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目的:观察常用丝裂原ConA对CD4+CD25+调节性T细胞抑制功能的影响与早期活化标志CD69之间的关系。方法:用免疫磁珠分离BALB/c小鼠脾脏CD4+CD25+调节性T细胞、CD4+CD25-效应性T细胞,加入不同浓度的ConA,12 h后收集细胞,流式细胞术分析其CD69的表达。CFSE标记CD4+CD25-效应性T细胞,按2∶1比例与CD4+CD25+调节性T细胞共同培养,培养同时加ConA,3 d后流式细胞仪分析ConA对CD4+CD25+调节性T细胞抑制CD4+CD25-效应性T细胞增殖的影响。结果:ConA能剂量依赖性地升高CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞CD69的表达,对两群细胞CD69的升高表现出了相似的作用;另外,ConA在上述浓度能剂量依赖激活CD4+CD25+调节性T细胞的抑制功能。结论:ConA在体外对CD4+CD25+调节性T细胞活化和功能具有促进作用。ConA能一定程度模拟抗原,因而可用于评价药物对调节性T细胞的活化和功能影响。
Objective: To observe the effect of common mitogen ConA on CD4 (superscript +) CD25 (superscript +) regulatory T cell suppressive function and its relationship with early activation marker CD69. Methods: The splenic CD4 + CD25 + regulatory T cells and CD4 + CD25- effector T cells of BALB / c mice were isolated by immunomagnetic beads. ConA was added into the spleen of BALB / c mice at different concentrations for 12 h. The cells were collected after 12 h. expression. CFSE-labeled CD4 + CD25- effector T cells were co-cultured with CD4 + CD25 + regulatory T cells in a 2: 1 ratio and ConA supplemented with ConA for 3 days. Flow cytometry was used to analyze the inhibition of CD4 + CD25 + regulatory T cells by flow cytometry CD4 + CD25-effector T cell proliferation. Results: ConA could increase the expression of CD69 on CD4 + CD25 + regulatory T cells and CD4 + CD25- effector T cells in a dose-dependent manner, which showed a similar effect on the increase of CD69 in both groups. In addition, Concentration can dose-dependently activate the suppressive function of CD4 + CD25 + regulatory T cells. CONCLUSION: ConA can promote the activation and function of CD4 + CD25 + regulatory T cells in vitro. ConA mimics antigens to some extent and thus can be used to assess the effects of drugs on the activation and function of regulatory T cells.