Background Regulatory CD4+CD25+ T cells have been proven to be essential for maintenance of peripheral tolerance and autoimmune diseases. ACAID is a model of immune privilege in the eye. Relatively little is known about the role and phenotype of these regulatory CD4+CD25+ T cells in ACAID. Methods Injection of OVA into the anterior chamber of BALB/C mice was performed to induce ACAID. The frequencies of splenic CD4+CD25+ Tregs and the expression of CTLA-4 and LAG-3 on these cells were determined by flow cytometry. Magnetic cell sorting was used to isolate CD4+CD25+ and CD4+CD25¿T cells. The function of CD4+CD25+ T cells was detected by in vitro immunosuppression assays and in vivo adoptive transfer. Results ACAID was successfully induced following an i.c. injection of OVA. Frequencies of CD4+CD25+ and Tregs were significantly increased in ACAID mice as compared to those in controls. The CD4+CD25+ T cells stimulated with OVA in ACAID mice showed a stronger suppressive ability in vitro than those seen in non-ACAID mice. CD4+CD25+ T cells from ACAID mice, but not from non-ACAID mice, were able to suppress DTH responses in an antigen-specific manner following adoptive transfer. The frequencies of CTLA-4 or LAG-3 on Tregs in ACAID mice were higher as compared with those in naive mice. Conclusion Splenic CD4+CD25+Foxp3+T cells expressing CTLA4 and LAG3 play an important role in the induction of ACAID
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