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目的制备重组耻垢分枝杆菌(recombinant Mycobacterium smegmatis,rM.smegmatis)NY-ESO-1疫苗,并分析其在小鼠体内的免疫原性。方法从人睾丸m RNA中扩增NY-ESO-1基因,克隆至携带β半乳糖苷酶信号肽的表达载体,转化M.smegmatis,构建r M.smegmatis。以rM.smegmatis经腹腔免疫小鼠,200μl/只,于第12周用NY-ESO-1加强免疫1次,第3、6、9、12、13周经尾静脉采血1次,分离血清,并于第13周取小鼠脾脏,制备脾淋巴细胞,检测小鼠血清中NY-ESO-1抗体及其亚型、CD4~+、CD8~+T细胞比例、脾细胞增殖能力、IFNγ及IL-2的分泌水平及体外CTL杀伤能力。结果加强免疫后,与NY-ESO-1组比较,rM.smegmatis-p LA71NY组小鼠血清抗体水平差异无统计学意义(P>0.05),CD4请下载后查看，本文暂不支持在线获取查看简介。+及CD8~+T细胞比例、IFNγ及IL-2的分泌水平及体外CTL杀伤能力均显著升高(P<0.05),脾细胞的增殖能力显著下降(P<0.05)。结论 rM.smegmatis NY-ESO-1疫苗在小鼠体内具有良好的免疫原性,为该疫苗用于肿瘤的免疫治疗奠定了基础。
Objective To prepare a recombinant Mycobacterium smegmatis (NY) sMgmatis NY-ESO-1 vaccine and analyze its immunogenicity in mice. Methods NY-ESO-1 gene was amplified from human testis m RNA, cloned into the expression vector carrying β-galactosidase signal peptide, and transformed into M.smegmatis to construct r M.smegmatis. The mice were immunized intraperitoneally with rM.smegmatis at 200μl / mouse, once at week 12 with NY-ESO-1, and at the third, sixth, ninth, twelfth and thirteenth weeks, Spleen lymphocytes were prepared from the spleen of the mice on the 13th week. The levels of NY-ESO-1 antibody and its subtypes, the ratio of CD4 ~ + and CD8 ~ + T cells, the splenocyte proliferation ability, IFNγ and IL -2 secretion and CTL cytotoxicity in vitro. Results Compared with NY-ESO-1 group, there was no significant difference in serum antibody levels between rM.smegmatis-p LA71NY group and control group after immunization (P> 0.05). CD4 download this article for review, this article does not support online access to view Introduction. (P <0.05). The proliferation of spleen cells was significantly decreased (P <0.05). Conclusion The rM.smegmatis NY-ESO-1 vaccine has good immunogenicity in mice and lays a foundation for immunotherapy of the vaccine.