Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells

Vacharaksa, Anjalee; Asrani, Anil C.; Gebhard, Kristin; Fasching, Claudine E.; Giacaman, Rodrigo A.; Janoff, Edward N.; Ross, Kristin; Herzberg, Mark C.

doi:10.1186/1742-4690-5-66

Cited by 30 publications

(59 citation statements)

References 70 publications

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“…The 50% tissue culture infective dose (TCID 50 ) was determined as previously described (10). Briefly, 4 replicates each of cell-associated virus (starting with 200,000 cells/well) and cell-free virus (starting with concentrated virus) were added to the first column of a 96-well plate.…”

Section: Virusesmentioning

confidence: 99%

“…Forty-eight hours after infection, cells were fixed and stained for ␤-galactosidase expression using X-Gal (5-bromo-4-chloro-3-indolyl-␤-D-galactopyranoside; ␤-galactosidase reporter gene staining kit, Sigma-Aldrich) as a substrate. A positive well was defined as one that contained 2 or more blue cells (10). The TCID 50 was calculated using the Spearman-Kaerber formula (11).…”

Section: Virusesmentioning

confidence: 99%

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Efficiency of Cell-Free and Cell-Associated Virus in Mucosal Transmission of Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus

Kolodkin-Gal

Hulot

Korioth-Schmitz

et al. 2013

J Virol

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Effective strategies are needed to block mucosal transmission of human immunodeficiency virus type 1 (HIV-1). Here, we address a crucial question in HIV-1 pathogenesis: whether infected donor mononuclear cells or cell-free virus plays the more important role in initiating mucosal infection by HIV-1. This distinction is critical, as effective strategies for blocking cell-free and cell-associated virus transmission may be different. We describe a novel ex vivo model system that utilizes sealed human colonic mucosa explants and demonstrate in both the ex vivo model and in vivo using the rectal challenge model in rhesus monkeys that HIV-1-infected lymphocytes can transmit infection across the mucosa more efficiently than cell-free virus. These findings may have significant implications for our understanding of the pathogenesis of mucosal transmission of HIV-1 and for the development of strategies to prevent HIV-1 transmission. N ovel microbicide and vaccine candidates for human immunodeficiency virus type 1 (HIV-1) are being evaluated preclinically for efficacy by assessing their ability to protect nonhuman primates against cell-free simian immunodeficiency virus (SIV) or simian-human immunodeficiency virus (SHIV) challenges. However, it remains unclear whether cell-associated virus (virus-infected donor mononuclear cells), cell-free virus, or both play the most important roles in initiating mucosal infection by HIV-1 (1-5). This distinction is critical, since effective strategies for blocking cell-free and cell-associated virus transmission may be very different (3, 6, 7). We sought to explore early events in mucosal transmission of HIV-1 and SIV by evaluating the relative efficiency of cell-associated and cell-free virus in initiating mucosal infection. To model these infection events, we developed a novel three-dimensional sealed human colonic mucosa explant system. We utilized this system in association with the SIV distal colon in vivo challenge model in rhesus macaques to evaluate the relative efficiency of initiating mucosal infection using cell-associated virus compared to that of initiating mucosal infection using cell-free virus in vivo. MATERIALS AND METHODS Viruses.A replication-competent CC chemokine receptor type 5 (CCR5)-tropic HIV-1 strain expressing green fluorescence protein (GFP) [HIV-1(R5) NL4.3-BaL-GFP] (8) was utilized for human organ infections, and SIVmac251 (9) was utilized for rhesus monkey tissue infections.TCID 50 for cell-associated virus and cell-free virus. The 50% tissue culture infective dose (TCID 50 ) was determined as previously described (10). Briefly, 4 replicates each of cell-associated virus (starting with 200,000 cells/well) and cell-free virus (starting with concentrated virus) were added to the first column of a 96-well plate. Then, 5-fold dilutions were performed for a total of 9 serial dilutions (1:5 to 1:5 9 ). An additional column with no virus or cells added served as a negative control to measure the background. TZM-bl cells (NIH AIDS Research and Reference Reagent Pr...

