文档介绍：结核分枝杆菌Ag85B成熟蛋白基因免疫
作者:范雄林徐志凯李元薛莹张芳琳李别虎白光春
【关键词】分枝杆菌
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-,,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-,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.
Keywords:Mycobacterium tuberculosis;ines,DNA;genes;immunity;immunogenicity;Ag85B
Abstract:AIM To investigate the expression and immuno-genicity of binant eukaryotic expression plasmid pTB30m encoding mature form of Ag85B of Mycobacterium tuberculosis H37Ra pTB30m was trans-fected into CHO cells by electroporation,and expression was analyzed by RT-PCR at the level of BALB/c and C57BL/6mice were inated by pTB30m,ELISA was used to detect the antibody titer of immunized mice of IFNγ secreted by splenocytes of immunized mice up-on antigen-specific stimulation was detected by using a cyto-pathic effect reduction assay and splenocytes proliferation of immunized mice in response to antigen restimulation was measured by MTT pTB30m induced higher titer specific antibody against Ag85B in immunized mice and protective cell-mediated immunity,characterized by splenocytes which upon antigen-specific stimulation secreted increased levels of IFNγ and proliferated ┐CLUSION pTB30m should be further tested as DNA vac-cine of TB in preclinical animal models.
0 引言
结核病(tuberculosis,TB)(DRMTB)的流行,以及MTB和HIV双重感染,,,, Ag85B成熟蛋白基因的真核表达载体[1] .在此,我们进一步研究了该重组载体在体外的表达和在小鼠体内诱生体液免疫和细胞免疫应答的能力.
1 材料和方法