Cytokeratin subtyping to distinguish reactive and neoplastic RPE cells.
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Biomarkers, Tumor↗
Biomedical subjects
P S Hiscott
Publications and source records attributed to P S Hiscott.
Microarray comparative genomic hybridisation analysis of intraocular uveal melanomas identifies distinctive imbalances associated with loss of chromosome 3.
Defining regions of genomic imbalance can identify genes involved in tumour development. Conventional cytogenetics has identified several nonrandom copy number alterations (CNA) in uveal melanomas (UVM), which include monosomy 3, chromosome 6 abnormalities and gain of 8q. To gain further insight into the CNAs and define the regions involved more precisely we analysed 18 primary UVMs using 1 Mb BAC microarray comparative genomic hybridisation (CGH). Our analysis showed that the most common genomic imbalances were 8q gain (78%), 6p gain (67%) and monosomy 3 (56%). Two distinct CGH profiles could be delineated on the basis of the chromosome 3 status. The most common genetic changes in monosomy 3 tumours, in our study, were gain of 8q11.21-q24.3, 6p25.1-p21.2, 21q21.2-q21.3 and 21q22.13-q22.3 and loss of 1p36.33-p34.3, 1p31.1-p21.2, 6q16.2-q25.3 and 8p23.3-p11.23. In contrast, disomy 3 tumours showed recurrent gains of only 6p25.3-p22.3 and 8q23.2-q24.3. Our approach allowed definition of the smallest overlapping regions of imbalance, which may be important in the development of UVM.
Adult↗
Immunophenotypic shift in a case of mycosis fungoides with vitreous invasion.
The case of an 82-year-old man who developed intraocular extension from mycosis fungoides, a cutaneous T-cell lymphoma, is presented. The patient died soon after intraocular involvement occurred. Immunohistochemistry of a skin biopsy, taken early in the course of the disease, disclosed a predominance of T cells with a helper/inducer phenotype (CD4(+)). However, an intraocular infiltrate obtained 7 years later contained mostly T cells with a suppressor/cytotoxic phenotype (CD8(+)). The occurrence of ocular invasion, the change in immunophenotype, and the predominant proliferation of CD8(+) lymphocytes may have been related to the poor outcome in this patient.
Aged↗
Biomaterials used in the posterior segment of the eye.
The treatment of posterior segment eye disease and related conditions has improved greatly in recent years with the advent of new therapies, materials and devices. Vitreoretinal conditions, however, remain significant causes of blindness in the developed world. Biomaterials play a major role in the treatment of many of these disorders and the success rate of vitreoretinal surgery, especially in the repair of retinal detachment and related conditions, would increase with the introduction of new and improved materials. This review, which focuses on disorders that feature retinal detachment, briefly describes the anatomy of the eye and the nature and treatment of posterior segment eye disorders. The roles, required properties and suitability of the materials used in vitreoretinal surgery as scleral buckles, tamponade agents or drug delivery devices, are reviewed. Experimental approaches are discussed, along with the methods used for their evaluation, and future directions for biomaterial research in the posterior segment of the eye are considered.
Biocompatible Materials↗
Synthesis of osteonectin by human retinal pigment epithelial cells is modulated by cell density.
PURPOSE: To determine whether human retinal pigment epithelial (HRPE) cells are able to synthesize the antiadhesive protein osteonectin, also known as secreted protein, acidic and rich in cysteine (SPARC). Additionally, because locally produced SPARC may modulate cellular behavior during tissue repair, to ascertain whether HRPE SPARC production and HRPE proliferation, migration, and/or differentiation are associated, in a simple HRPE wound-healing model. METHODS: Immunohistochemical and Western blot analyses of SPARC protein expression by low- and high-density cultured HRPE cells were undertaken. Total RNA extracted from cultures was studied by reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis. Western and Northern blot analyses were evaluated by densitometry. Experiments were repeated with HRPE cells cultured in the presence of 1, 10, or 100 microM of the differentiating agents butyric acid (BA) and retinoic acid (RA). RESULTS: HRPE cell cultures exhibited SPARC immunoreactivity. Western blot analysis of cell lysates and conditioned media showed a 43-kDa protein. RT-PCR and Northern blot analysis confirmed the presence of SPARC mRNA (with transcripts at 2.2 and 3.0 kb). Protein and mRNA transcript band densitometry revealed a higher proportion of SPARC protein and mRNA in high-density HRPE cell culture than in low-density culture. Neither BA nor RA (at the concentrations assessed) had a significant effect on SPARC production by HRPE cells in high- or low-density culture. CONCLUSIONS: HRPE can synthesize SPARC. Although the findings do not support an invariable association between SPARC production by HRPE and HRPE proliferation, migration, or differentiation, they demonstrate that synthesis of SPARC by HRPE is modulated by cell density.
Blotting, Northern↗
E coli panophthalmitis with orbital cellulitis.
