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Hiroshi Keino, Masaru Takeuchi, Takeshi Kezuka, Takaaki Hattori, Masahiko Usui, Osamu Taguchi, J. Wayne Streilein, Joan Stein-Streilein; Induction of Eye-Derived Tolerance Does Not Depend on Naturally Occurring CD4+CD25+ T Regulatory Cells. Invest. Ophthalmol. Vis. Sci. 2006;47(3):1047-1055. doi: 10.1167/iovs.05-0110.
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purpose. Regulatory CD4+ T cells (T regs) arise in the spleens of mice with anterior chamber–associated immune deviation (ACAID), an eye-derived tolerance evoked by injection of antigen into the ocular anterior chamber (AC). The current study was conducted to investigate the possibility that these T regs express CD25 and are derived from natural CD4+CD25+ T cells.
methods. Naïve T cells from DO11.10 mice were activated in vitro by ovalbumin (OVA)-pulsed, TGFβ-treated antigen-presenting cells (APCs), and the expression of CD25 assayed by flow cytometry. OVA-specific ACAID T regs were obtained from the spleens of DO11.10 mice with ACAID to OVA. Immunomagnetic enrichment was used to sort out CD4+CD25+, and CD4+CD25− ACAID T cells before they were injected into OVA-immunized mice or examined for mRNA expression of the regulatory T-cell transcription factor Foxp3. In addition, before AC injection of OVA, systemic depletion of CD25+ T cells was performed with injections of anti-IL-2 receptor antibody into the mice.
results. OVA-specific T cells from DO11.10 mice expressed CD25 when exposed to OVA-pulsed, TGFβ-treated APCs, even when the DO11.10 T cells were depleted of CD25+ cells before their in vitro stimulation. In addition, DH was suppressed in naïve mice that were injected with CD4+CD25+ or CD4+CD25− ACAID T cells. The CD4+CD25+, but not the CD4+CD25−, ACAID T regs expressed Foxp3. Finally, OVA induced ACAID in mice depleted of CD25+ cells.
conclusions. Some of the CD4+ T regs of ACAID arise from CD25− precursors, and the induction of ACAID is not dependent on the presence of natural CD4+CD25+ T regs.
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