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Adipose Tissue Is a Neglected Viral Reservoir and an Inflammatory Site during Chronic HIV and SIV Infection.

Damouche A, Lazure T, Avettand-Fènoël V, Huot N, Dejucq-Rainsford N, Satie AP, Mélard A, David L, Gommet C, Ghosn J, Noel N, Pourcher G, Martinez V, Benoist S, Béréziat V, Cosma A, Favier B, Vaslin B, Rouzioux C, Capeau J, Müller-Trutwin M, Dereuddre-Bosquet N, Le Grand R, Lambotte O, Bourgeois C - PLoS Pathog. (2015)

Bottom Line: We detected cell-associated SIV DNA and RNA in the SVF and in sorted CD4+ T cells and macrophages from adipose tissue.Importantly, the production of HIV RNA was detected by in situ hybridization, and after the in vitro reactivation of sorted CD4+ T cells from adipose tissue.These observations open up new therapeutic strategies for limiting the size of the viral reservoir and decreasing low-grade chronic inflammation via the modulation of adipose tissue-related pathways.

View Article: PubMed Central - PubMed

Affiliation: Université Paris Sud, UMR 1184, Le Kremlin-Bicêtre, France; CEA, DSV/iMETI, IDMIT, Fontenay-aux-Roses, France; INSERM, U1184, Immunology of Viral Infections and Autoimmune Diseases, Le Kremlin-Bicêtre, France.

ABSTRACT
Two of the crucial aspects of human immunodeficiency virus (HIV) infection are (i) viral persistence in reservoirs (precluding viral eradication) and (ii) chronic inflammation (directly associated with all-cause morbidities in antiretroviral therapy (ART)-controlled HIV-infected patients). The objective of the present study was to assess the potential involvement of adipose tissue in these two aspects. Adipose tissue is composed of adipocytes and the stromal vascular fraction (SVF); the latter comprises immune cells such as CD4+ T cells and macrophages (both of which are important target cells for HIV). The inflammatory potential of adipose tissue has been extensively described in the context of obesity. During HIV infection, the inflammatory profile of adipose tissue has been revealed by the occurrence of lipodystrophies (primarily related to ART). Data on the impact of HIV on the SVF (especially in individuals not receiving ART) are scarce. We first analyzed the impact of simian immunodeficiency virus (SIV) infection on abdominal subcutaneous and visceral adipose tissues in SIVmac251 infected macaques and found that both adipocytes and adipose tissue immune cells were affected. The adipocyte density was elevated, and adipose tissue immune cells presented enhanced immune activation and/or inflammatory profiles. We detected cell-associated SIV DNA and RNA in the SVF and in sorted CD4+ T cells and macrophages from adipose tissue. We demonstrated that SVF cells (including CD4+ T cells) are infected in ART-controlled HIV-infected patients. Importantly, the production of HIV RNA was detected by in situ hybridization, and after the in vitro reactivation of sorted CD4+ T cells from adipose tissue. We thus identified adipose tissue as a crucial cofactor in both viral persistence and chronic immune activation/inflammation during HIV infection. These observations open up new therapeutic strategies for limiting the size of the viral reservoir and decreasing low-grade chronic inflammation via the modulation of adipose tissue-related pathways.

No MeSH data available.


Related in: MedlinePlus

HIV RNA production following in vitro reactivation.Total SVF (filled circles), sorted CD4+CD3+ (filled squares) and CD14+CD206+ cells (filled triangles) from adipose tissue, and sorted CD4+CD3+ (open squares) and CD14+ cells (open triangles) from PBMCs were co-cultured with allogeneic, pre-activated CD4+ T cells and then stimulated. The cell number differed according to the cell fraction: 1x106 for the total SVF fraction, 1x105 to 5x105 for sorted CD4+ T cells, and 5x104 to 2x105 for sorted, CD14-expressing cells. HIV RNA was detected in supernatants collected at D7, 11, 14, 17 and 21. As a negative control, 1x106 SVF cells were cultured in the absence of allogeneic, pre-activated CD4+ T cells (Ns SVF open circles). Samples from six ART-treated HIV infected patient were tested and the HIV RNA detection assay was performed five times for each sample.
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ppat.1005153.g008: HIV RNA production following in vitro reactivation.Total SVF (filled circles), sorted CD4+CD3+ (filled squares) and CD14+CD206+ cells (filled triangles) from adipose tissue, and sorted CD4+CD3+ (open squares) and CD14+ cells (open triangles) from PBMCs were co-cultured with allogeneic, pre-activated CD4+ T cells and then stimulated. The cell number differed according to the cell fraction: 1x106 for the total SVF fraction, 1x105 to 5x105 for sorted CD4+ T cells, and 5x104 to 2x105 for sorted, CD14-expressing cells. HIV RNA was detected in supernatants collected at D7, 11, 14, 17 and 21. As a negative control, 1x106 SVF cells were cultured in the absence of allogeneic, pre-activated CD4+ T cells (Ns SVF open circles). Samples from six ART-treated HIV infected patient were tested and the HIV RNA detection assay was performed five times for each sample.

