title: Establishment of mouse leukemia cell lines expressing human CD4/CCR5 using lentiviral vectors.
abstract: 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.
authors.affiliation: School of Biomedical Technology, Guilin Medical University, No. 109, North 2nd Ring Road, Guilin, 541004, Guangxi, China.
authors.forename: Ya-Jing
authors.initials: YJ
authors.lastname: Li
authors.affiliation: School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, Guangdong, China.
authors.forename: Fu-Yan
authors.initials: FY
authors.lastname: ZhuGe
authors.affiliation: School of Biomedical Technology, Guilin Medical University, No. 109, North 2nd Ring Road, Guilin, 541004, Guangxi, China. 373821547@qq.com.|Affiliated Longhua Central Hospital, Guangdong Medical University, Shenzhen, 541004, Guangdong, China. 373821547@qq.com.
authors.forename: Chang-Chun
authors.initials: CC
authors.lastname: Zeng
authors.affiliation: Tropical Medicine Institute, Guangzhou University of Chinese Medicine, No. 12, Jichang Road, Guangzhou, 510405, Guangdong, China. 303877469@qq.com.
authors.forename: Jin-Yang
authors.initials: JY
authors.lastname: He
authors.affiliation: Guangxi Key Laboratory of Molecular Medicine in Liver Injury and Repair, Guilin Medical University, Guilin, 541004, Guangxi, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Tan
authors.affiliation: School of Biomedical Technology, Guilin Medical University, No. 109, North 2nd Ring Road, Guilin, 541004, Guangxi, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Liang
mesh_terms: D000818:Animals; D015704:CD4 Antigens / Q000096:biosynthesis* / Q000235:genetics; D004195:Disease Models, Animal; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D015658:HIV Infections / Q000188:drug therapy / Q000235:genetics* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000472:pathogenicity; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D009052:Leukemia Virus, Murine / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D019713:Receptors, CCR5 / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: C584933:CCR5 protein, human; D015704:CD4 Antigens; D019713:Receptors, CCR5
keywords: CCR5; CD4; Gene transfer; HIV-1; Lentiviral vectors; Mouse leukemia cell lines (L615 and L1210)