人CD4+CD25+调节性T细胞系的建立与功能分析

目的：建立可在体外长期培养的人CD4^ CD25^ 调节性T细胞系并研究其免疫生物学特性。方法：用流式分选的方法从健康人外周血淋巴细胞中得到CD4^ CD25^ T细胞，并对其进行体外长期培养、扩增；淋巴细胞转化实验分析其免疫抑制功能，流式细胞法分析其表型。结果：经对人CD4^ CD25^ T细胞的长期培养扩增，获得具有免疫抑制功能的调节性T细胞系。该T细胞系对经TCR的刺激不敏感，且能抑制同一来源或同种异型CD4^ CD25^ T细胞的活化，大剂量IL－2可以逆转其抑制功能。长期培养的CD4^ CD25^ T细胞，膜表面CD25和CTLA－4分子持续高表达，而CD4^ CD25^ T细胞CD25和CTLA－4分子的表达呈周期性变化。结论：将CD4^ CD25^ T细胞体外扩增培养成系，其功能和表型与同一来源的CD4^ CD25^ T细胞显著不同。

In vitro generation and function analysis of CD4 + CD25+ regulatory T cell from the healthy volunteers

Objective:To establish and characterize human CD4 +CD25 + T cell lines Methods:CD4 +CD25 + T cells were isolated from healthy volunteers and expanded in vitro Their suppressive effect on the proliferation of CD4 + T cells was analyzed in proliferation assays Phenotype of the T cell lines was determined by FACS Results:Long term cultured CD4 +CD25 +T cells were hyporesponsive to stimulation via their T cell receptor In co culture experiments CD4 +CD25 + T cells could suppress the activation and proliferation of CD4 + T cells, and the suppression was abrogated when exogenous IL 2 was provided In vitro expanded human CD4 +CD25 + T cells remained positive for CD25 and CTLA 4 expression Conclusion:CD4 +CD25 + T cells can be established as long term cultured T cell lines while still maintaining their suppressive function and distinctive phenotype from CD4 +CD25 - T cells