The Progestin Medroxyprogesterone Acetate Affects HIV-1 Production in Human Lymphoid Tissue Explants in a Dose-Dependent and Glucocorticoid-like Fashion

The association between the use of the injectable contraceptive depot medroxyprogesterone acetate and HIV-1 susceptibility has been addressed mainly in respect to the changes occurring in the female genital mucosa and blood. However, one of the main sites of HIV-1 pathogenesis is lymphoid organs. To investigate the immunoregulatory effect of medroxyprogesterone acetate (MPA) at this site, human tonsillar tissue explants were infected ex vivo with either a CCR5 (BaL) or CXCR4 (LAI) HIV-1 variant and the release of p24gag and cytokines was measured in culture supernatant. The response to MPA was compared with that elicited by treatment with progesterone (P4) and dexamethasone (DEX), which selectively binds the glucocorticoid receptor, in donor-matched explant cultures. MPA treatment reduced the replication of both tested HIV-1 strains as well as the production of the mediators of inflammation IL-1β, IL-17A and CCL5, but not CCL20, in a similar way to DEX, whereas P4 had no effect on HIV-1 replication. The magnitude of both MPA and DEX-mediated responses was proportional to the length of exposure and/or administered dose. Blockage of the progesterone and glucocorticoid receptors with mifepristone abolished all observed changes in HIV-1 and cytokine production, and was associated with increased IL-22 levels in HIV-infected explants. Our data indicate that elevated doses of MPA may affect the immune responses in lymphoid tissue in a glucocorticoid-like fashion with an immediate impact on local HIV-1 replication.

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Vpu serine 52 dependent counteraction of tetherin is required for HIV-1 replication in macrophages, but not in ex vivo human lymphoid tissue

Retrovirology ◽

10.1186/1742-4690-7-1 ◽

2010 ◽

Vol 7 (1) ◽

Cited By ~ 59

Author(s):

Michael Schindler ◽

Devi Rajan ◽

Carina Banning ◽

Peter Wimmer ◽

Herwig Koppensteiner ◽

...

Keyword(s):

Lymphoid Tissue ◽

Ex Vivo ◽

Human Lymphoid Tissue ◽

Hiv 1

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Human Immunodeficiency Virus Type 1 Variants Resistant to First- and Second-Version Fusion Inhibitors and Cytopathic in Ex Vivo Human Lymphoid Tissue

Journal of Virology ◽

10.1128/jvi.02546-06 ◽

2007 ◽

Vol 81 (12) ◽

pp. 6563-6572 ◽

Cited By ~ 33

Author(s):

Raghavan Chinnadurai ◽

Devi Rajan ◽

Jan Münch ◽

Frank Kirchhoff

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Lymphoid Tissue ◽

Ex Vivo ◽

Fusion Inhibitors ◽

Immunodeficiency Virus ◽

Human Lymphoid Tissue ◽

Hiv 1

ABSTRACT Human immunodeficiency virus type 1 (HIV-1) fusion inhibitors blocking viral entry by binding the gp41 heptad repeat 1 (HR1) region offer great promise for antiretroviral therapy, and the first of these inhibitors, T20 (Fuzeon; enfuvirtide), is successfully used in the clinic. It has been reported previously that changes in the 3-amino-acid GIV motif at positions 36 to 38 of gp41 HR1 mediate resistance to T20 but usually not to second-version fusion inhibitors, such as T1249, which target an overlapping but distinct region in HR1 including a conserved hydrophobic pocket (HP). Based on the common lack of cross-resistance and the difficulty of selecting T1249-resistant HIV-1 variants, it has been suggested that the determinants of resistance to first- and second-version fusion inhibitors may be different. To further assess HIV-1 resistance to fusion inhibitors and to analyze where changes in HR1 are tolerated, we randomized 16 codons in the HR1 region, including those making contact with HR2 codons and/or encoding residues in the GIV motif and the HP. We found that changes only at positions 37I, 38V, and 40Q near the N terminus of HR1 were tolerated. The propagation of randomly gp41-mutated HIV-1 variants in the presence of T1249 allowed the effective selection of highly resistant forms, all containing changes in the IV residues. Overall, the extent of T1249 resistance was inversely correlated to viral fitness and cytopathicity. Notably, one HIV-1 mutant showing ∼10-fold-reduced susceptibility to T1249 inhibition replicated with wild type-like kinetics and caused substantial CD4+-T-cell depletion in ex vivo-infected human lymphoid tissue in the presence and absence of an inhibitor. Taken together, our results show that the GIV motif also plays a key role in resistance to second-version fusion inhibitors and suggest that some resistant HIV-1 variants may be pathogenic in vivo.

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The role of upstream U3 sequences in HIV-1 replication and CD4+ T cell depletion in human lymphoid tissue ex vivo

Virology ◽

10.1016/j.virol.2005.07.023 ◽

2005 ◽

Vol 341 (2) ◽

pp. 313-320 ◽

Cited By ~ 8

Author(s):

Jan Münch ◽

Devi Rajan ◽

Elke Rücker ◽

Steffen Wildum ◽

Nadia Adam ◽

...

