title: Single-Particle Tracking of Human Immunodeficiency Virus Type 1 Productive Entry into Human Primary Macrophages.
abstract: Macrophages are one of the major targets of human immunodeficiency virus (HIV-1), but the viral entry pathway remains poorly understood in these cells. Noninvasive virus labeling and single-virus tracking are effective tools for studying virus entry. Here, we constructed a quantum dot (QD)-encapsulated infectious HIV-1 particle to track viral entry at a single-particle level in live human primary macrophages. QDs were encapsulated in HIV-1 virions by incorporating viral accessory protein Vpr-conjugated QDs during virus assembly. With the HIV-1 particles encapsulating QDs, we monitored the early phase of viral infection in real time and observed that, during infection, HIV-1 was endocytosed in a clathrin-mediated manner; the particles were translocated into Rab5A-positive endosomes, and the core was released into the cytoplasm by viral envelope-mediated endosomal fusion. Drug inhibition assays verified that endosome fusion contributes to HIV-1 productive infection in primary macrophages. Additionally, we observed that a dynamic actin cytoskeleton is critical for HIV-1 entry and intracellular migration in primary macrophages. HIV-1 dynamics and infection could be blocked by multiple different actin inhibitors. Our study revealed a productive entry pathway in macrophages that requires both endosomal function and actin dynamics, which may assist in the development of inhibitors to block the HIV entry in macrophages.
authors.affiliation: College of Life Science and Technology, Huazhong University of Science and Technology , Wuhan 430074, P.R. China.|State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences , Wuhan 430071, P.R. China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Li
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences , Wuhan 430071, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences , Wuhan 430071, P.R. China.
authors.forename: Wen
authors.initials: W
authors.lastname: Yin
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University , Manassas, Virginia 20110, United States.
authors.forename: Jia
authors.initials: J
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences , Wuhan 430071, P.R. China.
authors.forename: Zhi-Ping
authors.initials: ZP
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences , Wuhan 430071, P.R. China.
authors.forename: Dejun
authors.initials: D
authors.lastname: Zeng
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences , Wuhan 430071, P.R. China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University , Manassas, Virginia 20110, United States.
authors.forename: Yuntao
authors.initials: Y
authors.lastname: Wu
authors.affiliation: National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences , Beijing 100101, P.R. China.
authors.forename: Xian-En
authors.initials: XE
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences , Wuhan 430071, P.R. China.
authors.forename: Zongqiang
authors.identifier: 0000-0003-1564-272X
authors.initials: Z
authors.lastname: Cui
mesh_terms: D002478:Cells, Cultured; D057809:HEK293 Cells; D015497:HIV-1 / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000378:metabolism* / Q000821:virology*; D061848:Optical Imaging; D045663:Quantum Dots / Q000737:chemistry / Q000378:metabolism*; D000072760:Single Molecule Imaging / Q000379:methods*
keywords: actin; endosomal fusion; human primary macrophages; productive entry; quantum dots; single-particle tracking