重组耻垢分枝杆菌NY-ESO-1疫苗的制备及其免疫原性分析

发布时间：2018-04-23 08:00

本文选题：重组耻垢分枝杆菌 + NY-ESO-　；参考：《中国生物制品学杂志》2015年12期

【摘要】：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(P0.05),CD4~~+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(P0.05),脾细胞的增殖能力显著下降(P0.05)。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。
[Abstract]:Objective to prepare the recombinant recombinant Mycobacterium smegmatis (rM.smegmatis) NY-ESO-1 vaccine and analyze its immunogenicity in mice. Methods the NY-ESO-1 gene was amplified from m RNA in human testis and was cloned to carry the expression body of beta galactosidase signal peptide and transform M.smegmatis, and R M.smegmatis. was constructed with rM.smegmat. Is was immunized in the abdominal cavity of mice, 200 mu l/ only, the immunization was strengthened by NY-ESO-1 for 1 times in twelfth weeks, 1 times by the tail vein, and the spleen was isolated from the tail vein for 1 weeks. The spleen lymphocyte was prepared for thirteenth weeks, and the NY-ESO-1 antibody and its subtype in the serum of mice, the proportion of CD4~+, CD8~ +T cells, the splenocytes proliferation, IFN gamma and IL-2 secreting water were detected. Compared with the NY-ESO-1 group, there was no significant difference in the serum antibody level of the rM.smegmatis-p LA71NY group (P0.05), the proportion of CD4~~+ and CD8~+T cells, the level of IFN gamma and IL-2 and the killing ability of CTL in rM.smegmatis-p LA71NY group (P0.05), and the proliferation ability of spleen cells decreased significantly (P0.05). Conclusion rM.smegmatis NY-ESO-1 vaccine has good immunogenicity in mice and lays a foundation for the immunotherapy of tumor.

【作者单位】：上海生物制品研究所有限责任公司;
【分类号】：R378.91;R392

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