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目的研究结核分枝杆菌重组Ag85A(rAg85A)和/或Ag85B(rAg85B)蛋白与母牛分枝杆菌菌苗(Mycobacteri-um vaccae,MV)联合免疫的免疫原性。方法 60只雌性BALB/c小鼠随机分为下列6组:(1)阴性对照组:磷酸盐缓冲液(PBS)组,(2)阳性对照组:卡介苗(BCG)组,(3)阳性对照组:MV组,(4)实验组:rAg85A-MV组,(5)实验组:rAg85B-MV组及(6)实验组:rAg85A-rAg85B-MV组。每2周免疫1次,共免疫3次。末次免疫结束后第14天杀鼠,用ELISA法检测血清中IgGI、gG1和IgG2a水平;ELISPOT检测脾脏分泌γ干扰素(IFN-γ)的T淋巴细胞斑点数;流式细胞术检测全血单个核细胞内Th1和Th2百分比。结果三个实验组小鼠血清抗体水平均明显高于三个对照组(P<0.001);三个实验组和两个阳性对照组小鼠的T淋巴细胞斑点数均明显高于阴性对照组(P<0.05);rAg85A-rAg85B-MV组小鼠Th1百分比明显高于其余五组(P<0.001),三个实验组Th1/Th2比值均明显高于三个对照组(P<0.05)。结论结核分枝杆菌rAg85A和rAg85B蛋白均具有很强的免疫原性,与MV联合免疫可增强其Th1型细胞免疫应答。
Objective To study the immunogenicity of combined immunization with M. tuberculosis recombinant Ag85A (rAg85A) and / or Ag85B (rAg85B) protein and Mycobacterium vaccae (MV). Methods Sixty female BALB / c mice were randomly divided into the following 6 groups: (1) Negative control group: PBS group; (2) Positive control group: BCG group; (3) Positive control group Group: MV group, (4) experimental group: rAg85A-MV group, (5) experimental group: rAg85B-MV group and (6) experimental group: rAg85A-rAg85B-MV group. Immunization every 2 weeks, a total of 3 immunizations. The mice were sacrificed on the 14th day after the last immunization and the levels of IgGI, gG1 and IgG2a in the serum were detected by ELISA. The T lymphocytes of IFN-γ secreted by the spleen were detected by ELISPOT. Percentage of Th1 and Th2 in nucleated cells. Results The levels of serum antibody in the three experimental groups were significantly higher than those in the three control groups (P <0.001). The numbers of T lymphocytes in three experimental groups and two positive control groups were significantly higher than those in the negative control group P <0.05). The Th1 percentage of mice in rAg85A-rAg85B-MV group was significantly higher than that of the other five groups (P <0.001). Th1 / Th2 ratios in three experimental groups were significantly higher than those in the three control groups (P <0.05). Conclusion Both Mycobacterium tuberculosis rAg85A and rAg85B proteins have strong immunogenicity. Combined immunization with MV can enhance their Th1-type cellular immune response.