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Aged↗
Cytokeratins and retinal epithelial cell behaviour.
The expression of cytokeratins 18 and 19 by human retinal pigment epithelial cells (HRPE) has been suspected of being associated with HRPE proliferation. We have investigated the involvement of these cytokeratin subtypes in the proliferative and migratory behaviour of cultured HRPE. Cell proliferation markers (bromodeoxyuridine and proliferating cell nuclear antigen) and the cytokeratins were identified using immunohistochemical techniques. In vitro, cytokeratins 18 and 19, as detected by the monoclonal antibodies RGE 53 and K4.62, were expressed in a subset of HRPE and this subset was significantly less likely to be proliferating. Micro-chemotaxis chambers were used to study migrating cells and immunohistochemical staining for cytokeratins 18 and 19 revealed that actively migrating cells always expressed these two cytokeratins, whereas stationary cells did not label for these cytokeratin subtypes. It was apparent that cytokeratins 18 and 19 were not markers of proliferation, but were involved in the mobility of HRPE in vitro. Cytokeratins 18 and 19 may be useful indicators of simple epithelial cell migration in tissues.
Actins↗
Sorsby's fundus dystrophy. A light and electron microscopic study.
The light and electron microscopic changes in both eyes of a patient with Sorsby's fundus dystrophy are reported. The most striking finding was a 30-microns thick deposit present within Bruch's membrane that stained positive for lipids. The deposit has several unique features when compared with those found in other retinal dystrophies and in aged eyes. In addition, there was gross loss of the outer retina, the retinal pigment epithelium was discontinuous, and there was atrophy of the choriocapillaris. These findings are discussed in relation to the published clinical observations of other members of this pedigree.
Choroid↗
Growth and contractility of cells from fibrocellular epiretinal membranes in primary tissue culture.
Cells grew out in tissue culture from 37 of 53 fibrocellular epiretinal membrane explants, those from membranes of less than four months' clinical duration growing more consistently, more rapidly, and more exuberantly than those from membranes of more than four months' clinical duration. Electron microscopy demonstrated prominent microfilaments in the cytoplasm of some of the cells, and immunostaining for actin showed great variations in the pattern and intensity of staining. An ATP contraction assay conducted on 44 detergent-extracted cells and 47 intact cells revealed in each case a 30% average reduction in cell area. Immunostaining for cytokeratins following the ATP assay showed no significant difference in cell contraction between epithelial cells and non-epithelial cells. Clinical duration of the membrane had no significant influence on the contractility of cells growing from the explant.
Actin Cytoskeleton↗
The role of inflammation in the development of epiretinal membranes.
Single or multiple (3) injections of "Shigella" endotoxin were administered into the rabbit vitreous body to investigate the development of epiretinal membranes following intraocular inflammation. The evaluation included clinical assessment of the resulting traction retinal detachments, together with histological, autoradiographic, immunohistochemical and ultrastructural studies. Traction retinal detachments were found beneath fibroglial epiretinal membranes (being more extensive in eyes which had received 3 endotoxin injections) in the vicinity of the medullary rays, while purely glial membranes occurred over attached peripheral retina. The primary change at the vitreoretinal interface was an elevation of the inner limiting lamina of the retina followed by the extension of glial cells onto the retinal surface. It is postulated that glial cells breach the inner limiting lamina as a sequel to inflammation involving the vitreoretinal interface and form a scaffold upon which fibroblast-like cells migrate.
Animals↗
Age-related cellular proliferation at the vitreoretinal juncture.
Clinical and pathological features of non-vascularised epiretinal membranes are reviewed with special attention to focal epimacular tractional lesions in the elderly. The role of immunohistochemistry in elucidating the nature of component cells of complex epiretinal membranes is emphasised. Clinicopathological correlation establishes the 'fibroglial membrane' as the causative lesion of age-related epimacular traction. The pathogenesis of this process is discussed, including relevant animal models, and chronic inflammation and ischaemia (rather than acute posterior vitreous detachment) are implicated. Vitrectomy and epimacular membrane peeling results in significant visual improvement in most patients.
Adult↗
Effect of melanin bleaching on immunoperoxidase, with reference to ocular tissues and lesions.
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Aged↗
The effects of injections of cultured fibroblasts into the rabbit vitreous.
A comparison was made between rabbit's eyes which had an intravitreal injection of either 400,000 autologous or homologous skin fibroblasts. The follow-up period was up to 6 months with autologous and 10 weeks with homologous cells. The evaluation involved the clinical assessment of membrane formation, retinal detachment and anterior segment inflammation. In addition, a detailed histological, electron microscopic and autoradiographic study was made of membrane development and changes in the surrounding tissue. Clinical evaluations revealed little difference between the two models. However, on the basis of macroscopic and microscopic examination, homologous cells produced more extensive detachments and more pronounced retinal gliosis. Plasma cells were evident in the choroid 6 weeks after homologous fibroblast injection, but they were not associated to any appreciable extent with the autologous model. Fibrovascular proliferation from around the optic nerve head made a major contribution to the formation of membranes in the experiments with homologous fibroblasts but not those using the autologous cells.
