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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

Detection of HIV DNA and RNA in adipose tissue.(A) Detection of HIV DNA in SVF cells and PBMCs from 11 ART-treated, HIV-infected patients. Each patient is represented by a symbol, and the shading represents the type of adipose tissue: SCAT (open symbols), VAT (filled symbols). (B) HIV DNA detection in sorted CD4+ T cells recovered from adipose tissue and PBMCs from 3 ART-treated, HIV-infected patients. The HIV DNA detection assay was performed in duplicate and is expressed in log copies per million cells. The detection limit differed as a function of the numbers of cells tested and is indicated as <dl on the graph. (C) An in situ hybridization assay for HIV RNA was performed on three samples of adipose tissue recovered from ART-treated, HIV-infected patients (1 VAT and 2 SCAT samples). Positive control: a prostate tissue sample from a viremic patient. One slide was analyzed for each sample. HIV RNA staining for each positive patient is shown. Scale bar: 50 μm.
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ppat.1005153.g007: Detection of HIV DNA and RNA in adipose tissue.(A) Detection of HIV DNA in SVF cells and PBMCs from 11 ART-treated, HIV-infected patients. Each patient is represented by a symbol, and the shading represents the type of adipose tissue: SCAT (open symbols), VAT (filled symbols). (B) HIV DNA detection in sorted CD4+ T cells recovered from adipose tissue and PBMCs from 3 ART-treated, HIV-infected patients. The HIV DNA detection assay was performed in duplicate and is expressed in log copies per million cells. The detection limit differed as a function of the numbers of cells tested and is indicated as <dl on the graph. (C) An in situ hybridization assay for HIV RNA was performed on three samples of adipose tissue recovered from ART-treated, HIV-infected patients (1 VAT and 2 SCAT samples). Positive control: a prostate tissue sample from a viremic patient. One slide was analyzed for each sample. HIV RNA staining for each positive patient is shown. Scale bar: 50 μm.

Mentions: To ascertain whether adipose tissue may constitute a viral reservoir, we collected adipose tissue samples from 13 ART-treated HIV-infected patients who had undergone elective visceral surgery for non-HIV related causes (Table 1). In 11 patients, we searched for viral DNA in SVF cells and PBMCs. Even after clinically effective ART treatment, HIV DNA was detected in the SVF of all samples tested (11 from VAT and 4 from SCAT) (Fig 7A). The median level of HIV DNA in SVF from VAT (2.15 [2.01–2.47] log DNA copies/million cells) was significantly lower than in PBMCs (2.94 [2.25–3.28]). However, this low level of infection in adipose tissue might primarily reflect the low proportion of hematopoietic cells in adipose tissue. We therefore quantified HIV DNA in sorted CD4+ T cells and sorted CD206+ CD14+ cells from three patients. Contamination by circulating monocytes was controlled for by measuring CD206 expression on CD14+ cells. In one sample, HIV DNA could not be detected in adipose CD4+ T cells—presumably because of low DNA input. In the samples with detectable virus, the median level of HIV DNA in CD4+ T cell fractions recovered from adipose tissue was 3.83 log DNA copies/million cells (compared with 3.56 in sorted peripheral blood CD4+ T cells). We failed to detect HIV DNA in the three sorted CD14+ CD206+ samples tested. Therefore, in patients on long-term ART with no detectable viremia, we confirmed that HIV-infected cells are present in the SVF (mainly tissue-resident CD4+ T cells). Given that the detected HIV DNA might have been replication-defective, we also screened for viral RNA in adipose tissue sections using in situ hybridization. Two SCAT samples and one VAT sample were obtained from ART-treated HIV-infected patients. As shown in Fig 7C, cells expressing HIV RNA were detected (albeit in low numbers) in the three adipose tissue samples studied. We therefore demonstrated that productive, HIV-infected cells are present in the adipose tissue of ART-treated patients.


