【摘要】 目的　研究结核分支杆菌 (Mycobacteriumtuberculosis,MTB)MPT64质粒DNA的免疫原性和保护效力。方法　分别用生理盐水 (A)、载体质粒 (B)、卡介苗 (C)和MPT64重组质粒 (D)免疫小鼠 ,通过ELISA方法检测小鼠血清中抗MPT64抗体 ,免疫 8周后以MTB腹腔内攻击小鼠 ,攻击 4或 9周后 ,分别取肺、肝和脾脏观察病理改变、称取重量、作菌落计数及脾淋巴细胞转化试验。结果　D组1 0只小鼠中只有 4只血清中检测出抗MPT64抗体。MTB攻击 4周后 ,D组鼠脾淋巴细胞转化率显著高于其它 3组 ,A、B、D组肺、肝和脾重量明显高于C组 ,A组脾菌落数较其它 3组明显增多 ,其余脏器菌落数各组无明显区别 ,A、B组可见肺部严重病变 ,C组未见明显病变 ,D组存在个体差异。MTB攻击 9周后 ,A、B组鼠脾淋巴细胞转化率较C组显著降低 ,D组显著增高 ,各组肺、肝和脾重量无显著差别 ,C组肝菌落数明显低于其它 3组 ,其余脏器各组间无显著差别 ,各组鼠肺表面均可见不同程度的病变。所有鼠肝、脾均未见明显病变。结论　MTBMPT64DNA疫苗经肌肉注射免疫小鼠能诱发特异的体液和细胞免疫应答 ,对小鼠抗MTB感染具有一定的免疫保护效力 ,但比卡介苗差更多还原

【Abstract】 Objective To study the immunogenicity and protective efficacy of M.tuberculosis MPT64 plasmid DNA.Methods The mice were immunized with the saline (A),plasmid vector (B), M.bovis BCG (C),and MPT64 plasmid DNA (D),and were then infected by intraperitoneal injection M.tuberculosis strain H 37 Rv.MPT64 specific antibodies were determined by ELISA.Their lungs,livers and spleens were taken and observed their pathological changes,weighted and performed mycobacterial cultures 4 or 9 weeks after infection.Spleen cells were tested for proliferative responses.Results MPT64 specific antibodies increased in 4 of 10 group D mice.The stimulation rates of spleen cells increased significantly in group D mice compared to those in the other three group mice at 4 weeks after infection.The weights of lungs,livers and spleens in group A,B and D mice were higher than those in group C mice.The spleens CFUs in group A were more than those in the other three groups.The CFUs of lungs and livers had no significant difference among these four groups.The serious lesions could be observed in lungs of group A and B,while not in group C.Group D were different individually.The stimulation rates of spleen cells decreased significantly in group A and B mice,and increased significantly in group D mice compared to those in group C mice at 9 weeks after infection.The weights of lungs,livers and spleens had no significant difference among these four groups.The livers CFUs in group C mice were lower than those in the other three groups mice.Some lesions could be observed in lungs,but not in liver and spleen of each group mice.Conclusions M.tuberculosis MPT64 DNA vaccine,which was immunized mice by intramuscle injection,could induce specific humoral and cellular immune response and provide some degree of protection against Mycobacterium tuberculosis challenge.更多还原