小鼠对HIV-2嵌合结构基因重组鸡痘病毒的免疫反应  

Cellular and humoral immne reponses induced by HIV-2 recombinant fowlpox virus on mice

作　　者：李子健[2] 张立树[1] 金宁一[1] 江文正[3] 李昌[1] 

机构地区：[1]北京大学第三医院血管医学研究所,北京100083 [2]中国人民解放军军事医学科学院军事兽医研究所全军基因工程重点实验室,长春130062 [3]华东师范大学生命科学学院,上海200026

出　　处：《高技术通讯》2006年第7期730-734,共5页Chinese High Technology Letters

基　　金：863计划（2003AA219051）、吉林省科技发展计划（20030550-1）资助项目.

摘　　要：将HIV-2嵌合结构基因gag-gp105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pA-gg;经转染和BrdU加压筛选,以基因组PCR和Western blot法鉴定重组病毒;将获得的重组病毒大量制备并纯化后,肌肉注射免疫BALB/c小鼠,ELISA法检测小鼠血清HIV-2抗体,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,成功筛选出一株可稳定表达HIV-2嵌合蛋白Gag-gp105的重组鸡痘病毒rFPV-gg;该重组病毒免疫组小鼠的血清出现HIV-2抗体,脾T细胞亚类数量增加,并产生针对HIV-2靶细胞的特异性CTL杀伤活性.本研究提示,HIV-2 gag-gp105重组病毒能诱导小鼠产生特异性细胞和体液免疫应答.A fowlpox virus （FPV） transferring vector pUTA2-gag-gp105 was constructed by inserting HIV-2 gag-gpl05 chimeric gene to the downstream of a synthetic complex promoter ATI-p7.5× 20 of vector pUTA2. Transfection was then carried out, and recombinant FPV （rFPV） was screened by 5＇-bromo-deoxym＇idine （BrdU）, genome PCR and Western blot detection. Hereditary stability of the rFPV was detected. BALB/c mice were immunized with rFPV by muscular injection. Anti-HIV-2 antibody, CD^4＋ and CD^8＋ T-cell count and specific target-killing activity of spleen CTL in immunized mice were analyzed by ELISA, FACS and LDH release assay, respectively. The results showed that a recombinant virus rFPV- gg which could stablely express HIV-2 chimeric protein gag-gpl05 was obtained. The HIV-2 specific antibody was detect- ed from the immunized BALB/c mice. The numbers of CD^4＋ and CD^8＋ subgroup of spleen T lymphocyte were higher in immunization group than those in controls. HIV-2-Specific target-killing activity of spleen CTL was observed in immunized mice. It was concluded that the rFPV-gg may elicit specific celluar and humoral immune reactions in mice.

关 键 词：人免疫缺陷病毒Ⅱ型(HIV-2) 结构基因 重组 鸡痘病毒 免疫反应 

分 类 号：Q95-331[生物学—动物学] S852.65[农业科学—基础兽医学]