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: 4.65). 11/2002; 76(19):9868-76. DOI: 10.1128/JVI.76.19.9868-9876.2002
Source: PubMed

ABSTRACT 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.

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    • "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. "
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    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 · 3.28 Impact Factor
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    • "Infection by both cell-free and cell-associated virus has been observed in female macaques infected with simian immunodeficiency and chimeric viruses (SIV/SHIV) (Gupta et al., 2002; Kaizu et al., 2006; Khanna et al., 2002; Salle et al., 2010; Zhu et al., 1996), mice infected with HIV (Khanna et al., 2002), and indirectly in humans through genetic matching of HIV viruses sequenced from acutely infected women and from seminal cells and plasma from their infected partners (Zhu et al., 1996). Human cervical explant studies have also confirmed transmission of cell-free and cell-associated HIV (Gupta et al., 2002). Both forms of HIV are carried by semen and deposited in the vagina during intercourse. "
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    ABSTRACT: A combination of prevention and treatment modalities will be needed to successfully control the global spread of HIV. Microbicides, drug products topically applied to mucosal surfaces to prevent HIV infection, are one of these biomedical interventions that hold great promise. In order to be efficacious, microbicides must overcome several challenges imposed by the mucosal microenvironment they intend to protect and the mischievous human immunodeficiency virus with its enormous capacity to adapt. Recent data, however, supports the establishment of the primo-infection by only a small number of founder viruses, which are highly vulnerable to microbicidal intervention at the initial stages of mucosal invasion. The biological foundation of these challenges and opportunities in microbicide development is explored in this review. This article forms part of a special supplement on presentations covering HIV transmission and microbicides, based on the symposium "Trends in Microbicide Formulations", held on 25 and 26 January 2010, Arlington, VA.
    Antiviral research 12/2010; 88 Suppl 1(supplement 1):S3-9. DOI:10.1016/j.antiviral.2010.09.011 · 3.94 Impact Factor
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    • "Exposure of the cervical explants to a topical formulation of UC-781 resulted in blocking of subsequent HIV-1 viral infection of mucosal tissue and transinfection mediated by migratory cells without toxicity, indicating that UC-781 may be a good candidate microbicide. PMPA {9-[2-(phosphonomethoxy)propyl]adenine}, a nucleotide analogue that blocks viral replication by inhibiting reverse transcriptase , was also shown to be able to inhibit viral transmission across the mucosa (Gupta et al., 2002). These findings support the utility of organ cultures with human mucosal tissues to screen topical microbicides for their ability to block sexual transmission of HIV-1. "
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    ABSTRACT: Researchers are recognizing the limitations of two-dimensional (2D) cell cultures, given the fact that they do not reproduce the morphology and biochemical features that the cells possess in the original tissue. As an alternative, the three-dimensional (3D) cell culture approach offers researchers the possibility to study cell growth and differentiation under conditions that more closely resemble the in vivo situation with regard to cell shape and cellular environment. Currently, 3D culture models are being employed in many areas of biomedical research because they offer a more realistic milieu than 2D cultures. The era of 2D culture techniques is moving towards a new epoch of culture systems in 3D. The present review is focused on topics of research on 3D cell cultures in virology and their use in antiviral drug development.
    Antiviral Research 10/2006; 71(2-3):96-107. DOI:10.1016/j.antiviral.2006.05.023 · 3.94 Impact Factor
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