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 0 (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 Researcheris
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 excaeH 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
0 (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 umtjaiu 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 oftonmi 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
0
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 nn743-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>» oum&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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