Despite extensive research efforts, a preventive human immunodeficiency virus (HIV) vaccine remains one of the major challenges in the field of AIDS research. Experimental strategies which have been proven successful ...Despite extensive research efforts, a preventive human immunodeficiency virus (HIV) vaccine remains one of the major challenges in the field of AIDS research. Experimental strategies which have been proven successful for other viral vaccines are not enough to tackle HIV-1 and new approaches to design effective preventive AIDS vaccines are of utmost importance. Due to enormous diversity among global circulating HIV strains, an effective HIV vaccine must elicit broadly protective antibodies based responses;therefore discovering new broadly neutralizing antibodies (bNAbs) against HIV has become major focus in HIV vaccine research. However further understanding of the viral targets of such antibodies and mechanisms of action of bNAbs is required for advancement of HIV vaccine research. This technical note discusses our current knowledge on the bNAbs and immunoprophylaxis using viral vectors with their relevance in designing of new candidates to HIV-1 vaccines.展开更多
To construct the eukaryotic expression vector of HIV-1 gp120 gene and observe its expression in vitro, the recombinant expression vector pVAX1GP120 was constructed by inserting the gp120 gene into the eukaryotic expre...To construct the eukaryotic expression vector of HIV-1 gp120 gene and observe its expression in vitro, the recombinant expression vector pVAX1GP120 was constructed by inserting the gp120 gene into the eukaryotic expression vector pVAX1. The pVAX1GP120 was transfected into Vero cells by lipofectamine and the expressed product was detected by indirect immunofluore- scence. Restriction enzymes digestion analysis and sequencing results revealed that the recombinant expression vector pVAX1GP120 has been constructed successfully. The indirect immunofluorescence result showed green fluorescence on the membrane of transfected cells. The constructed eukaryotic expression vector of HIV-1 gp120 can be expressed in vitro, which lay the foundation for the further study of HIV-1 DNA vaccine.展开更多
目的:构建HIV 1辅受体的配体-趋化因子RAN TES和SDF 1的逆转录病毒载体,并观察在NIH3T3细胞的表 达.方法:从重组质粒pCMV R K S K中获取RANTES KDEL SDF KDEL基因片段,构建RANTES和SDF 1的逆转录病毒 载体pLNCX R K S K,酶切鉴定并测...目的:构建HIV 1辅受体的配体-趋化因子RAN TES和SDF 1的逆转录病毒载体,并观察在NIH3T3细胞的表 达.方法:从重组质粒pCMV R K S K中获取RANTES KDEL SDF KDEL基因片段,构建RANTES和SDF 1的逆转录病毒 载体pLNCX R K S K,酶切鉴定并测序.采用标准的磷酸钙共 沉淀法转染包装细胞Bing,G418筛选克隆细胞,制备重组病 毒液,继之感染NIH3T3细胞,计算病毒滴度.间接免疫荧光 检测NIH3T3细胞中RANTES、SDF 1的表达.结果:pLNCX R K S K逆转录病毒载体酶切和测序结果与预期一致,筛选 出抗性克隆,制备了高滴度的重组病毒液,间接免疫荧光证实 RANTES和SDF 1可表达于转染细胞.结论:HIV 1辅受体的 配体、趋化因子RANTES和SDF 1的逆转录病毒载体构建成 功,重组病毒可以感染NIH3T3细胞,为下一步HIV 1感染实 验打下了基础.展开更多
文摘Despite extensive research efforts, a preventive human immunodeficiency virus (HIV) vaccine remains one of the major challenges in the field of AIDS research. Experimental strategies which have been proven successful for other viral vaccines are not enough to tackle HIV-1 and new approaches to design effective preventive AIDS vaccines are of utmost importance. Due to enormous diversity among global circulating HIV strains, an effective HIV vaccine must elicit broadly protective antibodies based responses;therefore discovering new broadly neutralizing antibodies (bNAbs) against HIV has become major focus in HIV vaccine research. However further understanding of the viral targets of such antibodies and mechanisms of action of bNAbs is required for advancement of HIV vaccine research. This technical note discusses our current knowledge on the bNAbs and immunoprophylaxis using viral vectors with their relevance in designing of new candidates to HIV-1 vaccines.
文摘To construct the eukaryotic expression vector of HIV-1 gp120 gene and observe its expression in vitro, the recombinant expression vector pVAX1GP120 was constructed by inserting the gp120 gene into the eukaryotic expression vector pVAX1. The pVAX1GP120 was transfected into Vero cells by lipofectamine and the expressed product was detected by indirect immunofluore- scence. Restriction enzymes digestion analysis and sequencing results revealed that the recombinant expression vector pVAX1GP120 has been constructed successfully. The indirect immunofluorescence result showed green fluorescence on the membrane of transfected cells. The constructed eukaryotic expression vector of HIV-1 gp120 can be expressed in vitro, which lay the foundation for the further study of HIV-1 DNA vaccine.
文摘目的:构建HIV 1辅受体的配体-趋化因子RAN TES和SDF 1的逆转录病毒载体,并观察在NIH3T3细胞的表 达.方法:从重组质粒pCMV R K S K中获取RANTES KDEL SDF KDEL基因片段,构建RANTES和SDF 1的逆转录病毒 载体pLNCX R K S K,酶切鉴定并测序.采用标准的磷酸钙共 沉淀法转染包装细胞Bing,G418筛选克隆细胞,制备重组病 毒液,继之感染NIH3T3细胞,计算病毒滴度.间接免疫荧光 检测NIH3T3细胞中RANTES、SDF 1的表达.结果:pLNCX R K S K逆转录病毒载体酶切和测序结果与预期一致,筛选 出抗性克隆,制备了高滴度的重组病毒液,间接免疫荧光证实 RANTES和SDF 1可表达于转染细胞.结论:HIV 1辅受体的 配体、趋化因子RANTES和SDF 1的逆转录病毒载体构建成 功,重组病毒可以感染NIH3T3细胞,为下一步HIV 1感染实 验打下了基础.
