Li Ya-Jing, ZhuGe Fu-Yan, Zeng Chang-Chun, He Jin-Yang, Tan Ning, Liang Juan
School of Biomedical Technology, Guilin Medical University, No. 109, North 2nd Ring Road, Guilin, 541004, Guangxi, China.
School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, Guangdong, China.
Virus Genes. 2017 Apr;53(2):197-204. doi: 10.1007/s11262-016-1423-x. Epub 2016 Dec 28.
A low-cost rodent model of HIV infection and which presents high application value is an effective tool to investigate HIV infection and pathogenesis. However, development of such a small animal model has been hampered by the unsuitability of rodent cells for HIV-1 replication given that the retrovirus HIV-1 has high selectivity to its host cell. Our study used the mouse leukemia cell lines L615 and L1210 that were induced by murine leukemia virus and transfected with hCD4/CCR5 loaded-lentiviral vector. Lentiviral vectors containing the genes hCD4/CCR5 under the transcriptional control of cytomegalovirus promoter were designed. Transfection efficiencies of human CD4 and CCR5 in L615 and L1210 cells were analyzed by quantitative real-time polymerase chain reaction (RT-PCR) and Western blot assay. Results showed that hCD4 and CCR5 proteins were expressed on the cell surface, demonstrating that the L615 and L1210 cells were humanized and that they possess the characteristics necessary for HIV infection of human host cells. Moreover, the sensitivity of human CD4/CCR5 transgenic mouse cells to HIV infection was confirmed by RT-PCR and ELISA. Mouse leukemia cell lines that could express hCD4 and CCR5 were thus established to facilitate normal entry of HIV-1 so that a human CD4/CCR5 transgenic mice cell model can be used to investigate the transmission and pathogenesis of HIV/AIDS and potential antiviral drugs against this disease.
一种低成本且具有高应用价值的HIV感染啮齿动物模型是研究HIV感染和发病机制的有效工具。然而，由于逆转录病毒HIV-1对其宿主细胞具有高度选择性，啮齿动物细胞不适用于HIV-1复制，阻碍了这种小动物模型的开发。我们的研究使用了由鼠白血病病毒诱导并转染了负载hCD4/CCR5的慢病毒载体的小鼠白血病细胞系L615和L1210。设计了在巨细胞病毒启动子转录控制下包含hCD4/CCR5基因的慢病毒载体。通过定量实时聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析来分析人CD4和CCR5在L615和L1210细胞中的转染效率。结果表明，hCD4和CCR5蛋白在细胞表面表达，表明L615和L1210细胞已人源化，并且它们具备人类宿主细胞被HIV感染所需的特征。此外，通过RT-PCR和酶联免疫吸附测定（ELISA）证实了人CD4/CCR5转基因小鼠细胞对HIV感染的敏感性。由此建立了能够表达hCD4和CCR5的小鼠白血病细胞系，以促进HIV-1的正常进入，从而使人CD4/CCR5转基因小鼠细胞模型可用于研究HIV/AIDS的传播和发病机制以及针对该疾病的潜在抗病毒药物。
