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Min Sun, Peizeng Yang, Liping Du, Hongyan Zhou, Xiangrong Ren, Aize Kijlstra; Contribution of CD4+CD25+ T Cells to the Regression Phase of Experimental Autoimmune Uveoretinitis. Invest. Ophthalmol. Vis. Sci. 2010;51(1):383-389. doi: 10.1167/iovs.09-3514.
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To investigate the role of CD4+CD25+ Treg cells in the development of experimental autoimmune uveoretinitis (EAU).
EAU was induced in B10RIII mice by immunization with IRBP161–180 in complete Freund's adjuvant and evaluated clinically and pathologically on days 0, 7, 14, 21, and 28. Lymphocytes from draining lymph nodes (LNs) were subjected to flow cytometry to analyze the frequency of CD4+CD25+ Treg cells. CD4+CD25+ Treg cells and CD4+CD25− T cells were separated by means of magnetic-assisted cell sorting and cocultured or crossover cultured for 3 days. Proliferation of CD4+CD25− T cells was measured using a modified MTT assay. The levels of IFN-γ and IL-17 in the supernatants were determined by enzyme-linked immunosorbent assay.
Clinical and histopathologic results showed a severe intraocular inflammation in the immunized mice. The frequency of CD4+Foxp3+ T cells and CD4+CD25+Foxp3+ T cells in the draining LN lymphocytes was increased on day 7, reached its peak on day 14, and maintained a high level up to day 42. CD4+CD25+ Treg cells obtained from mice on days 14 and 28 after immunization showed a stronger inhibitory effect on the proliferation of CD4+CD25− T cells and the production of IFN-γ by CD4+CD25− T cells compared with those obtained from control mice. CD4+CD25+ Treg cells did not affect IL-17 production. Transfer of CD4+CD25+ Treg cells obtained from EAU mice was able to suppress EAU induction by IRBP161–180 that was not observed after transfer of cells from mice that had received CFA alone, suggesting antigen specificity of the Treg response.
A significantly increased frequency and immunoregulatory action of CD4+CD25+ Treg cells is associated with the development and regression of EAU, suggesting that CD4+CD25+ Treg cells are induced during EAU and may be involved in its regression.
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