title: Quantifying susceptibility of CD4+ stem memory T-cells to infection by laboratory adapted and clinical HIV-1 strains.
abstract: CD4+ T cells are principal targets for human immunodeficiency virus type 1 (HIV-1) infection. CD4+ T cell subsets are heterogeneous cell populations, divided by functional and phenotypic differences into naïve and memory T cells. The memory CD4+ T cells are further segregated into central, effector and transitional memory cell subsets by functional, phenotypic and homeostatic characteristics. Defining the distribution of HIV-1 infection in different T cell subsets is important, as this can play a role in determining the size and composition of the viral reservoir. Both central memory and transitional memory CD4+ T cells have been described as long-lived viral reservoirs for HIV. Recently, the newly described stem memory T cell subset has also been implicated as a long-lived HIV reservoir. Using green fluorescent protein (GFP) reporter strains of HIV-1 and multi parameter flow cytometry, we developed an assay to simultaneously quantify the susceptibility of stem memory (TSCM), central memory, effector memory, transitional memory and naïve CD4+ T cell subsets, to HIV-1 infection in vitro. We show that TSCM are susceptible to infection with laboratory adapted and clinical HIV-1 strains. Our system facilitates the quantitation of HIV-1 infection in alternative T cell subsets by CCR5- and CXCR4-using viruses across different HIV-1 subtypes, and will be useful for studies of HIV-1 pathogenesis and viral reservoirs.
authors: Center for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia. jflynn@burnet.edu.au.
authors: Jacqueline K
authors: JK
authors: Flynn
authors: Alfred Medical Research and Education Precinct and Burnet Institute Flow Cytometry Core Facility, Melbourne, Victoria 3004, Australia. paukovic@burnet.edu.au.
authors: Geza
authors: G
authors: Paukovics
authors: Center for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia. Kieran@burnet.edu.au.
authors: Kieran
authors: K
authors: Cashin
authors: Center for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia. katharinab@burnet.edu.au.
authors: Katharina
authors: K
authors: Borm
authors: Center for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia. amellett@burnet.edu.au.
authors: Anne
authors: A
authors: Ellett
authors: Center for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia. mroche@burnet.edu.au.
authors: Michael
authors: M
authors: Roche
authors: Department of Biomedicine, Aarhus University, Aarhus 237551, Denmark. mrj@immunology.au.dk.
authors: Martin R
authors: MR
authors: Jakobsen
authors: Center for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia. churchil@burnet.edu.au.
authors: Melissa J
authors: MJ
authors: Churchill
authors: Center for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia. gorry@burnet.edu.au.
authors: Paul R
authors: PR
authors: Gorry
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D005434:Flow Cytometry / Q000379:methods; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000254:growth & development* / Q000302:isolation & purification; D006801:Humans; D013194:Staining and Labeling; D016176:T-Lymphocyte Subsets / Q000821:virology*; D014773:Virology / Q000379:methods*
chemical_list: D049452:Green Fluorescent Proteins