Memory CD4+ T Cells Are the Earliest Detectable Human Immunodeficiency Virus Type 1 (HIV-1)-Infected Cells in the Female Genital Mucosal Tissue during HIV-1 Transmission in an Organ Culture System

Department of Infectious Diseases and Microbiology, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
Journal of Virology (Impact Factor: 4.44). 11/2002; 76(19):9868-76. DOI: 10.1128/JVI.76.19.9868-9876.2002
Source: PubMed


The virologic and cellular factors that are involved in transmission of human immunodeficiency virus type 1 (HIV-1) across the female genital tissue are poorly understood. We have recently developed a human cervical tissue-derived organ culture model to study heterosexual transmission of HIV-1 that mimics the in vivo situation. Using this model we investigated the role of phenotypic characteristics of HIV-1 and identified the cell types that are first infected during transmission. Our data indicate that the cell-free R5 HIV-1 was more efficiently transmitted than cell-free X4 HIV-1. Cell-free and cell-associated HIV-1 had comparable transmission efficiency regardless of whether the virus was of R5 or X4 type. We have demonstrated that memory CD4(+) T cells and not Langerhans cells were the first HIV-1 RNA-positive cells detected at the epithelial-submucosal junction 6 h after virus exposure. Multicolor laser confocal microscopy demonstrated a globular distribution of HIV-1 gag-pol mRNA in the cytoplasm, and the distribution of CD4 and the CD45RO isoform was irregular on the cellular membrane. At 96 h postinoculation, in addition to memory CD4(+) T cells, HIV-1 RNA-positive Langerhans cells and macrophages were also detected. The identification of CD4(+) T cells in the tissue at 6 h was confirmed by flow cytometric simultaneous immunophenotyping and ultrasensitive fluorescence in situ hybridization assay on immune cells isolated from disaggregated tissue. Furthermore, PMPA [9-[2-(phosphonomethoxy)propyl] adenine], an antiretroviral compound, and UC781, a microbicide, inhibited HIV-1 transmission across the mucosa, indicating the utility of the organ culture to screen topical microbicides for their ability to block sexual transmission of HIV-1.

Download full-text


Available from: Simon C Watkins, Oct 02, 2015
  • Source
    • "These cells are also present in the lamina propria, subepithelial stroma, and the epithelium (Edwards and Morris, 1985; Hickey et al., 2011; Johansson et al., 1999). Findings using human vaginal explants show that CD4 þ T cells are relevant to HIV-1 transmission because HIV-1 productively infects both activated and resting CD4 þ T cells in the genital mucosa (Gupta et al., 2002; Hladik et al., 2007; Zhang et al., 1999). Whether DCs, Langerhans cells, and monocytes present in the cervicovaginal Fig. 5. Human semen-derived exosomes decrease murine susceptibility to LP-BM5 mAIDS virus infection: (A) schematic representation of experimental procedure. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Exosomes are membranous extracellular nanovesicles secreted by diverse cell types. Exosomes from healthy human semen have been shown to inhibit HIV-1 replication and to impair progeny virus infectivity. In this study, we examined the ability of healthy human semen exosomes to restrict HIV-1 and LP-BM5 murine AIDS virus transmission in three different model systems. We show that vaginal cells internalize exosomes with concomitant transfer of functional mRNA. Semen exosomes blocked the spread of HIV-1 from vaginal epithelial cells to target cells in our cell-to-cell infection model and suppressed transmission of HIV-1 across the vaginal epithelial barrier in our trans-well model. Our in vivo model shows that human semen exosomes restrict intravaginal transmission and propagation of murine AIDS virus. Our study highlights an antiretroviral role for semen exosomes that may be harnessed for the development of novel therapeutic strategies to combat HIV-1 transmission. Copyright © 2015 Elsevier Inc. All rights reserved.
    Virology 08/2015; 482. DOI:10.1016/j.virol.2015.03.040 · 3.32 Impact Factor
  • Source
    • "In this study we focused on the infection of cervical T cells, which have also been reported to be the earliest detectable infected cells in human genital mucosa ex-vivo [17]. However, according to some reports dendritic cells (DCs) and macrophages also may play an important role in the early events of HIV infection. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Recently, it was found that 80% of sexual HIV-1 transmissions are established by a single virion/viral genome. To investigate whether the transmitted/founder (T/F) viruses have specific biological properties favoring sexual transmission, we inoculated human cervical tissue explants with isogenic HIV-1 viruses encoding Env sequences from T/F and control reference (C/R) HIV-1 variants as well as with full length T/F HIV-1 and compared their replication efficiencies, T cell depletion, and the activation status of infected cells. We found that all the HIV-1 variants were capable of transmitting infection to cervical tissue ex vivo and in this system preferentially replicate in activated CD4 T cells and deplete these cells. There was no difference in the biological properties of T/F and C/R HIV-1 variants as evaluated in ex vivo cervical tissue.
    PLoS ONE 12/2012; 7(12):e50839. DOI:10.1371/journal.pone.0050839 · 3.23 Impact Factor
  • Source
    • "The latter occurs via induction of inflammatory cytokines and subsequent downregulation of junction genes, which facilitate transport of HIV-1 across the epithelial barrier. However, DCs or LCs are proposed as initial target cells of HIV-1 that mediate transfer of the virus to CD4+ T cells in the draining lymph nodes, which is supported by both ex vivo and in vivo studies, (Spira et al., 1996; Hu et al., 2000; Gupta et al., 2002; Veazey et al., 2003; Hladik et al., 2007). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Dendritic cells (DCs), Langerhans cells (LCs), and macrophages are innate immune cells that reside in genital and intestinal mucosal tissues susceptible to HIV-1 infection. These innate cells play distinct roles in initiation of HIV-1 infection and induction of anti-viral immunity. DCs are potent migratory cells that capture HIV-1 and transfer virus to CD4(+) T cells in the lymph nodes, whereas LCs have a protective anti-viral function, and macrophages function as viral reservoirs since they produce viruses over prolonged times. These differences are due to the different immune functions of these cells partly dependent on the expression of specific pattern recognition receptors. Expression of Toll-like receptors, C-type lectin receptors, and cell-specific machinery for antigen uptake and processing strongly influence the outcome of virus interactions.
    Frontiers in Immunology 03/2012; 3:59. DOI:10.3389/fimmu.2012.00059
Show more