Keino Hiroshi, Takeuchi Masaru, Kezuka Takeshi, Hattori Takaaki, Usui Masahiko, Taguchi Osamu, Streilein J Wayne, Stein-Streilein Joan
Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA.
Invest Ophthalmol Vis Sci. 2006 Mar;47(3):1047-55. doi: 10.1167/iovs.05-0110.
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.
Naïve T cells from DO11.10 mice were activated in vitro by ovalbumin (OVA)-pulsed, TGFbeta-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.
OVA-specific T cells from DO11.10 mice expressed CD25 when exposed to OVA-pulsed, TGFbeta-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.
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.
调节性CD4+ T细胞（Tregs）产生于具有前房相关免疫偏离（ACAID）的小鼠脾脏中，ACAID是一种通过将抗原注射到眼前房（AC）所诱发的眼部耐受性。本研究旨在探讨这些Tregs表达CD25并源自天然CD4+CD25+ T细胞的可能性。
来自DO11.10小鼠的初始T细胞在体外由卵清蛋白（OVA）脉冲处理、经转化生长因子β（TGFβ）处理的抗原呈递细胞（APC）激活，通过流式细胞术检测CD25的表达。OVA特异性ACAID Tregs从小鼠脾脏中获得，该小鼠具有针对OVA的ACAID。在将CD4+CD25+和CD4+CD25- ACAID T细胞注射到经OVA免疫的小鼠中或检测调节性T细胞转录因子Foxp3的mRNA表达之前，使用免疫磁珠富集法对其进行分选。此外，在向眼前房注射OVA之前，通过向小鼠注射抗IL-2受体抗体对CD25+ T细胞进行全身清除。
当暴露于OVA脉冲处理、经TGFβ处理的APC时，来自DO11.10小鼠的OVA特异性T细胞表达CD25，即使DO11.10 T细胞在体外刺激前已清除CD25+细胞。此外，在注射CD4+CD25+或CD4+CD25- ACAID T细胞的初始小鼠中，迟发型超敏反应（DH）受到抑制。CD4+CD25+而非CD4+CD25- ACAID Tregs表达Foxp3。最后，OVA在清除了CD25+细胞的小鼠中诱导了ACAID。
ACAID的一些CD4+ Tregs源自CD25-前体细胞，并且ACAID的诱导不依赖于天然CD4+CD25+ Tregs的存在。