Animals↗
Natural history of fibrocellular epiretinal membranes: a quantitative, autoradiographic, and immunohistochemical study.
Quantitative, autoradiographic, or immunohistochemical light microscopy was conducted on 85 surgically excised epiretinal membranes to investigate the activities of component cells and the natural history of the extraretinal scars. Membranes of less than four months' clinical duration ('early' membranes) contained significantly more cells than 'late' membranes, while collagen was more abundant in the older specimens. No correlation was established between membrane opacification and either the cellularity or the collagen content of the membranes. Epiretinal membranes had some features in common with healing skin wounds, but the activities of cells in the epiretinal membranes were relatively protracted and disordered. Fibronectin was found to be a significant component of epiretinal membranes and therefore represents a target at which pharmacological intervention could be aimed.
Actins↗
Reparative epiretinal fibrosis after diabetic vitrectomy.
Closed microsurgery for the haemorrhagic and tractional sequelae of proliferative diabetic retinopathy is frequently successful in restoring and maintaining useful vision. Severe complications may arise, however, including the reproliferation of cellular membranes on the retinal surface with subsequent retinal distortion or redetachment. The clinical and histopathological features of such reparative epiretinal fibrosis are presented and methods of prevention are discussed.
Diabetic Retinopathy↗
Retinal and epiretinal glia--an immunohistochemical study.
Immunohistochemical techniques were used to examine the distribution of cells containing glial fibrillary acidic protein (GFAP) in normal and pathological human specimens, including 22 globes (13 of which contained epiretinal membranes 'in situ'), 16 surgically excised epiretinal membranes, and monolayers of cells obtained from five epiretinal membranes placed in tissue culture. The astrocytic cells of normal and pathological retinae stained with the glial-cell marker, but Müller cells were GFAP-negative in normal retinae at the antisera dilutions used. Müller cells did, however, stain in retinae from glaucomatous eyes and in eyes with prolonged retinal detachment. Electron microscopy did not reveal any obvious morphological difference between the intermediate filaments of normal (GFAP-negative) and GFAP-positive Müller cells. Ten of the 13 epiretinal membranes 'in situ', all 16 excised membranes, and three of the five monolayers contained glial cells. Purely glial membranes were not associated with retinal puckering or detachment, while all membranes causing tractional complications had a prominent fibrous, non-glial component. Our findings suggest that glial cells do not contribute significantly to the contractile forces generated by epiretinal membranes. They may, however, provide a scaffold on which other cells proliferate and contract and an anchorage by means of which tangential forces are transmitted into and through the retina.
Adolescent↗
Retinal pigment epithelial cells in epiretinal membranes: an immunohistochemical study.
Immunohistochemical techniques were used to identify cells containing cytokeratins in sections or tissue-culture monolayers from ocular (reference) tissues and also from 22 epiretinal membranes obtained during closed microsurgery for macular pucker or massive preretinal retraction. Results of cytokeratin immunostaining in reference tissues indicated that this is a valuable means of determining the contribution and distribution of epithelial cells in epiretinal membranes, and that the epithelial cells in the membranes were probably derived from the retinal pigment epithelium. Epithelial cells were identified in 17 of the 22 epiretinal membranes, but they did not usually constitute the predominant cell type. We concluded that the fibroblasts or fibroblast-like cells thought to be responsible for the contraction of epiretinal membranes are seldom of retinal pigment epithelial origin. Biomicroscopic pigmentation of a membrane was shown to be a poor guide to its epithelial cell population.
Animals↗
Epiretinal membranes in vitro.
Monolayers of cells were grown from epiretinal membranes using standard tissue culture techniques. The growing cells were filmed by time-lapse cinephotomicrography to record their locomotory characteristics and cell-to-cell interactions. The established layers of cells were examined in the scanning electron microscope. In addition, the cultured cells were studied immunohistochemically to identify specific cell types and the distribution of contractile elements (e.g. actin). The locomotory patterns, the topography and the immunohistochemistry of cells from epiretinal membranes were compared to the movement, appearance and staining characteristics of a wide range of normal cells in culture. From the growth characteristics in tissue culture, early membranes appeared more hypercellular than longer standing membranes. Cell outgrowth was established more quickly and was more widespread in the young epiretinal membranes than in older, more fibrous, specimens. The locomotory and immunohistochemical studies demonstrated that a wide variety of cell types may contribute to epiretinal membranes. A remarkably wide range of cell forms were observed in cultures of individual membranes. Epithelial and glial cells were found in many of the monolayers. However, they did not constitute the majority of cells in the cultures. The predominating cell types had the morphological and locomotory characteristics of fibroblasts or macrophages.
Animals↗