title: Establishment of a cell line with stable expression of mCherry-EGFP tandem fluorescent-tagged LC3B for studying the impact of HIV-1 infection on autophagic flux.
abstract: Increasing evidence indicates that HIV-1 infection has an impact on cell autophagy, and a susceptible cell line is required for studying the relationship of HIV-1 with autophagy. However, there is limited information on the optimal cell line to evaluate the changes of autophagy affected by HIV infection. In this study cell line TZM-tfLC3B was constructed to express mCherry-EGFP tandem fluorescent tagged LC3B (tfLC3B) by stable transfection of tfLC3B as well as allowing X4/R5 tropic HIV-1 replication. The monitoring of autophagic flux in TZM-tfLC3B was achieved by observing fluorescent puncta. HIV-1 virus-like particles lacking replicative nucleic acid could induce autophagy in TZM-tfLC3B in an envelope glycoprotein dependent manner. These data suggest that TZM-tfLC3B will be a useful tool for studying the HIV-1-induced autophagy modulation of host cells.
authors.affiliation: Laboratory of Medical Molecular Virology, School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, China. Electronic address: Lbin1984@gmail.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.affiliation: Laboratory of Medical Molecular Virology, School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, China. Electronic address: changliu@nankai.edu.cn.
authors.forename: Chang
authors.initials: C
authors.lastname: Liu
authors.affiliation: Laboratory of Medical Molecular Virology, School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, China. Electronic address: zxc832186@163.com.
authors.forename: Xuechao
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Laboratory of Medical Molecular Virology, School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, China. Electronic address: shenwenyuan66@126.com.
authors.forename: Wenyuan
authors.initials: W
authors.lastname: Shen
authors.affiliation: Laboratory of Medical Molecular Virology, School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, China. Electronic address: 775133958@qq.com.
authors.forename: Lingyu
authors.initials: L
authors.lastname: Qian
authors.affiliation: State Key Laboratory of Biotherapy/Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, No. 17, Section 3, Renmin South Road, Chengdu, China. Electronic address: yuquawei@vip.sina.com.
authors.forename: Yuquan
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Laboratory of Medical Molecular Virology, School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, China. Electronic address: kongxh@nankai.edu.cn.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Kong
mesh_terms: D001343:Autophagy; D002460:Cell Line; D003584:Cytological Techniques / Q000379:methods*; D015497:HIV-1 / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods*; D014773:Virology / Q000379:methods
chemical_list: D008164:Luminescent Proteins; C502070:MAP1LC3B protein, human; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins
keywords: Autophagy; LC3B; Stable transfection; TZM-bl; Virus-like particle