摘要
目的探讨结核分枝杆菌复苏因子D(RpfD)-DNA疫苗在BALB/c小鼠抗H37Rv感染中产生的免疫原性。方法将90只SPF级BALB/c小鼠随机分为3组(每组30只),即RpfD疫苗免疫组、BCG免疫组、生理盐水对照组,分别以RpfD-DNA疫苗、BCG和生理盐水于0、3、6周共3次免疫小鼠,于第9周用结核分枝杆菌气雾感染小鼠。于第17周采集小鼠血清,应用ELISA方法检测RpfD抗体效价、IFNγ水平,ELISA检测脾淋巴细胞培养上清诱导的IFNγ、IL-12水平,采用CCK-8检测淋巴细胞增殖指数,LDH方法检测淋巴细胞CTL杀伤效应,并对小鼠肺组织菌荷量计数。结果 RpfD疫苗免疫组小鼠血清RpfD抗体水平和IFNγ水平均显著高于生理盐水组对照组及BCG组(P<0.05或P<0.01)。RpfD体外刺激培养小鼠脾淋巴细胞释放IFNγ水平RpfD疫苗免疫组显著高于其他组(P<0.05);IL-12水平3组间差异无统计学意义(P>0.05)。RpfD疫苗免疫组淋巴细胞增殖指数和脾特异性CTL杀伤性显著高于其他组(P<0.05)。RpfD疫苗免疫组小鼠肺组织结核分枝杆菌CFU显著低于其他组(P<0.05)。结论 RpfD-DNA疫苗可诱导小鼠产生免疫应答,具有免疫原性。
Objective To investigate the immunogenicity resulting from a DNA vaccine of resuscitation-promoting factor D of Mycobacterium tuberculosis（RpfD） against H37Rv infection in BALB/c mice.Methods First,90 SPF-grade BALB/c mice were randomly divided into 3 groups（n=30）,an RpfD DNA vaccine group,a BCG vaccine group,and a normal saline control group.The RpfD DNA vaccine,BCG,and normal saline were given to mice at 0,3,and 6 weeks.In the ninth week,mice were exposed to aerosolized M.tuberculosis.In the seventeenth week,（1） ELISA was used to test mouse serum and measure the RpfD antibody titer and IFNγ levels,（2） ELISA was used to measure serum-induced IFNγ and IL-12 levels in cultured splenic lymphocytes,（3） CCK-8 was used to determine the lymphocyte proliferation index of murine splenic lymphocytes and LDH was used to measure the CTL activity of lymphocytes,and（4） the bacterial load in mouse lung tissue was determined.Results The RpfD vaccine group had significantly higher levels of RpfD antibodies in serum compared to other groups（P0.01）.The RpfD vaccine group also had significantly higher IFNγ levels than the normal saline group（P0.01） and the BCG group（P0.05）.The RpfD vaccine group had significantly higher IFNγ levels released by cultured murine splenic lymphocytes in response to in vitro stimulation with RpfD protein compared to other groups（P0.05）.Differences in IL-12 levels among the 3 groups were not statistically significant（P0.05）.The RpfD vaccine group had significantly higher CCK-8（lymphocyte proliferation index） and LDH（spleen-specific CTL activity） results compared to other groups（P0.05）.The RpfD vaccine group had a significantly fewer M.tuberculosis CFUs in lung tissue compared to the other groups（P0.05）.Conclusion The RpfD-DNA vaccine induced an immune response in mice that provided immunogenicity.
出处
《中国病原生物学杂志》
CSCD
北大核心
2012年第10期739-742,共4页
Journal of Pathogen Biology
