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中文摘要：
目的：研究利用免疫磁珠分选法稳定分离正常大鼠脾脏CD4^＋CD25^＋调节性T细胞的方法。方法：采用免疫磁珠两步法分离大鼠脾组织CD4^＋CD25^＋T细胞。首先采用藻红蛋白（PE）标记的抗CD25抗体和抗PE多功能磁珠试剂盒阳性分选CD25^＋T细胞,再用抗异硫氰酸荧光素（FITC）标记抗体和抗IgG磁珠阳性分选获得CD4^＋CD25^＋T细胞。分离后的细胞经流式细胞仪检测分离纯度,台盼蓝染色检测细胞存活率,体外增殖实验检测其对CD4^＋CD25^-T细胞的免疫抑制作用。结果：两次阳性分选后获得的CD4^＋CD25^＋T细胞纯度为（90.4±1.6）%,细胞存活率为（92.6±2.4）%。体外增殖实验表明,CD4^＋CD25^＋T细胞能明显抑制CD4^＋CD25^-T细胞的增殖（P〈0.01）。结论：采用免疫磁珠法两次阳性分选,可稳定地获得纯度理想并有免疫抑制功能的大鼠CD4^＋CD25^＋T细胞。
英文摘要：
Objective：To seek a stable and efficient method for isolation of splenic CD4^＋ CD25^＋ regulatory T cells （Tregs） in rats. Methods：CD4^＋ CD25^＋ Tregs were isolated from the rat splenic tissue in two steps by magnetic cell sorting （MACS） system, and CD25^＋ T cells were first positively sorted by anti-CD25 PE antibody and anti-PE multisort kit. And then CD4^＋ CD25^＋ Tregs were positively sorted by anti-CD4 FITC antibody and anti-IgG magnetic microbeads. The purity and the survival rate of sorted cells were determined by flow cytometry and trypan blue exclusion respectively, and the immunosuppressive property of CD4^＋ CD25^＋ Tregs on proliferation of CD4^＋ CD25^- T cells was also assessed by in vitro cell proliferation assay. Results：The purity of positively sorted CD4^＋ CD25^＋ Tregs was （90. 4 ± 1.6）% with the survival rate of （92.6 ± 2.4）%. The CD4^＋ CD25^＋ Tregs significantly suppressed the proliferation of CD4^＋ CD25^- T cells in mixed lymphocyte culture （P〈0. 01）. Conclusion： A twostep procedure of magnetic cell sorting system for CD4^＋ CD25^＋ Tregs isolation is established, which insures satisfactory purification of the said cells with sound function.
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