The interaction between organ-resident cells from the anterior uvea of the eye and T helper (Th) cells was investigated. Cells from Lewis rat ciliary body processes (CB cells), grown in tissue culture using an explant technique, could be induced to express major histocompatibility complex class II (Ia) antigens by incubation with rat interferon-gamma. Ia+ CB cells only poorly functioned as antigen-presenting cells (APC) for a syngeneic, uveitogenic Th cell line specific for the retinal soluble antigen (SAg). Moreover, if added to an Ag-driven lymphocyte proliferation assay in the presence of conventional APC, the rat CB cells had an inhibiting effect on Th proliferation. This inhibitory activity was not species specific, since similar effects were observed with bovine and human ciliary epithelial cells. The suppressive activity of CB cells was composed of a soluble factor, as well as a membrane-associated inhibitor. The soluble activity did not appear to be related to transforming growth factor-beta (TGF-beta), since no reversal of inhibition by a neutralizing antibody to TGF-beta was found. Part of the soluble inhibitory activity could be reversed by indomethacin treatment. The membrane-associated component was trypsin sensitive, suggesting a protein molecule. After abrogation of the inhibitory capacity by trypsin treatment and fixation by glutaraldehyde, CB cells effectively presented SAg to Th cells. These data suggest that CB cells are capable of mediating both Ag presentation and inhibition of Th cell proliferation.
Systemic injection of bacterial endotoxin (Lipopolysaccharide, LPS) in experimental animals induces anterior uveitis without major pathological changes in other organs. The present study investigates the effect of LPS on production of inflammatory mediators in cultured bovine pigmented ciliary epithelial cells (CB-cells) by means of radioimmunoassays and bioassays. LPS was found to stimulate CB-cells to secrete prostaglandin E2 and prostacyclin (assayed as its stable metabolite 6-keto-prostaglandin F1a), but not leukotriene B4 or thromboxane A2 (assayed as its stable metabolite thromboxane B2). CB-cells produced membrane-associated interleukin 1-activity in response to LPS, but no tumor necrosis factor-activity was found after challenge of CB-cells with LPS. The direct effect of LPS on production of inflammatory mediators by cells from the anterior uvea could play a role in the pathophysiology of endotoxin-induced uveitis.
We compared the abilities of young and senescent fibroblasts to take up and degrade [3H]ribonuclease A (native and oxidized), [3H]ribonuclease4-13, [3H]hemoglobin, [3H]glyceraldehyde-3-phosphate dehydrogenase, [3H]beta-galactosidase, [3H]glycogen phosphorylase, and [125I]serum albumin. The endocytic uptake of these proteins ranged from fluid-phase to predominantly absorptive. Intralysosomal degradation rates of the different endocytosed proteins varied by an order of magnitude, but in no case was there a difference between cultures of young and senescent fibroblasts.
The expression of major histocompatibility complex (MHC) class II antigens in cultured bovine ciliary epithelial cells was investigated by means of indirect immunohistochemistry and immunocytofluorometry. Ciliary epithelial cells grown in control tissue-culture medium did not express MHC class II. However, after incubation with bovine gamma-interferon (IFN-G) in concentrations as low as 0.3 units/ml, nearly all cells stained for MHC class II. Tumor necrosis factor increased IFN-G-induced MHC class II expression. A reduction in IFN-G-induced MHC class II expression was observed with dexamethasone, prostaglandin E2 and alpha-interferon. To test whether MHC class II expression in response to IFN-G was specific for the ciliary epithelium, several intraocular tissues were grown in culture and incubated with IFN-G. MHC class II expression was observed in all tissues tested for response to IFN-G, but at different sensitivities. Retinal pigment epithelium and ciliary epithelium exhibited the highest sensitivity, followed by corneal endothelium and lens epithelium; the lowest sensitivity was observed for retinal vascular pericytes. The results are discussed in the context of MHC class II expression on the ciliary epithelium in anterior uveitis.
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