We concluded that TGF-beta up-regulates human RPE phagocytosis, but that this effect is counteracted by PKC activation. It is possible that this TGF-beta-induced effect is due, in part, to a negative modulation of the PKC-dependent pathway.

Retinal pigment epithelial (RPE) cells induced to express MHC class II (HLA-DR) by incubation with interferon gamma (IFN-gamma) were investigated for their ability to present a microbial superantigen to T lymphocytes. Superantigens bind to MHC class II antigens and appear to play a role in a number of infectious and autoimmune diseases through stimulation of large numbers of T cells. Primary cultures of human RPE cells treated with IFN-gamma for three days to induce HLA-DR expression bound staphylococcal enterotoxin E (SEE) via HLA-DR and presented SEE to T cells as measured by proliferation of purified peripheral blood T cells and IL-2 synthesis by the Jurkat T cell line. Untreated RPE cells were essentially ineffective as superantigen presenting cells. These results suggest that MHC class II expressing RPE cells could contribute to immune and inflammatory activity in the eye by presenting superantigens to T lymphocytes.

The present study indicates a relationship between ONH and finger blood flow in subjects with PVD. This might be an indirect sign of a disturbed autoregulation of ocular blood flow in PVD subjects.

Because fibronectin (FN) is known to be present in membranes in proliferative vitreoretinopathy, we sought to identify cytokines that regulate the release of FN by retinal pigment epithelial cells (RPE). Levels of FN in the supernatant of cultured human RPE cells were quantified with an ELISA, after which the cells were stimulated with human recombinant cytokines, interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF alpha), interleukin-6 (IL-6), interferon gamma (IFN-gamma), transforming growth factor beta 1 and 2 (TGF-beta), and phorbol myristate acetate (PMA). Protein kinase C (PKC) was blocked by 2 nM calphostin C or 1 mM staurosporine. RPE cells released FN into the supernatant constitutively. TGF-beta 1 and TGF-beta 2 upregulated the FN release in a dose- and time-dependent manner. The other cytokines tested were without effect. In combination, IFN-gamma and IL-1 beta reduced the effect of TGF-beta. PMA, which is a PKC activator, also increased the release of FN in a dose-dependent manner. Blocking of PKC with specific inhibitors abolished the effects of TGF-beta and PMA. The results show that TGF-beta is a potent stimulator of FN release by RPE cells, and exerts its effects via signal transduction involving PKC. Its effect is reduced by IFN-gamma.

The presence of constituents of the renin-angiotensin system (RAS) in ocular tissues and fluids suggests this system is involved in ocular physiology. Angiotensin II (AngII) is the main biological effector of the system, so we measured AngII in plasma and in aqueous humor of the anterior ocular chamber of patients undergoing cataract extraction. Untreated normotensive patients were compared with arterial hypertensive patients taking either diuretics which stimulate the RAS or angiotensin converting enzyme (ACE) inhibitors which reduce the production of AngII. Plasma levels of AngII were higher in patients on diuretics (5.46 +/- 1.04 fmol/ml; mean +/- SEM) than in untreated cataract patients (2.28 +/- 0.32 fmol/ml, p < 0.02), and were very low with ACE inhibitors (0.51 +/- 0.18 fmol/ml). In aqueous humor, AngII was measurable in 7 of 11 patients on diuretics (median 1.1 fmol/ml), and in 6 of 16 normotensive patients (median < 0.55 fmol/ml), but not in aqueous humor of 4 patients receiving enalapril or captopril. These results demonstrate the presence of AngII in the eye but do not exclude either its sequestration in the eye or local production. The possibility of individual measurements of intraocular AngII will permit more precise determination of its role in future studies.

To determine whether immunoglobulins of the IgG class diffus up to the corneal center after subconjunctival injection, rabbits were injected with fluorescein-isothiocyanate-labeled human IgG. The inoculum diffused from the entire periphery centrepetally towards the corneal center. The progression of the diffusion front slowed down as the distance to the limbus increased. The first increase of fluorescence in the corneal center was observed on day 6. The intensity increased during the following 10 days despite resorption in the corneal periphery due to the flow of IgG from paracentral toward central areas. The diffusion coefficient of 0.003-0.004 cm2/day was calculated by computer simulation using Fickian diffusion equations adapted for corneal geometry. We conclude that after subconjunctival application, IgG diffuses up to the corneal center with a delay of several days and that the penetration speed decreases as the distance to the limbus increases. This kinetics contributes to our understanding of the role of IgG in corneal pathology and may help to design therapeutic schedules for immunotherapy with IgG.

These observations underline the importance of protein leakage through a damaged blood-ocular barrier and of direct contact of monocytes/macrophages with RPE cells, as well as their reciprocal potentiating effect on RPE cell proliferation. Thus, early stabilization of the blood-ocular barrier, which would preclude or reduce protein leakage and invasion of inflammatory cells into the eye, could be a target for pharmacologic prevention of PVR.

No abstract