Keyword(s):

T Cell ◽

Lymphoid Tissue ◽

Ex Vivo ◽

Cd4 T Cell ◽

Cell Depletion ◽

T Cell Depletion ◽

Human Lymphoid Tissue ◽

Hiv 1

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Proline 35 of Human Immunodeficiency Virus Type 1 (HIV-1) Vpr Regulates the Integrity of the N-Terminal Helix and the Incorporation of Vpr into Virus Particles and Supports the Replication of R5-Tropic HIV-1 in Human Lymphoid Tissue Ex Vivo

Journal of Virology ◽

10.1128/jvi.02803-06 ◽

2007 ◽

Vol 81 (17) ◽

pp. 9572-9576 ◽

Cited By ~ 6

Author(s):

Jörg Votteler ◽

Nicole Studtrucker ◽

Stefan Sörgel ◽

Jan Münch ◽

Elke Rücker ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Lymphoid Tissue ◽

Mutational Analysis ◽

Ex Vivo ◽

Immunodeficiency Virus ◽

Human Lymphoid Tissue ◽

Hiv 1

ABSTRACT Mutational analysis of the four conserved proline residues in human immunodeficiency virus type 1 (HIV-1) Vpr reveals that only Pro-35 is required for efficient replication of R5-tropic, but not of X4-tropic, viruses in human lymphoid tissue (HLT) cultivated ex vivo. While Vpr-mediated apoptosis and G2 cell cycle arrest, as well as the expression and subcellular localization of Vpr, were independent, the capacity for encapsidation of Vpr into budding virions was dependent on Pro-35. 1H nuclear magnetic resonance data suggest that mutation of Pro-35 causes a conformational change in the hydrophobic core of the molecule, whose integrity is required for the encapsidation of Vpr, and thus, Pro-35 supports the replication of R5-tropic HIV-1 in HLT.

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R5 Variants of Human Immunodeficiency Virus Type 1 Preferentially Infect CD62L− CD4+ T Cells and Are Potentially Resistant to Nucleoside Reverse Transcriptase Inhibitors

Journal of Virology ◽

10.1128/jvi.80.2.854-865.2006 ◽

2006 ◽

Vol 80 (2) ◽

pp. 854-865 ◽

Cited By ~ 8

Author(s):

Françoise Gondois-Rey ◽

Angelique Biancotto ◽

Marcelo Antonio Fernandez ◽

Lise Bettendroffer ◽

Jana Blazkova ◽

...

Keyword(s):

T Cells ◽

Reverse Transcriptase ◽

Lymphoid Tissue ◽

Ex Vivo ◽

Productive Infection ◽

Reverse Transcriptase Inhibitors ◽

Hla Dr ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT The persistence of human immunodeficiency virus type 1 (HIV-1) in memory CD4+ T cells is a major obstacle to the eradication of the virus with current antiretroviral therapy. Here, we investigated the effect of the activation status of CD4+ T cells on the predominance of R5 and X4 HIV-1 variants in different subsets of CD4+ T cells in ex vivo-infected human lymphoid tissues and peripheral blood mononuclear cells (PBMCs). In these cell systems, we examined the sensitivity of HIV replication to reverse transcriptase inhibitors. We demonstrate that R5 HIV-1 variants preferentially produced productive infection in HLA-DR− CD62L− CD4+ T cells. These cells were mostly in the G1b phase of the cell cycle, divided slowly, and expressed high levels of CCR5. In contrast, X4 HIV-1 variants preferentially produced productive infection in activated HLA-DR+ CD62L+ CD4+ T cells, which expressed high levels of CXCR4. The abilities of the nucleoside reverse transcriptase inhibitors (NRTI) zidovudine and lamivudine to stop HIV-1 replication were 20 times greater in activated T cells than in slowly dividing HLA-DR− CD62L− CD4+ T cells. This result, demonstrated both in a highly physiologically relevant ex vivo lymphoid tissue model and in PBMCs, correlated with higher levels of thymidine kinase mRNA in activated than in slowly dividing HLA-DR− CD62L− CD4+ T cells. The non-NRTI nevirapine was equally efficient in both cell subsets. The lymphoid tissue and PBMC-derived cell systems represent well-defined models which could be used as new tools for the study of the mechanism of resistance to HIV-1 inhibitors in HLA-DR− CD62L− CD4+ T cells.

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Determinants in HIV-1 Nef for enhancement of virus replication and depletion of CD4+ T lymphocytes in human lymphoid tissue ex vivo

Retrovirology ◽

10.1186/1742-4690-6-6 ◽

2009 ◽

Vol 6 (1) ◽

pp. 6 ◽

Cited By ~ 21

Author(s):

Stefanie Homann ◽

Nadine Tibroni ◽

Ingo Baumann ◽

Serkan Sertel ◽

Oliver T Keppler ◽

...

