重组耻垢分枝杆菌NY-ESO-1疫苗的制备及其免疫原性分析  

Preparation and of immunogenicity of recombinant Mycobacterium smegmatis NY-ESO-1 vaccine

作　　者：陈丹丹[1] 高丹[1] 瞿爱东[1] 徐帆洪[1] 

机构地区：[1]上海生物制品研究所有限责任公司,上海200052

出　　处：《中国生物制品学杂志》2015年第12期1233-1237,1244,共6页Chinese Journal of Biologicals

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(P>0.05),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(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 recombinant Mycobacterium smegmatis NY-ESO-1 vaccine and analyze its immunogenicity in mice. Methods NY-ESO-1 gene was amplified from human testis m RNA and cloned into the Mycobacterium expression vector containing β-lactamase signal peptide, and the constructed recombinant plasmid was transformed to M. smegmatis. Mice were immunized i. p. with the obtained recombinant M. smegmatis（r M. smegmatis）, 200 μl for each, and boosted 12 weeks later with NY-ESO-1. Venous blood samples were collected at weeks 3, 6, 9, 12 and 13 respectively, from which sera were separated. The spleens of mice were collected at week 13, from which lymphocytes were prepared. The NY-ESO-1 antibody and its subtype in sera, the ratio of CD4~＋and CD8~＋T cells, the proliferation ability of splenic lymphocytes, the secretion levels of IFNγ and IL-2 as well as the killing ability of CTL in vitro were determined. Results As compared with those by NY-ESO-1, the serum antibody level of mice after booster immunization by r M. smegmatis-p LA71 NY showed no significant difference（P〈0. 05）, while the ratio of CD4 ＋ to CD8 ＋ T cells, the secretion levels of IFNγ and IL-2 and killing ability of CTL in vitro increased significantly（P〈0. 05）, and the proliferation ability of splenic lymphocytes decreased significantly（P〈0. 05）. Conclusion The r M. smegmatis showed good immunogenicity in mice, which laid a foundation of application of the vaccine in immune therapy of tumors.

关 键 词：重组耻垢分枝杆菌 NY-ESO-1 免疫原性 

分 类 号：R378.91[医药卫生—病原生物学] R392-33[医药卫生—基础医学]