重组BCG疫苗和结核杆菌HSP65 DNA疫苗免疫原性的比较研究

Comparative study on immunogenicity from recombinant BCG to HSP65 DNA Vaccine

目的 研究重组BCG(recombinantBCG ,rBCG)疫苗和人结核杆菌热休克蛋白 (heatshockprotein ,HSP) 6 5DNA疫苗对小鼠免疫应答的影响 ,评价两种疫苗的免疫原性。方法 用rBCG疫苗和HSP6 5DNA疫苗免疫BALB/c小鼠 ,免疫 8周时 ,采血、取腹腔巨噬细胞和脾脏进行实验。以NO释放量检测巨噬细胞吞噬活性 ,以淋巴细胞刺激指数 (SI)反映细胞增殖能力 ,以ELISA试剂盒检测血清及脾淋巴细胞培养上清的IL 2和IFN γ的含量。结果 rBCG疫苗以 10 6CFU/鼠的剂量经皮下免疫小鼠后 ,其脾淋巴细胞的增殖能力明显高于对照组 ,BCG组和HSP6 5DNA疫苗组 (P <0 .0 5 ) ;腹腔巨噬细胞一氧化氮 (NO)的含量和血清IL 2的含量明显高于对照组 (P <0 .0 1) ;血清IFN γ的含量明显高于HSP6 5DNA疫苗组 (P <0 .0 5 ) ;其余测定参数均有升高趋势 ,但差异无显著性 (P >0 .0 5 )。而HSP6 5DNA疫苗以 5 0 μg/鼠的剂量经肌注免疫小鼠后 ,其血清IL 2的含量明显高于对照组 (P <0 .0 1) ;血清IFN γ的含量明显低于rBCG组 (P <0 .0 5 ) ;其余测定参数与其他各组相比差异均无显著性(P >0 .0 5 )。结论 本实验结果显示 ,rBCG疫苗具有良好的免疫原性 ,能明显增强BALB/c小鼠的免疫应答反应。HSP6 5DNA疫苗亦能刺激机体的免疫应答 。

Objective The aim of this study is to evaluate the immunogenicity of recombinant BCG(rBCG) and human Mycobacterium tuberculosis heat shock protein(HSP)65 DNA vaccine. Methods The specific pathogen-free BALB/c mice were immunized with rBCG or HSP65 DNA vaccine by sc or im. The lymphocyte stimulating index(SI), macrophage activity as well as IL-2, IFN-γ levels of the serum and culture supernatant of spleen lymphocyte were tested by the end of 8th week of inoculation. Results After immunization with rBCG vaccine in dose of 10 6 CFU/per capita by sc, the SI was significantly higher than those in the control group, BCG and HSP65 DNA groups ( P <0.05); the macrophage activity and serum IL-2 level were also significantly higher than those in the control group ( P <0.01);the serum IFN-γ level was significantly different from that of HSP65 DNA group ( P <0.05); Compared with each groups. Other parameters were no significantly different( P >0.05). After immunization with HSP65 DNA vaccine in dose of 50μg/per capita by intramuscular injection, the serum IL-2 level was significantly higher than that in the control group ( P <0.01);the SI and serum IFN-γ level were significantly lower than that in the rBCG group( P <0.05);compared with each groups, the macrophage activity and the culture supernatant IL-2 level of spleen lymphocyte were no significantly different( P >0.05). Conclusions The rBCG vaccine may induced stronger immune response in BALB/c mice than did the BCG and HSP65 DNA vaccine, whereas the HSP65 DNA vaccine may induced the immune response of BALB/c mice, but its stimulating effect is lower than that by the rBCG vaccine.