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)

Detection of HIV DNA and RNA in adipose tissue.(A) Detection of HIV DNA in SVF cells and PBMCs from 11 ART-treated, HIV-infected patients. Each patient is represented by a symbol, and the shading represents the type of adipose tissue: SCAT (open symbols), VAT (filled symbols). (B) HIV DNA detection in sorted CD4+ T cells recovered from adipose tissue and PBMCs from 3 ART-treated, HIV-infected patients. The HIV DNA detection assay was performed in duplicate and is expressed in log copies per million cells. The detection limit differed as a function of the numbers of cells tested and is indicated as <dl on the graph. (C) An in situ hybridization assay for HIV RNA was performed on three samples of adipose tissue recovered from ART-treated, HIV-infected patients (1 VAT and 2 SCAT samples). Positive control: a prostate tissue sample from a viremic patient. One slide was analyzed for each sample. HIV RNA staining for each positive patient is shown. Scale bar: 50 μm.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4581628&req=5

ppat.1005153.g007: Detection of HIV DNA and RNA in adipose tissue.(A) Detection of HIV DNA in SVF cells and PBMCs from 11 ART-treated, HIV-infected patients. Each patient is represented by a symbol, and the shading represents the type of adipose tissue: SCAT (open symbols), VAT (filled symbols). (B) HIV DNA detection in sorted CD4+ T cells recovered from adipose tissue and PBMCs from 3 ART-treated, HIV-infected patients. The HIV DNA detection assay was performed in duplicate and is expressed in log copies per million cells. The detection limit differed as a function of the numbers of cells tested and is indicated as <dl on the graph. (C) An in situ hybridization assay for HIV RNA was performed on three samples of adipose tissue recovered from ART-treated, HIV-infected patients (1 VAT and 2 SCAT samples). Positive control: a prostate tissue sample from a viremic patient. One slide was analyzed for each sample. HIV RNA staining for each positive patient is shown. Scale bar: 50 μm.
Mentions: To ascertain whether adipose tissue may constitute a viral reservoir, we collected adipose tissue samples from 13 ART-treated HIV-infected patients who had undergone elective visceral surgery for non-HIV related causes (Table 1). In 11 patients, we searched for viral DNA in SVF cells and PBMCs. Even after clinically effective ART treatment, HIV DNA was detected in the SVF of all samples tested (11 from VAT and 4 from SCAT) (Fig 7A). The median level of HIV DNA in SVF from VAT (2.15 [2.01–2.47] log DNA copies/million cells) was significantly lower than in PBMCs (2.94 [2.25–3.28]). However, this low level of infection in adipose tissue might primarily reflect the low proportion of hematopoietic cells in adipose tissue. We therefore quantified HIV DNA in sorted CD4+ T cells and sorted CD206+ CD14+ cells from three patients. Contamination by circulating monocytes was controlled for by measuring CD206 expression on CD14+ cells. In one sample, HIV DNA could not be detected in adipose CD4+ T cells—presumably because of low DNA input. In the samples with detectable virus, the median level of HIV DNA in CD4+ T cell fractions recovered from adipose tissue was 3.83 log DNA copies/million cells (compared with 3.56 in sorted peripheral blood CD4+ T cells). We failed to detect HIV DNA in the three sorted CD14+ CD206+ samples tested. Therefore, in patients on long-term ART with no detectable viremia, we confirmed that HIV-infected cells are present in the SVF (mainly tissue-resident CD4+ T cells). Given that the detected HIV DNA might have been replication-defective, we also screened for viral RNA in adipose tissue sections using in situ hybridization. Two SCAT samples and one VAT sample were obtained from ART-treated HIV-infected patients. As shown in Fig 7C, cells expressing HIV RNA were detected (albeit in low numbers) in the three adipose tissue samples studied. We therefore demonstrated that productive, HIV-infected cells are present in the adipose tissue of ART-treated patients.

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