NEI Research Program ; Retinal and Choroidal Diseases 

Publications; None 



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