新型合成的免疫调节剂CHla促进人iNKT细胞功能的研究  

A novel synthetic immunostimulator CHla containing thiazolidin-4-one enhances the function of human iNKT cells in vitro

作　　者：孟明[1] 王泽瑞[1] 李春晓[1] 韩华[1] 史建红[1] 陈华[2] 李小六[2] 陈冬志[1] 

机构地区：[1]河北大学医学部,保定071000 [2]教育部药物化学与分子诊断省部共建重点实验室、化学生物学河北省重点实验室

出　　处：《中华微生物学和免疫学杂志》2013年第6期452-457,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（20972039）;河北省自然科学基金石药集团医药联合研究基金（B2011201169）;国家级大学生创新创业项目（201210075005）

摘　　要：目的研究新型合成的含噻唑烷4-酮环的免疫调节剂CHla对人类iNKT（invariantnaturekillerT）细胞功能的影响。方法从健康人外周血分离单个核淋巴细胞（PBMC），经α-Galeer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MTT法定量测定CHla对iNKT细胞增殖的影响，利用^51Cr释放实验观察iNKT细胞对K562细胞杀伤率的影响；利用流式细胞技术检测iNKT细胞增殖分代及细胞因子（IFN-1和IL-4）的表达。结果CHla能显著促进纯化的iNKT细胞体外增殖（以4～7代细胞数居多）；并增加IFN-γ和IL-4的释放且显著提高IFN-γ／IL-4比值；尤其是CHla能显著提高iNKT细胞的杀伤活性。结论免疫调节剂CHla有可能促进iNKT细胞分泌Th1样细胞因子，促进Th0向Th1分化。此外，CHla能够显著增强iNKT细胞细胞毒活性。Objective To investigate the immunoregulatory effects of a novel synthetic immunos- timulator CHla containing thiazolidin-4-one on the function of human iNKT（ invariant nature killer T） cells in vitro. Methods Peripheral blood mononuclear cells （PBMCs） from healthy adults were cultured with stim- ulation of c^-Galcer and IL-2 in vitro, and iNKT cells were then separated by using magnetic -activated cell sorting （MACS） method with iNKT isolation kit. The effects of CHla on the proliferation of iNKT cells were analyzed by using MTT assay, sl Cr release assay was used to evaluate the killing activity of iNKT cells to K562 cells. Fluorescence-activated cell sorting （FACS） was used to detect the proliferation of iNKT cells, as well as the production of IFN-γ/and IL-4. Results CHla was found to significantly stimulate the iNKT proliferation （ mainly up to 4-7 generations）, promote the secretion of both IFN-γ and IL-4, and increase the ratio of IFN-γ/IL-4. More importantly, CHla could markedly enhance the cytotoxieity of iNKT cells. Con- clusion Immunostimulator CHla could promote the production of Thl-like cytokines by iNKT cells and in- duce differentiation of Th0 to Thl cells. Moreover, CHla could significantly enhance the cytotoxicity of iNKT cells.

关 键 词：INKT细胞 MACS磁珠分选 IFN-Γ IL-4 免疫调节剂 

分 类 号：R9[医药卫生—药学]