中文摘要：

目的： 构建携带中国株HIV-1 gp120基因并可感染小鼠骨髓来源巨噬细胞（BMM）的重组腺病毒。方法： 利用AdMax腺病毒构建系统,以双质粒共转染人胚肾转化细胞293Ad5＋细胞,完成重组腺病毒载体包装,并进一步扩增和纯化。通过ELISA测定gp120的蛋白产量,以确认目的基因重组与表达成功。以Karber法对病毒进 行TCID50滴度测定,利用BMM进行感染复数（MOI）流式细胞术测定,并利用荧光显微镜观察细胞活化形态。结果： 成功构建AdMax-HIV-1 gp120（简称Ad-gp120）及其对照病毒（Ad-GFP）,其滴度分别为108.3和108.1 TCID50/mL。这些病毒可感染BMM,MOI测定结果表明2种重组腺病毒感染BMM和293Ad5＋细胞的能力接近,验证了上述TCID50滴度结果。Ad-gp120可在293Ad5＋细胞中表达gp120蛋白,并可感染和诱导BMM相关形态改变。结论： 成功构建Ad-gp120,其感染可致BMM出现形态发生改变。

英文摘要：

AIM： To construct a recombinant adenovirus carrying gp120 gene of Chinese HIV-1 strain,which can infect mouse bone marrow-derived macrophages （BMM）. METHODS： Co-transfection of shuttle and backbone plasmids of AdMax system into 293Ad5＋ cells was performed, followed by viral packaging, propagation and purification. These viruses were subject to Karber TCID50 titration. The expression of gp120 protein in 293Ad5＋ cells was determined by ELISA. The viral titration was validated by a multiplicity of infection （MOI） test with BMM. RESULTS： The titers of the outcome viruses, including AdMax-HIV-1 gp120 （Ad-gp120） and its vector control Ad-GFP, were 108.3 and 108.1 TCID50/mL, respectively. Both recombinant adenoviruses infected BMM with similar capacity of 293Ad5＋ cell infection, which validated the TCID50 titration.The gp120 protein was positive in 293Ad5＋ cell lysates. BMM activation was observed morphologically after Ad-gp120 infection as compared with Ad-GFP-infected cells. CONCLUSION： Functional adenovirus containing HIV-1 gp120 of prevalent strains in China was successfully constructed. Infection of Ad-gp120 causes BMM activation.