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作 者:和绍清[1] 董承红[1] 孙明[1] 李琦涵[1]
机构地区:[1]中国医学科学院中国协和医科大学医学生物学研究所免疫室,昆明650118
出 处:《上海免疫学杂志》2001年第1期25-27,共3页Shanghai Journal of Immunology
摘 要:本研究是通过细胞免疫检测来研究含有两种不同病毒基因的复合型DNA免疫的效果 ,同时和单一病毒基因的DNA免疫作比较。在本实验中 ,将HBV的S基因和 (或 )HIV 1的gp12 0基因插入到真核表达载体pcDNA3中 ,得到能表达HBsAg和gp12 0融合蛋白的重组体pcDNA S 12 0 ,和能表达HBsAg的重组体pcDNA S。在实验中 ,将重组质粒DNA直接注射到BALB/c小鼠股四头肌内 ,按每只小鼠 10 0 μg/ 10 0 μl的剂量经 3次免疫后 ,在T细胞增殖实验中 ,用HBsAg蛋白刺激体外培养的被免疫小鼠T细胞后 ,出现了明显的T细胞增殖。采用51Cr释放法检测特异性CTL杀伤作用时 ,发现致敏的CTL对HIVgp12 0多肽孵育后的靶细胞P815有明显的杀伤作用。Recent studies support the notion that vaccination with plasmid DNA encording genes for viral or bacterial antigens can elicite both humoral and cellular immune responses in rodents and non human primates,but their major advantage at immunologic level are their capacity to induce antigen specific CD8 + T cell responses In the present study,BALB/c mice were immunilized intramuscularly with recombinant plasmid pcDNA S 120 in which the S gene of HBV and gp120 gene of HIV 1 were cloned into pcDNA3 in fusion form,and also with pcDNA S in which the S gene of HBV had been cloned into pcDNA3 Mice were immunilized 3 times with 100 μg/100 μl per mouse In the T cell proliferation assays,it was showed that T cells proliferated in vitro obviously after stimulation with pcDNA S and pcDNA S 120 The sensitized CTL could specifically kill the target cell P815 which were pulsed by 10 AA peptide of HIV It concludes that cellular immune responses could be elicited in the experimental animals after direct injections of recombinatant plasmid DNA in which the S gene and gp120 gene were cloned in fusion form
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