title: Modeling HIV-1 Latency in Primary T Cells Using a Replication-Competent Virus.
abstract: HIV-1 latently infected cells in vivo can be found in extremely low frequencies. Therefore, in vitro cell culture models have been used extensively for the study of HIV-1 latency. Often, these in vitro systems utilize defective viruses. Defective viruses allow for synchronized infections and circumvent the use of antiretrovirals. In addition, replication-defective viruses cause minimal cytopathicity because they fail to spread and usually do not encode env or accessory genes. On the other hand, replication-competent viruses encode all or most viral genes and better recapitulate the nuances of the viral replication cycle. The study of latency with replication-competent viruses requires the use of antiretroviral drugs in culture, and this mirrors the use of antiretroviral treatment (ART) in vivo. We describe a model that utilizes cultured central memory CD4(+) T cells and replication-competent HIV-1. This method generates latently infected cells that can be reactivated using latency reversing agents in the presence of antiretroviral drugs. We also describe a method for the removal of productively infected cells prior to viral reactivation, which takes advantage of the downregulation of CD4 by HIV-1, and the use of a GFP-encoding virus for increased throughput.
authors.affiliation: 1 Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine , Salt Lake City, Utah.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Martins
authors.affiliation: 2 Department of Internal Medicine, HIV Translational Research Unit, Ghent University , Ghent, Belgium .
authors.forename: Pawel
authors.initials: P
authors.lastname: Bonczkowski
authors.affiliation: 3 Division of Infectious Diseases, Department of Medicine, University of Utah School of Medicine , Salt Lake City, Utah.
authors.forename: Adam M
authors.initials: AM
authors.lastname: Spivak
authors.affiliation: 2 Department of Internal Medicine, HIV Translational Research Unit, Ghent University , Ghent, Belgium .
authors.forename: Ward
authors.initials: W
authors.lastname: De Spiegelaere
authors.affiliation: 1 Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine , Salt Lake City, Utah.
authors.forename: Camille L
authors.initials: CL
authors.lastname: Novis
authors.affiliation: 1 Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine , Salt Lake City, Utah.
authors.forename: Ana Beatriz
authors.initials: AB
authors.lastname: DePaula-Silva
authors.affiliation: 2 Department of Internal Medicine, HIV Translational Research Unit, Ghent University , Ghent, Belgium .
authors.forename: Eva
authors.initials: E
authors.lastname: Malatinkova
authors.affiliation: 2 Department of Internal Medicine, HIV Translational Research Unit, Ghent University , Ghent, Belgium .
authors.forename: Wim
authors.initials: W
authors.lastname: Trypsteen
authors.affiliation: 1 Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine , Salt Lake City, Utah.
authors.forename: Alberto
authors.initials: A
authors.lastname: Bosque
authors.affiliation: 2 Department of Internal Medicine, HIV Translational Research Unit, Ghent University , Ghent, Belgium .
authors.forename: Linos
authors.initials: L
authors.lastname: Vanderkerckhove
authors.affiliation: 1 Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine , Salt Lake City, Utah.
authors.forename: Vicente
authors.initials: V
authors.lastname: Planelles
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology*; D002478:Cells, Cultured; D015536:Down-Regulation; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology*; D008954:Models, Biological; D014775:Virus Activation / Q000502:physiology*; D017735:Virus Latency / Q000502:physiology*; D014779:Virus Replication / Q000502:physiology
chemical_list: D049452:Green Fluorescent Proteins