Abstract
We previously reported that Ia+ monolayers of LEW rat corneal endothelial (CE) cells were unable to stimulate proliferation of MHC compatible T cell lines or IL-2 release from hybridomas, and inhibited [3H]-thymidine incorporation when added to conventional lymphocyte proliferation assays. Our purpose was to further analyze the mechanism of the inhibitory activity of CE cells on T lymphocyte activation. Mitogen-induced proliferative responses of splenocytes were found to be as susceptible to inhibition by CE cells as previously reported for antigen-specific activation of T cell lines. Antigen presenting cell (APC) antigen pulsing experiments showed that CE cells did not inhibit antigen processing. Flow cytometry and microscopic observation of the co-cultures revealed that T cells became activated in the presence of antigen, APC and CE cells, exhibiting morphologic changes of blast cell formation, although they did not divide unless given exogenous IL-2. However, if T cells were preactivated in the absence of CE cells, they were no longer susceptible to inhibition if subsequently transferred into CE cell-conditioned medium or onto CE cells. Evidence for an inhibitory factor in CE cell culture supernatant was revealed by two approaches: 1) addition of conditioned medium from CE cell cultures led to inhibition of lymphocyte proliferation assays, and 2) split-well assays also demonstrated the existence of a cell-free immunosuppressive factor produced by the CE cells. However, the inhibition mediated by supernatant alone was less potent than that by direct T cell contact with CE cells, implying that cell-cell interaction contributed to the inhibition. Indomethacin, a prostaglandin synthetase inhibitor, did not reverse CE cell-mediated inhibition. Neutralizing antibodies to TGF-β1 and 2 did not reverse the inhibition by CE cells. In summary, T cells received activation signals from APC in the presence of CE cells, but proliferation was inhibited unless exogenous lymphokine was added.
Original language | English (US) |
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Pages (from-to) | 575-585 |
Number of pages | 11 |
Journal | Current Eye Research |
Volume | 13 |
Issue number | 8 |
DOIs | |
State | Published - 1994 |
Bibliographical note
Funding Information:The authors thank Dr John W. Torseth and Shiv A. Prasad for their critical review of the manuscript. This work was supported by NIH grants EY05417 and 09207 (D.S.G.), Research to Prevent Blindness, Inc., the Minnesota Lions and Lionesses Clubs, the Yoshida Science Technology Foundation (H.K.) and the Nippon Eye Bank Association (H.K.). D.S.G. is a Research to Prevent Blindness Senior Scientific Investigator.
Keywords
- Cell culture
- Corneal endothelial cells
- Experimental autoimmune uveoretinitis
- Immune privilege
- Local immunoregulation
- Rat