Effect of different cult ure conditions on differentiation of Treg and Th17 in PB-MC
10.3969/j.issn.1000-484X.2015.07.002
- VernacularTitle:不同培养环境对PMBC中Treg和Th17细胞分化的影响
- Author:
Hongwei SHAO
;
Hailin YANG
;
Chunmei LIANG
;
Xiaomin LU
;
Ting LIU
;
Han SHEN
;
Fenglin WU
;
Shulin HUANG
- Publication Type:Journal Article
- Keywords:
Regulatory T cells;
Th17;
Interleukin-6;
Conditioned medium
- From:
Chinese Journal of Immunology
2015;(7):874-878
- CountryChina
- Language:Chinese
-
Abstract:
Objectiv e:To investigate the effect of different culture conditions on the differentiation of Treg and Th17 to lay a foundation for exploring the methods to reverse the immune tolerance induced by tumor microenvironment.Methods:The IL-6 gene was cloned and stablely transferred into the tumor cell line expressing TGF-β.The conditioned mediums ( CM) were prepared by collecting the culture supernatants of tumor cell lines with or without IL-6 expression and used in the in vitro culture of peripheral blood mononuclear cells ( PBMC ) .The changes of Treg and Th17 in PBMC treated with different CM were detected with flow cytometry ( FCM) .Results:The expression of TGF-βin BEL-7402 was higher than that in HepG2.Thus the BEL-7402 was selected for preparation of cell line stablely transfected with IL -6 gene.ELISA detection confirmed the effective expression of IL -6 by the identified cell lines.It was showed that the Treg increased in PBMC treated with culture supernatants of tumor cells .However,the presence of IL-6 reversed the increase of Treg and promoted the differentiation of Th 17.Conclusion: The culture supernatants of tumor cells increases the proportion of Treg.However,the presence of IL-6 in this CM can reverse the increase of Treg and raise the proportion of Th 17.
