模式抗原Mannan-TTC的制备及免疫原性分析

发布时间：2018-04-30 07:26

本文选题：甘露聚糖 + 免疫原性　；参考：《中国免疫学杂志》2013年03期

【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。
[Abstract]:Aim: to prepare a model antigen Mannan-TTC with high immunogenicity and analyze its immunogenicity. Methods: Mannan was chemically crosslinked with tetanus toxin C (TTC) protein, and the crosslinked products were analyzed by SDS-PAGE. The ability of DC loaded Mannan-TTC to stimulate the proliferation of allogeneic T cells and the ability of TTC specific CD4 T cells to proliferate and secrete IFN- 纬 were detected by mixed lymphocyte reaction and ELISA was used to detect the level of TTC specific antibody. Results: the model antigen Mannan-TTCwas obtained by successfully crosslinking Mannan with TTC. By activating dendritic cells, the crosslinked product not only enhanced the proliferation of allogeneic T cells, but also significantly enhanced the proliferation and secretion of IFN- 纬, a cytokine of CD4 T lymphocytes specific to TTC antigen. Animal immunoassay showed that the cross-linked product could stimulate the immune cells of mice to produce TTC antigen-specific antibodies, but there was no significant difference compared with the stimulation of TTC antigen alone. Conclusion: the obtained Mannan-TTC antigen has good immunogenicity and can significantly enhance the cellular immune response, which provides a good model antigen for the study of antigen presentation and immune response in vivo and in vitro.
【作者单位】：新乡医学院医学检验系;新乡医学院第一附属医院检验科;新乡医学院第三附属医院检验科;
【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
【分类号】：R392.1

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