猪CD4+CD25+treg细胞系的建立与功能分析

目的 分析猪淋巴细胞表型,分离猪CD4 CD25 T调节性T细胞系并鉴定其免疫生物学特性.方法用磁珠双阳性分选的方法从健康猪外周血及脾脏淋巴细胞中得到CD4 CD25 T细胞CD4 CD25-T细胞,监测其foxp3的表达,并对其进行体外长期培养、扩增,混合淋巴细胞培养实验分析其免疫抑制功能,流式细胞法分析其表型变化.结果猪CD4 CD25 T细胞与人类和啮齿类动物一样,是具有免疫抑制功能的调节性T细胞系.该T细胞系foxp3基因同样高表达,且能抑制同基因CD4 CD25-T细胞的活化,大剂量IL-2可以逆转其抑制功能,扩增培养的猪CD4 CD25 T细胞和诱导扩增的CD4 CD25-T细胞均具有免疫抑制功能.结论 猪CD4 CD25 T细胞为调节性T细胞,具有免疫抑制功能.

In vitro generation and function analysis of porcine CD4 + CD25 + T-regulatory cell

Objective To analyze the proportion of porcine peripheral blood lymphocyte, establish and characterize porcine CD4 CD25 T cell lines. Methods CD4 CD25 and CD4 CD25-T cells were isolated by multi sorts beads method from healthy porcines and expanded in vitro. The foxp3 expression was monitored by RT-PCR and its suppressive effects to the proliferation of CD4 CD25-T cells were analyzed by proliferation assay. Phenotype of the T cell lines was determined by FACS. Results The porcine fresh CD4 CD25 T cells were the same as these of human and rodents. In co-culture experiments, CD4 CD25 T cells having a high foxp3 expression could suppress the activation and proliferation of CD4 CD25-T cells, which would abrogated when exogenous IL-2 was provided. Both in vitro expanded CD4 CD25- and CD4 CD25- T cells possessed the suppressive function. Conclusion The porcine CD4 CD25 T cells are T-regulation cells and possess immune inhibiting function.