Mentions: The ex vivo induction of viral replication in sorted cells is a further means of demonstrating that HIV is replication-competent. In six long-term ART-treated patients, we performed an in vitro viral reactivation assay on total SVF cells, sorted adipose CD4+ and CD206+CD14+ cells and sorted peripheral blood CD4+ or CD14+ cells in the presence of allogeneic pre-activated CD4+ T cells and phytohemagglutinin (Fig 8). In order to obtain sufficient numbers of cells, SCAT and VAT samples were pooled when both were available. Although small numbers of cells (5 104 to 5 105) were used for this reactivation assay, we detected the induction of viral replication in the sorted CD4+ T cell fraction from the SVF of all six patients. HIV RNA in supernatants from the ex vivo-activated, sorted SVF CD4+ T cells was detected from day 7 to day 21 in nearly all patients. In two patients (#5, 7), HIV RNA was detected solely after the reactivation of sorted SVF CD4+ T cells but not in total SVF or in PBMCs—probably because of the small number of infected cells in the culture. In the four other patients (#6, 8, 9, 10), HIV RNA was detected in similar numbers of ex vivo-activated CD4+ T cells recovered from both adipose tissue and PBMCs. Induction of HIV replication from SVF was detected in these four patients (albeit at low levels in three—probably due to the small number of infected cells). No HIV RNA was detected in cultures of activated adipose CD14+CD206+ fractions. Importantly, HIV RNA was not detected in non-activated SVF cell cultures (except in one patient), meaning that we could rule out the amplification of non-latent, pre-existing HIV. Our results show that adipose CD4+ T cells were infected by replication-competent HIV.


Adipose Tissue Is a Neglected Viral Reservoir and an Inflammatory Site during Chronic HIV and SIV Infection.

Damouche A, Lazure T, Avettand-Fènoël V, Huot N, Dejucq-Rainsford N, Satie AP, Mélard A, David L, Gommet C, Ghosn J, Noel N, Pourcher G, Martinez V, Benoist S, Béréziat V, Cosma A, Favier B, Vaslin B, Rouzioux C, Capeau J, Müller-Trutwin M, Dereuddre-Bosquet N, Le Grand R, Lambotte O, Bourgeois C - PLoS Pathog. (2015)

HIV RNA production following in vitro reactivation.Total SVF (filled circles), sorted CD4+CD3+ (filled squares) and CD14+CD206+ cells (filled triangles) from adipose tissue, and sorted CD4+CD3+ (open squares) and CD14+ cells (open triangles) from PBMCs were co-cultured with allogeneic, pre-activated CD4+ T cells and then stimulated. The cell number differed according to the cell fraction: 1x106 for the total SVF fraction, 1x105 to 5x105 for sorted CD4+ T cells, and 5x104 to 2x105 for sorted, CD14-expressing cells. HIV RNA was detected in supernatants collected at D7, 11, 14, 17 and 21. As a negative control, 1x106 SVF cells were cultured in the absence of allogeneic, pre-activated CD4+ T cells (Ns SVF open circles). Samples from six ART-treated HIV infected patient were tested and the HIV RNA detection assay was performed five times for each sample.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4581628&req=5

