含噻唑烷-4-酮的免疫刺激剂CH2a增强人iNKT细胞的功能  

Immunostimulator CH2a bearing thiazolidin-4-one promotes the function of human iNKT cells in vitro

作　　者：孟明[1,2] 李春晓[1] 王泽瑞[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,河北保定071000 [2]河北大学化学与环境科学学院,化学生物学河北省重点实验室,河北保定071000

出　　处：《细胞与分子免疫学杂志》2013年第4期345-349,共5页Chinese Journal of Cellular and Molecular 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);河北大学大学生科技创新项目(201210075005)

摘　　要：目的研究含噻唑烷-4-酮的糖类衍生小分子免疫刺激剂CH2a对人iNKT细胞功能的影响。方法分离健康人外周血单个核细胞(PBMC),经α-半乳糖神经酰胺(α-Galcer)和IL-2体外扩增后,利用磁珠分选分离纯化得到恒定型自然杀伤T细胞(iNKT细胞)。5(6)-羧基二乙酸荧光黄琥珀酰亚胺酯(CFDA-SE)标记的纯化iNKT细胞,与CH2a共孵育后,流式细胞术检测细胞增殖和分代,IFN-γ和IL-4的表达;MTT法定量检测细胞增殖率及对K562细胞的杀伤活性。ELISA检测CH2a处理细胞培养基中IFN-γ和IL-4水平。结果 CH2a能显著促进IL-2引起的iNKT细胞体外增殖;提高IFN-γ和IL-4的生成量及IFN-γ/IL-4的比值;并显著增强iNKT细胞的杀伤活性。结论 CH2a有可能通过诱导iNKT分泌Th1类的细胞因子,调节T细胞向Th1分化,从而增强细胞免疫功能;同时显著提高iNKT的细胞毒功能。Objective To explore the immunostimulatory effect of immunostimulant CH2a bearing thiazolidin-4-one on human invariant natural killer T （iNKT） cell function in vitro. Methods Peripheral blood mononuclear cells （PBMCs） were obtained from healthy adults, and then amplified with α-Galcer and IL-2 in vitro. The iNKT cells were isolated from these pro- liferating cells by magnetic-activated cell sorting （MACS） method. The purified iNKT cells were labled with 5-（ and 6）-carboxyfluorescein diacetate succinimidyl ester （CFDA-SE） and then incubated with CH2a for functional analysis, including cell proliferation, expressions of IFN-）, and IL-4 by flow cytometry, proliferation rate and cytotoxicity by MTT assay. In additon, ELISA was performed to determine the levels of IFN-γ and IL-4 in cell culture media. Results CH2a significantly promoted the proliferation of iNKT cells induced by IL-2 in vitro, stimulated the release of both IFN-r and IL-4, and led to the increase in IFN-γ/IL-4 ratio. More importantly, the cytotoxicity of iNKT was also markedly elevated under the stimulation of CH2a. Conclusion Immunostimulant CH2a probably promote the production of Th1-like cytokines and the differentiation of Th0 to Th1, so as to improve cellular immune function. In addition, CH2a could significantly enhance the cytotoxicity of iNKT cells.

关 键 词：INKT细胞 磁珠分选 IFN-Γ IL-4 免疫刺激剂 

分 类 号：R392.12[医药卫生—免疫学] R256[医药卫生—基础医学]