结核分枝杆菌四价DNA疫苗免疫原性和保护效率研究  

Study on Immunogenicity and Protective Efficacy of Tetravalent Combination M.tuberculosis DNA Vaccinea

作　　者：顾田园[1] 蔡宏[2] 田霞[2] 余大海[2] 朱玉贤[2] 

机构地区：[1]北京科技大学土木与环境工程学院,北京100083 [2]北京大学蛋白质工程及植物基因工程国家重点实验室,北京100871

出　　处：《生物化学与生物物理进展》2005年第4期347-352,共6页Progress In Biochemistry and Biophysics

基　　金：国家高技术"863"计划资助项目(2002AA206411).~~

摘　　要：对结核杆菌四价DNA疫苗的免疫应答和保护效果进行了评价.用编码结核分枝杆菌Ag85B、MPT64、MPT70和PstS-3等4种抗原蛋白的基因分别构建的单价DNA疫苗混合成四价苗免疫小鼠.3次免疫后21天,四种抗原特异性抗体滴度分别达到1:6 400、1:51 200、1:6 400、1:6 400.四种蛋白质均能诱导脾脏细胞产生较高水平的抗原特异性IFN-γ,浓度分别为10 582.14 ng/L、13 635.97 ng/L、14 213.15 ng/L和9 657.35 ng/L.三次免疫后经静脉强毒攻击,四价苗组小鼠肺脏和脾脏的载菌数分别减少到阴性组的1/650和1/130.对肺组织的病理形态特征观察表明,空载体免疫的小鼠肺部严重损伤,肺实质干酪样坏死,坏死结节占肺实质的70%～80%,而四价苗免疫的小鼠,肺组织结构正常,肺泡轮廓清晰.研究首次证实,Ag85B、MPT64、MPT70和PstS-3 4种结核杆菌抗原蛋白编码基因组成的四价DNA疫苗,具有很高的免疫应答水平和保护效率.In order to evaluate the immunogenicity and protective efficacy of tetravalent combination M.tuberculosis DNA vaccine, DNA vaccines encoding Ag85B, MPT64, MPT70 and PstS-3 protein were constructed with eukaryotic expression vector pJW4303. Combination DNA vaccines were inoculated intramuscularly into C57BL/6 mice three times at 3 weeks interval. After 21 days of the third injection, the specific antibody titers against the four antigens were 1:6 400, 1:51 200, 1:6 400 and 1:6 400. Meanwhile the mixed spleen cells could produce the high antigen-specific IFN-gamma level in response to the four antigen proteins, IFN-gamma level of Ag85B, MPT64, MPT70 and PstS-3 reached 10 582.14 ng/L, 13 635.97 ng/L, 14 213.15 ng/L and 9 657.35 ng/L respectively. After the last injection, mice were challenged with M.tuberculosis H37Rv. When compared to negative group, the bacterial counts of lung and spleen from the mice vaccinated with tetravalent combination vaccine were reduced about 650 and 130 folds. Microphotographs showed clearly that lungs of mice vaccinated with combination vaccine were much better protected against Mycobacterium tuberculosis challenge than negative mice. The results showed that tetravalent combination M.tuberculosis DNA vaccine elicited both T-cell and humoral immune response, and protected against tuberculosis effectively.

关 键 词：结核分枝杆菌 抗原蛋白 四价DNA疫苗 免疫原性 保护效率 

分 类 号：S852[农业科学—基础兽医学]