Journal: The Journal of Infectious Diseases
Article Title: Prevotella timonensis Bacteria Associated With Vaginal Dysbiosis Enhance Human Immunodeficiency Virus Type 1 Susceptibility Of Vaginal CD4 + T Cells
doi: 10.1093/infdis/jiae166
Figure Lengend Snippet: Antiretroviral drugs used in pre-exposure prophylaxis (PrEP) and combination antiretroviral therapy (cART) block Prevotella timonensis –enhanced human immunodeficiency virus type 1 (HIV-1) infection in vaginal CD4 + T cells. A–C , CD4 + T cells isolated from phytohemagglutinin (PHA)–stimulated peripheral blood mononuclear cells (PBMCs) ( A and B ) or vaginal epithelium explants ( C ) were stimulated overnight by UV-inactivated bacteria ( Lactobacillus crispatus [LC], Lactobacillus iners [LI], Gardnerella vaginalis [GV], Fannyhessea vaginae [FV], Megasphaera elsdenii [ME], and Prevotella timonensis [PT], all on multiplicity of infection [MOI] 10) and subsequently exposed to HIV-1 (SF162; MOI 0.1) for 3 d. HIV-1 infection was measured by flow cytometry after intracellular staining for HIV-1 capsid p24 and depicted here as % p24 + cells of total cells (CD4 + T cells isolated from PHA-stimulated PBMCs ( A and B ) or CD3 + cells of emigrated fraction (explant model, C ). HIV-1 infection in the presence or absence of the PrEP drug tenofovir (TFV, reverse-transcriptase inhibitor, 50 µM, A , n = 4) or replication inhibitors used in cART ( B , n = 3; C , n = 5); maraviroc (Mar, CCR5 blockage, 30 µM); reverse transcriptase inhibitors zidovudine (ZDV, 20 µM) and lamivudine (3TC, 50 µM); raltegravir (Ral, integrase inhibitor, 100 nM); and indinavir (IDV, protease inhibitor, 5 µM). Symbols represent independent donors, bars represent mean ± standard deviation. * P < .05, ** P < .01, *** P < .001, 2-tailed t test.
Article Snippet: Lactobacillus crispatus (German Collection of Microorganisms and Cell Cultures GmbH [DSMZ]-20584), Lactobacillus iners (DSMZ-13335), Gardnerella vaginalis (DSMZ-4944), Fannyhessea vaginae (DMSZ-15829), Megasphaera elsdenii (DSMZ-20460), Bacteroides fragilis (ATCC-25285), Bacteroides thetaiotaomicron (DSMZ-2079), Escherichia coli (NC0749147), Prevotella amnii (DSMZ-23384), Prevotella bivia (DSMZ-20514), Prevotella copri (or Segatella copri ; DSMZ-18205), Prevotella intermedia (DSMZ-20706), and Prevotella timonensis (or Hoylesella timonensis ; DSMZ-22865) were cultured as recommended by DSMZ.
Techniques: Blocking Assay, Virus, Infection, Isolation, Bacteria, Flow Cytometry, Staining, Reverse Transcription, Protease Inhibitor, Standard Deviation
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![Prevotella timonensis –induced human immunodeficiency virus type 1 (HIV-1) uptake, fusion, and viral replication in vaginal CD4 + T cells. Vaginal epithelium explants ( A ) or CD4 + T cells isolated from phytohemagglutinin-stimulated peripheral blood mononuclear cells ( B–D ) were stimulated overnight by UV-inactivated bacteria ( Lactobacillus crispatus [LC], Lactobacillus iners [LI], <t>Gardnerella</t> <t>vaginalis</t> [GV], Fannyhessea vaginae [FV], Megasphaera elsdenii [ME], and Prevotella timonensis [PT], all on multiplicity of infection [MOI] 10) and subsequently exposed to HIV-1 (SF162; MOI 0.1) for 3 d unless stated differently. A , HIV-1 infection was measured by flow cytometry after intracellular staining for HIV-1 capsid p24 and depicted here as % p24 + cells of CD3 + cells of emigrated fraction (explant model, n = 4–8). B , HIV-1 uptake in CD4 + T cells after 4 h HIV-1 exposure as measured by p24 enzyme-linked immunosorbent assay after trypsin treatment and cell lysis (n = 3). C , Pooled data of β-lactamase activity measured by flow cytometry, representing viral fusion upon 4 h infection with NL4.3BaL-BlaM-Vpr (n = 4). D , De novo HIV-1 replication, determined by detecting green fluorescent protein, after HIV-1 NL4.3eGFP-BaL infection (n = 6–7). Symbols represent independent donors, bars represent mean ± standard deviation. * P < .05, ** P < .01, 2-tailed t test.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_2099/pmc11272099/pmc11272099__jiae166f1.jpg)
![Antiretroviral drugs used in pre-exposure prophylaxis (PrEP) and combination antiretroviral therapy (cART) block Prevotella timonensis –enhanced human immunodeficiency virus type 1 (HIV-1) infection in vaginal CD4 + T cells. A–C , CD4 + T cells isolated from phytohemagglutinin (PHA)–stimulated peripheral blood mononuclear cells (PBMCs) ( A and B ) or vaginal epithelium explants ( C ) were stimulated overnight by UV-inactivated bacteria ( Lactobacillus crispatus [LC], Lactobacillus iners [LI], Gardnerella vaginalis [GV], Fannyhessea vaginae [FV], Megasphaera elsdenii [ME], and Prevotella timonensis [PT], all on multiplicity of infection [MOI] 10) and subsequently exposed to HIV-1 (SF162; MOI 0.1) for 3 d. HIV-1 infection was measured by flow cytometry after intracellular staining for HIV-1 capsid p24 and depicted here as % p24 + cells of total cells (CD4 + T cells isolated from PHA-stimulated PBMCs ( A and B ) or CD3 + cells of emigrated fraction (explant model, C ). HIV-1 infection in the presence or absence of the PrEP drug tenofovir (TFV, reverse-transcriptase inhibitor, 50 µM, A , n = 4) or replication inhibitors used in cART ( B , n = 3; C , n = 5); maraviroc (Mar, CCR5 blockage, 30 µM); reverse transcriptase inhibitors zidovudine (ZDV, 20 µM) and lamivudine (3TC, 50 µM); raltegravir (Ral, integrase inhibitor, 100 nM); and indinavir (IDV, protease inhibitor, 5 µM). Symbols represent independent donors, bars represent mean ± standard deviation. * P < .05, ** P < .01, *** P < .001, 2-tailed t test.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_2099/pmc11272099/pmc11272099__jiae166f2.jpg)