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Section: Virusesmentioning

confidence: 99%

Section: Virusesmentioning

confidence: 99%

Efficiency of Cell-Free and Cell-Associated Virus in Mucosal Transmission of Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus

Kolodkin-Gal

Hulot

Korioth-Schmitz

et al. 2013

J Virol

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“…[9][10][11][12][13][14][15][16] Other studies revealed that direct contact of HIV-1 with the endothelium, which share tight junction structures in common with the epithelium, [17][18][19] could lead to the disruption of endothelial integrity and subsequent increased viral leakage across the endothelium. HIV-1 gp120 and Tat protein have been associated with the disruption of tight junctions in the endothelium.…”

mentioning

confidence: 99%

HIV-1 impairs human retinal pigment epithelial barrier function: possible association with the pathogenesis of HIV-associated retinopathy

Tan

Duan

Xun

et al. 2014

Laboratory Investigation

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The breakdown of human retinal pigment epithelial (HRPE) barrier is considered as the etiology of retinopathy, which affects the quality of life of HIV/AIDS patients. Here we demonstrate that HIV-1 could directly impair HRPE barrier function, which leads to the translocation of HIV-1 and bacteria. HRPE cells (D407) were grown to form polarized, confluent monolayers and treated with different HIV-1 infectious clones. A significant increase of monolayer permeability, as measured by trans-epithelial electrical resistance (TEER) and apical-basolateral movements of sodium fluorescein, was observed. Disrupted tightness of HRPE barrier was associated with the downregulation of several tight junction proteins in D407 cells, including ZO-1, Occludin, Claudin-1, Claudin-2, Claudin-3, Claudin-4, and Claudin-5, after exposure to HIV-1, without affecting the viability of cells. HIV-1 gp120 was shown to participate in the alteration of barrier properties, as evidenced by decreased TEER and weakened expression of tight junction proteins in D407 monolayers after exposure to pseudotyped HIV-1, UV-inactivated HIV-1, and free gp120, but not to an envelope (Env)-defective mutant of HIV. Furthermore, exposure to HIV-1 particles could induce the release of pro-inflammatory cytokines in D407, including IL-6 and MCP-1, both of which downregulated the expression of ZO-1 in the HRPE barrier. Disrupted HRPE monolayer allowed translocation of HIV-1 and bacteria across the epithelium. Overall, these findings suggest that HIV-1 may exploit its Env glycoprotein to induce an inflammatory state in HRPE cells, which could result in impairment of HRPE monolayer integrity, allowing virus and bacteria existing in ocular fluids to cross the epithelium and penetrate the HRPE barrier. Our study highlights the role of HIV-1 in the pathogenesis of HIV/AIDS-related retinopathy and suggests potential therapeutic targets for this ocular complication.

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“…5,6 Significant clinical exposures would require that HIV-1 survive the antiviral activity of saliva and remain infectious within the epithelial cells. We hypothesized, therefore, that HIV-1 escapes inactivation by saliva via rapid uptake into oral epithelial cells.…”

mentioning

confidence: 99%

“…[4][5][6] In the oral cavity, antigen receptive dendritic cells are remote from the mucosal surface, localized to the basal and suprabasal epithelial layers. 14,15 Unlike the vaginal epithelium, 16 the oral intraepithelial dendritic cells (Langerhans cells) do not appear to sample antigen nor capture HIV-1 at the mucosal surface.…”

mentioning

confidence: 99%

Short Communication: HIV Type 1 Escapes Inactivation by Saliva via Rapid Escape into Oral Epithelial Cells

Dietrich

Gebhard

Fasching

et al. 2012

AIDS Research and Human Retroviruses

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Saliva contains anti-HIV-1 factors, which show unclear efficacy in thwarting mucosal infection. When incubated in fresh, unfractionated whole saliva, infectious HIV-1 IIIb and BaL (X4-and R5-tropic, respectively) persisted from 4 to at least 30 min in a saliva concentration-dependent manner. In salivary supernatant for up to 6 h, both infectious HIV-1 strains ''escaped'' into immortalized oral epithelial cells; infectious BaL showed selectively enhanced escape in the presence of saliva. Fluorescently labeled HIV-1 virus-like particles entered oral epithelial cells within minutes of exposure. Using a previously unrecognized mechanism, therefore, strains of HIV-1 escape inactivation by saliva via rapid uptake into oral epithelial cells.

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Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells

Cited by 30 publications

References 70 publications

Efficiency of Cell-Free and Cell-Associated Virus in Mucosal Transmission of Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus

Efficiency of Cell-Free and Cell-Associated Virus in Mucosal Transmission of Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus

HIV-1 impairs human retinal pigment epithelial barrier function: possible association with the pathogenesis of HIV-associated retinopathy

Short Communication: HIV Type 1 Escapes Inactivation by Saliva via Rapid Escape into Oral Epithelial Cells

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