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HIV-1复合多表位-p24嵌合基因重组鸡痘病毒疫苗的构建及小鼠免疫原性 被引量:1

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摘要 将HIV-1p24基因插入至人工设计的复合多表位基因MEG中,获得嵌合基因MEGp24;将MEGp24插入到转移载体pUTA2复合启动子ATI-P7.5×20下游,构建出鸡痘病毒重组转移质粒;经转染和BrdU加压筛选,以基因组PCR和Western blot法筛选出重组病毒;将重组病毒大量制备并纯化后,分别于0,14,42天肌肉注射免疫BALB/c小鼠;第3次免疫后一周,采集小鼠全血与脾脏,分离血清并无菌制备脾淋巴细胞,ELISA法检测小鼠血清HIV-1抗体、脾细胞培养上清中Th1类细胞因子IFN-α和IL-2的含量,流式细胞仪测定CD4+,CD8+T淋巴细胞亚类数量,乳酸脱氢酶(LDH)释放法检测脾特异性CTL杀伤活性.结果表明,重组病毒刺激小鼠产生HIV-1特异性抗体,出现T细胞亚类数量增加,产生针对HIV-1 H-2d限制性CTL表位的特异性靶细胞杀伤作用,同时免疫小鼠脾细胞在HIV-1抗原肽的刺激下分泌Th1类细胞因子的水平大幅增加.研究提示多表位嵌合基因重组FPV疫苗具有良好的免疫原性,可诱导小鼠产生HIV-1特异性细胞和体液免疫应答.
出处 《中国科学(C辑)》 CSCD 北大核心 2007年第1期65-72,共8页 Science in China(Series C)
基金 国家高技术研究发展计划资助项目("863"项目 批准号:2003AA219051)
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