Keyword(s):

T Lymphocytes ◽

Virus Replication ◽

Lymphoid Tissue ◽

Ex Vivo ◽

Human Lymphoid Tissue ◽

Hiv 1

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HIV-1–induced activation of CD4+ T cells creates new targets for HIV-1 infection in human lymphoid tissue ex vivo

Blood ◽

10.1182/blood-2007-05-088435 ◽

2008 ◽

Vol 111 (2) ◽

pp. 699-704 ◽

Cited By ~ 62

Author(s):

Angélique Biancotto ◽

Sarah J. Iglehart ◽

Christophe Vanpouille ◽

Cristian E. Condack ◽

Andrea Lisco ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Lymphoid Tissue ◽

Ex Vivo ◽

Lymphoid Tissues ◽

Strong Positive Correlation ◽

New Targets ◽

Hla Dr ◽

Human Lymphoid Tissue ◽

Hiv 1

We demonstrate mechanisms by which HIV-1 appears to facilitate its own infection in ex vivo–infected human lymphoid tissue. In this system, HIV-1 readily infects various CD4+ T cells, but productive viral infection was supported predominantly by activated T cells expressing either CD25 or HLA-DR or both (CD25/HLA-DR) but not other activation markers: There was a strong positive correlation (r = 0.64, P = .001) between virus production and the number of CD25+/HLA-DR+ T cells. HIV-1 infection of lymphoid tissue was associated with activation of both HIV-1–infected and uninfected (bystanders) T cells. In these tissues, apoptosis was selectively increased in T cells expressing CD25/HLA-DR and p24gag but not in cells expressing either of these markers alone. In the course of HIV-1 infection, there was a significant increase in the number of activated (CD25+/HLA-DR+) T cells both infected and uninfected (bystander). By inducing T cells to express particular markers of activation that create new targets for infection, HIV-1 generates in ex vivo lymphoid tissues a vicious destructive circle of activation and infection. In vivo, such self-perpetuating cycle could contribute to HIV-1 disease.

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Segregation of R5 and X4 HIV-1 variants to memory T cell subsets differentially expressing CD62L in ex vivo infected human lymphoid tissue

AIDS ◽

10.1097/00002030-200206140-00006 ◽

2002 ◽

Vol 16 (9) ◽

pp. 1245-1249 ◽

Cited By ~ 16

Author(s):

Françoise Gondois-Rey ◽

Jean-Charles Grivel ◽

Angelique Biancotto ◽

Marjorie Pion ◽

Robert Vigne ◽

...

Keyword(s):

T Cell ◽

Lymphoid Tissue ◽

Ex Vivo ◽

T Cell Subsets ◽

Memory T Cell ◽

Human Lymphoid Tissue ◽

Hiv 1

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Nef Enhances Human Immunodeficiency Virus Replication and Responsiveness to Interleukin-2 in Human Lymphoid Tissue Ex Vivo

Journal of Virology ◽

10.1128/jvi.73.5.3968-3974.1999 ◽

1999 ◽

Vol 73 (5) ◽

pp. 3968-3974 ◽

Cited By ~ 60

Author(s):

Svetlana Glushakova ◽

Jean-Charles Grivel ◽

Kalachar Suryanarayana ◽

Pascal Meylan ◽

Jeffrey D. Lifson ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

T Cells ◽

Lymphoid Tissue ◽

Ex Vivo ◽

Interleukin 2 ◽

Dependent Manner ◽

Immunodeficiency Virus ◽

Infected Cells ◽

Human Lymphoid Tissue ◽

Hiv 1

ABSTRACT The nef gene is important for the pathogenicity associated with simian immunodeficiency virus infection in rhesus monkeys and with human immunodeficiency virus type 1 (HIV-1) infection in humans. The mechanisms by which nef contributes to pathogenesis in vivo remain unclear. We investigated the contribution of nef to HIV-1 replication in human lymphoid tissue ex vivo by studying infection with parental HIV-1 strain NL4-3 and with anef mutant (ΔnefNL4-3). In human tonsillar histocultures, NL4-3 replicated to higher levels than ΔnefNL4-3 did. Increased virus production with NL4-3 infection was associated with increased numbers of productively infected cells and greater loss of CD4+ T cells over time. While the numbers of productively infected T cells were increased in the presence of nef, the levels of viral expression and production per infected T cell were similar whether the nefgene was present or not. Exogenous interleukin-2 (IL-2) increased HIV-1 production in NL4-3-infected tissue in a dose-dependent manner. In contrast, ΔnefNL4-3 production was enhanced only marginally by IL-2. Thus, Nef can facilitate HIV-1 replication in human lymphoid tissue ex vivo by increasing the numbers of productively infected cells and by increasing the responsiveness to IL-2 stimulation.

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