关键词: 分枝杆菌，结核;疫苗，DNA;基因;免疫性;免疫原性;Ag85B

    摘 要:目的 研究已构建的编码结核分枝杆菌(Mycobac-terium tuberculosis，MTB)H37Ra株Ag85B成熟蛋白基因的真核表达质粒pTB30m的表达和免疫原性. 方法 以电穿孔法将pTB30m转染CHO细胞，RT-PCR法检测转染细胞中Ag85B特异的mRNA的表达.将pTB30m质粒肌注免疫BALB/c和C57BL/6小鼠，ELISA法检测基因免疫的体液免疫应答效果.分离免疫小鼠的脾淋巴细胞，体外用抗原再刺激，生物学方法检测IFNγ 产生水平和MTT法检测淋巴细胞增殖. 结果 RT-PCR检测证实pTB30m可在CHO细胞中表达.pTB30m免疫小鼠可引起较高水平的Ag85B特异性的抗体应答.免疫小鼠的脾淋巴细胞，体外用抗原再刺激，淋巴细胞增殖显著，BALB/c和C57BL/6小鼠IFNγ 的量分别达到1360ng L-1 和1965ng L-1 ，而生理盐水和空质粒对照组均低于80ng L-1 . 结论 应进一步研究pTB30m作为TB的基因疫苗用于TB的防治的效果.

    Gene vaccination of Mycobacterium tuberculosis DNA vaccine encoding mature form of Ag85B

    FAN Xiong-Lin，XU Zhi-Kai，LI Yuan，XUE Ying，ZHANG Fang-Lin，LI Bie-Hu，BAI Guang-Chun

    Department of Microbiology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China

    Keywords:Mycobacterium tuberculosis;vaccines，DNA;genes;immunity;immunogenicity;Ag85B

    Abstract:AIM To investigate the expression and immuno-genicity of recombinant eukaryotic expression plasmid pTB30m encoding mature form of Ag85B of Mycobacterium tuberculosis H37Ra strain.METHODS pTB30m was trans-fected into CHO cells by electroporation ......