免疫调节剂CH2b对活动期类风湿关节炎患者iNKT细胞免疫调节功能的影响  

Effects of a novel synthetic immunostimulator CH2b on iNKT cells isolated from patients with active rheumatoid arthritis

作　　者：孟明[1] 杨飞[1] 许鸣华[2] 陈丹[1] 候明辉[2] 刘嘉琳[1] 陈冬志[1] 

机构地区：[1]河北大学医学部,保定071000 [2]河北大学附属医院

出　　处：《中华微生物学和免疫学杂志》2015年第12期916-920,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（81373197）;河北省自然科学基金（H2014201133,H2015201131）

摘　　要：目的研究新型合成的含噻唑烷4．酮的免疫调节剂CH2b对活动期类风湿关节炎（RA）患者iNKT（invariant nature killer T）细胞功能的影响。方法外周血单个核细胞（PBMC）分离自活动期RA患者，经α—Galcer和IL-2体外刺激扩增后利用iNKT分离试剂盒经磁珠分选（MACS）得到纯化的iNKT细胞；采用MTY法定量测定CH2b对iNKT细胞增殖的影响；MILLIPLEX MAP Human Cytokine／Chemokine kit检测相应iNKT细胞培养上清中IFN-γ/IL-4的水平；RT—PCR检测iNKT细胞中IFN-γ mRNA与IL-4 mRNA表达水平；Western blot检测iNKT细胞中T-bet／GATA-3表达水平。结果CH2b能显著促进IL-2活化的iNKT细胞体外增殖；增加IL-4的分泌且显著降低IFN-γ/IL-4比值；上调iNKT细胞GATA-3和IL-4 mRNA表达。结论免疫调节剂CH2b有可能通过GATA-3通路诱导iNKT细胞分泌Th2样细胞因子，促进Th0向Th2分化，提高iNKT细胞的免疫调节功能。Objective To investigate the effects of a novel synthetic immunostimulator CH2b containing thiazolidin-4-one on the function of invariant nature killer T （iNKT） cells isolated from patients with active rheumatoid arthritis （RA）. Methods Peripheral blood mononuclear cells （PBMCs） isolated from patients with active RA were in vitro cultured with α-Galcer and IL-2. The iNKT cells were separated by using magnetic activated cell sorting （MACS） method. The effects of CH2b on the proliferation of iNKT cells were analyzed by using MTT assay. MILLIPLEX MAP Human Cytokine/Chemokine kit was used to measure the levels of IFN-γ and IL-4 in the supematants of iNKT cell culture. The expressions of IFN-γand IL-4 at mRNA level in iNKT cells were analyzed by RT-PCR. Western blot assay was used to detect the levels of T-bet and GATA-3 in iNKT ceils. Results CH2b significantly enhanced the proliferation of IL-2 activated iNKT cells isolated from the patients with active RA. CH2b promoted the secretion of IL-4, resulting in a decrease in the ratio of IFN-γ/IL-4. Moreover, CH2b promoted the expressions of GATA-3 and IL-4 at mRNA level in iNKT cells. Conclusion The novel immunostimulator, CH2b, might enhance the immunoregulatory effects of iNKT cells by promoting the GATA-3 pathway-mediated secretion of Th2-1ike cytokines and inducing the differentiation of Th0 to Th2 cells.

关 键 词：类风湿关节炎 INKT细胞 T—bet GATA一3 

分 类 号：R593.22[医药卫生—内科学]