模式抗原Mannan-TTC的制备及免疫原性分析  

Preparation of mode antigen Mannan-TTC and analysis of its immunogenicity

作　　者：王凡平[1] 王明永[1] 赵东方[2] 马淑君[1] 余文发[1] 杜继英[1] 丁肖华[1] 朱琳琳[1] 谭静[3] 赵永新[3] 毛光兰[3] 申家辉[3] 

机构地区：[1]新乡医学院医学检验系,新乡453003 [2]新乡医学院第一附属医院检验科,新乡453002 [3]新乡医学院第三附属医院检验科,新乡453003

出　　处：《中国免疫学杂志》2013年第3期295-299,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(31100645);河南省科技攻关计划项目(102102310154);河南省高校科技创新人才项目(2012HASTIT024);河南省高校青年教师骨干项目(2010GGJS-117);河南省教育厅科学技术研究重点项目(12A320017)资助项目;新乡医学院重点学科开放基金(ZD200928)资助项目

摘　　要：目的:制备高免疫原性的模式抗原Mannan-TTC,分析其免疫原性。方法:将甘露聚糖(Mannan)与破伤风毒素C片段(TTC)蛋白进行化学交联,SDS-PAGE分析交联产物。用混合淋巴细胞反应检测DC负载Mannan-TTC刺激同种异体T细胞增殖的能力;WST-1和ELISA检测TTC特异性CD4+T细胞的增殖和分泌IFN-γ的能力;小鼠免疫后,用ELISA检测TTC特异性抗体水平。结果:成功交联Mannan与TTC,获得模式抗原Mannan-TTC。该交联产物能通过活化树突状细胞(DC),不仅增强了对同种异体T细胞增殖的能力,而且显著增强TTC抗原特异性的CD4+T淋巴细胞增殖及分泌细胞因子IFN-γ。动物免疫实验则显示,该交联产物能刺激小鼠免疫细胞产生TTC抗原特异性的抗体,但与单纯TTC抗原刺激相比,无显著性差异。结论:所获Mannan-TTC抗原免疫原性好,能显著增强细胞免疫应答,为研究体内外抗原提呈、免疫应答提供了良好的模式抗原。Objective：To preparation of mode antigen Mannan-tetanus toxin fragment C （TTC） and analysis of its immunoge- nicity. Methods：Mannan was conjugated to TTC, thus Mannan-TrC antigen was obtained. The cross-linked product was identificated by SDS-PAGE. The ability to stimulate the proliferation of allogenie T cells by Mannan-TYC-stimulated DCs was detected by mixed lym- phocyte reaction（MLR）. WST-1 and ELISA were used to analyse the proliferation and IFN-＇y secretion in TTC antigen-specific CD4 ＋ T lymphocyte. ELISA was used to detect the levels of TTC-speeific antibodies by using immunized mice. Results： Mannan was conjuga- ted to TTC successfully, and mode antigen Mannan-TTC was obtained. Through activation of dendritic cells （ DC）, Mannan-TTC could not only increasingly stimulate allogenetic T cells proliferation, but also enhancively stimulate TTC antigen-specific CD4 ＋ T lymphocyte proliferation and secretion of IFN-γγ The mice immunization experiments revealed that the Mannan-TTC antigen could stimulate the mousse immune cell produce TTC antigen-specific antibody, but there was no significant difference compared with the control group （TYC group）. Conclusion：Mode antigen Mannan-TTC was obtained successfully, which can significantly enhance cellular immune re- sponses, and provide a good model antigen for studying antigen presentation and immune responses.

关 键 词：甘露聚糖 免疫原性 模式抗原 

分 类 号：R392-33[医药卫生—免疫学]