ppat.1005153.g008: HIV RNA production following in vitro reactivation.Total SVF (filled circles), sorted CD4+CD3+ (filled squares) and CD14+CD206+ cells (filled triangles) from adipose tissue, and sorted CD4+CD3+ (open squares) and CD14+ cells (open triangles) from PBMCs were co-cultured with allogeneic, pre-activated CD4+ T cells and then stimulated. The cell number differed according to the cell fraction: 1x106 for the total SVF fraction, 1x105 to 5x105 for sorted CD4+ T cells, and 5x104 to 2x105 for sorted, CD14-expressing cells. HIV RNA was detected in supernatants collected at D7, 11, 14, 17 and 21. As a negative control, 1x106 SVF cells were cultured in the absence of allogeneic, pre-activated CD4+ T cells (Ns SVF open circles). Samples from six ART-treated HIV infected patient were tested and the HIV RNA detection assay was performed five times for each sample.
Mentions: The ex vivo induction of viral replication in sorted cells is a further means of demonstrating that HIV is replication-competent. In six long-term ART-treated patients, we performed an in vitro viral reactivation assay on total SVF cells, sorted adipose CD4+ and CD206+CD14+ cells and sorted peripheral blood CD4+ or CD14+ cells in the presence of allogeneic pre-activated CD4+ T cells and phytohemagglutinin (Fig 8). In order to obtain sufficient numbers of cells, SCAT and VAT samples were pooled when both were available. Although small numbers of cells (5 104 to 5 105) were used for this reactivation assay, we detected the induction of viral replication in the sorted CD4+ T cell fraction from the SVF of all six patients. HIV RNA in supernatants from the ex vivo-activated, sorted SVF CD4+ T cells was detected from day 7 to day 21 in nearly all patients. In two patients (#5, 7), HIV RNA was detected solely after the reactivation of sorted SVF CD4+ T cells but not in total SVF or in PBMCs—probably because of the small number of infected cells in the culture. In the four other patients (#6, 8, 9, 10), HIV RNA was detected in similar numbers of ex vivo-activated CD4+ T cells recovered from both adipose tissue and PBMCs. Induction of HIV replication from SVF was detected in these four patients (albeit at low levels in three—probably due to the small number of infected cells). No HIV RNA was detected in cultures of activated adipose CD14+CD206+ fractions. Importantly, HIV RNA was not detected in non-activated SVF cell cultures (except in one patient), meaning that we could rule out the amplification of non-latent, pre-existing HIV. Our results show that adipose CD4+ T cells were infected by replication-competent HIV.

Bottom Line: We detected cell-associated SIV DNA and RNA in the SVF and in sorted CD4+ T cells and macrophages from adipose tissue.Importantly, the production of HIV RNA was detected by in situ hybridization, and after the in vitro reactivation of sorted CD4+ T cells from adipose tissue.These observations open up new therapeutic strategies for limiting the size of the viral reservoir and decreasing low-grade chronic inflammation via the modulation of adipose tissue-related pathways.

View Article: PubMed Central - PubMed

Affiliation: Université Paris Sud, UMR 1184, Le Kremlin-Bicêtre, France; CEA, DSV/iMETI, IDMIT, Fontenay-aux-Roses, France; INSERM, U1184, Immunology of Viral Infections and Autoimmune Diseases, Le Kremlin-Bicêtre, France.

ABSTRACT
Two of the crucial aspects of human immunodeficiency virus (HIV) infection are (i) viral persistence in reservoirs (precluding viral eradication) and (ii) chronic inflammation (directly associated with all-cause morbidities in antiretroviral therapy (ART)-controlled HIV-infected patients). The objective of the present study was to assess the potential involvement of adipose tissue in these two aspects. Adipose tissue is composed of adipocytes and the stromal vascular fraction (SVF); the latter comprises immune cells such as CD4+ T cells and macrophages (both of which are important target cells for HIV). The inflammatory potential of adipose tissue has been extensively described in the context of obesity. During HIV infection, the inflammatory profile of adipose tissue has been revealed by the occurrence of lipodystrophies (primarily related to ART). Data on the impact of HIV on the SVF (especially in individuals not receiving ART) are scarce. We first analyzed the impact of simian immunodeficiency virus (SIV) infection on abdominal subcutaneous and visceral adipose tissues in SIVmac251 infected macaques and found that both adipocytes and adipose tissue immune cells were affected. The adipocyte density was elevated, and adipose tissue immune cells presented enhanced immune activation and/or inflammatory profiles. We detected cell-associated SIV DNA and RNA in the SVF and in sorted CD4+ T cells and macrophages from adipose tissue. We demonstrated that SVF cells (including CD4+ T cells) are infected in ART-controlled HIV-infected patients. Importantly, the production of HIV RNA was detected by in situ hybridization, and after the in vitro reactivation of sorted CD4+ T cells from adipose tissue. We thus identified adipose tissue as a crucial cofactor in both viral persistence and chronic immune activation/inflammation during HIV infection. These observations open up new therapeutic strategies for limiting the size of the viral reservoir and decreasing low-grade chronic inflammation via the modulation of adipose tissue-related pathways.

No MeSH data available.


Related in: MedlinePlus