The Genome-wide Methylation Profile of CD4+ T Cells From Individuals With Human Immunodeficiency Virus (HIV) Identifies Distinct Patterns Associated With Disease Progression

Background: Human genetic variation-mostly in the HLA and CCR5 regions-explains 25% of the variability in progression of HIV infection. However, it is also known that viral infections can modify cellular DNA methylation patterns. Therefore, changes in the methylation of CpG islands might modulate progression of HIV infection. Methods: 85 samples were analyzed: 21 elite controllers (EC), 21 HIV-infected subjects before combination antiretroviral therapy (cART) (viremic, 93,325 HIV-1 RNA copies/ml) and under suppressive cART (cART, median of 17 months, <50 HIV-1 RNA copies/ml), and 22 HIV-negative donors (HIVneg). We analyzed the methylation pattern of 485,577 CpG in DNA from peripheral CD4+ T lymphocytes. We selected the most differentially methylated gene (TNF) and analyzed its specific methylation, mRNA expression, and plasma protein levels in 5 individuals before and after initiation of cART. Results: We observed 129 methylated CpG sites (associated with 43 gene promoters) for which statistically significant differences were recorded in viremic vs HIVneg, 162 CpG sites (55 gene promoters) in viremic vs cART, 441 CpG sites (163 gene promoters) in viremic vs EC, but none in EC vs HIVneg. The TNF promoter region was hypermethylated in viremic vs HIVneg, cART, and EC. Moreover, we observed greater plasma levels of TNF in viremic individuals than in EC, cART, and HIVneg. Conclusions: Our study shows that genome methylation patterns vary depending on HIV infection status and progression profile and that these variations might have an impact on controlling HIV infection in the absence of cART

Estudos sobre aspectos virais e genéticos relacionados à integrase e ao processo de integração do HIV-1

Made available in DSpace on 2015-11-11T12:13:56Z (GMT). No. of bitstreams: 2 caroline_passaes_ioc_dout_2012.pdf: 6197462 bytes, checksum: 60b931e8b71d31d134ea1576e4b43737 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2015-05-21Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.A integrase (IN) é uma enzima chave para o ciclo de replicação do HIV, sendo responsável por catalisar a integração do genoma do HIV no cromossomo hospedeiro. Devido ao papel essencial desta enzima para a patogênese da infecção pelo HIV, a recente introdução dos inibidores de IN (INI) na prática clínica e em vista da escassez de informação sobre a diversidade genética da IN do HIV no Brasil, o presente estudo tem como objetivos a) investigar a diversidade genética da IN e os níveis de resistência primária nos subtipos B, C e F do HIV que são prevalentes no Brasil; b) acompanhar pacientes sob terapia antirretroviral em esquemas contendo raltegravir (RAL) a fim de monitorar a emergência de mutações de resistência aos INI; c) desenvolver um método de genotipagem da IN do HIV para ser usado na pratica clínica no Brasil; e d) investigar o envolvimento do processo de integração no controle da replicação do HIV. Não foram detectadas mutações principais associadas aos INI entre os indivíduos virgens de tratamento infectados com diferentes subtipos de HIV-1 O nível de mutações acessórias observadas foi bem baixo, e algumas posições foram polimórficas nas amostras brasileiras dos subtipos B, C e F. Esses resultados encorajam o uso de INI no Brasil. Analisando as coortes de pacientes que trocaram a enfuvirtida por RAL ou sob terapia de resgate com RAL, nós observamos um aumento nas contagens de células T CD4+ e uma rápida diminuição da carga viral no grupo sob terapia de resgate. Três pacientes não atingiram supressão virológica e as mutações Q148H+G140S foram detectadas em dois deles. A fim de monitorar o crescente número de pacientes sob terapia com RAL no Brasil, nós desenvolvemos um método de genotipagem in-house que está atualmente em teste pela rede de Genotipagem do Ministério da Saúde do Brasil para futura incorporação no monitoramento de pacientes falhando INI. Sobre o papel da IN no controle da infecção pelo HIV, não observamos mutações nos resíduos importantes para a atividade catalítica nas sequências de IN obtidas de pacientes controladores da infecção pelo HIV, nem acúmulo de DNA 2-LTR, sugerindo que não há um mecanismo bloqueando a integração nestes pacientes. Juntos, os resultados apresentados trazem informações importantes sobre a diversidade genética da IN, resistência aos INI e sobre o papel da IN na patogênese da infecção pelo HIVI ntegrase ( IN) is a key enzyme for the HIV - 1 replication cycle, being responsible for catalyz ing the integration of HIV genome into the host chromat in. Due to the essential role of this enzyme for HIV pathogenesis , the recent introduction of IN inhibitors in the clinical practice and in view of the paucity of information about the genetic diversity of HIV IN in Brazil, the present study aims to a) inv estigate the genetic diversity of IN and the levels of primary resistance in HIV subtypes B, C and F that are prevalent in Brazil; b) follow - up patients under antirretroviral therapy schemes containing raltegravir (RAL) in order to monitor the emergence of resistance mutations to IN inhibitor (INI) ; c) develop a method to genotype HIV IN to be used in c linical practice in Brazil; and d) investigate the involvement of the integration process in the control of HIV replication. No major resistance mutations as sociated to IN I w ere detected among drug - naïve individuals infected with distinct HIV - 1 subtypes . The level of accessory mutations was very low, and some positions were polymorphic in Brazilian samples of HIV subtypes B, C and F. These results encourage th e use of IN I in Brazil. By a nalyzing cohorts of patients that switched from enfuvirtide to RAL or under salvage therapy with RAL we observed an increase in T CD4+ cell counts and a rapid decay of viral load in this salvage therapy group. For three patients , virological suppression was not achieved and the mutations Q148H+G140S were detected for two of them . In order to monitor the increasing number of patients under therapy with RAL in Brazil, we developed a n in - house genotyping method that is currently und er test by the Brazilian Ministry of Health Genotyping N etwork for further incorporation for the management of HIV patients failing INI . Concerning the role of the IN in the control of HIV infection, we did not observe mutations at residues important for c atalytic activity in the integrase sequences obtaine d from HIV controller patients or an accumulation of 2 - LTR DNA, suggesting that there is not a mechanism blocking HIV integration in these patients. Taken together, the results here presented bring import ant insights about HIV IN diversity, resistance to INI and the role of IN in HIV pathogenesi

HIV cure research: advances and prospects.

International audienceThirty years after the identification of HIV, a cure for HIV infection is still to be achieved. Advances of combined antiretroviral therapy (cART) in recent years have transformed HIV infection into a chronic disease when treatment is available. However, in spite of the favorable outcomes provided by the newer therapies, cART is not curative and patients are at risk of developing HIV-associated disorders. Moreover, universal access to antiretroviral treatment is restricted by financial obstacles. This review discusses the most recent strategies that have been developed in the search for an HIV cure and to improve life quality of people living with HIV

COVID-19 Research: Lessons from Non-Human Primate Models

International audienceSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the coronavirus disease 2019 (COVID-19). It emerged from China in December 2019 and rapidly spread across the globe, causing a pandemic with unprecedented impacts on public health and economy. Therefore, there is an urgent need for the development of curative treatments and vaccines. In humans, COVID-19 pathogenesis shows a wide range of symptoms, from asymptomatic to severe pneumonia. Identifying animal models of SARS-CoV-2 infection that reflect the clinical symptoms of COVID-19 is of critical importance. Nonhuman primates (NHPss) correspond to relevant models to assess vaccine and antiviral effectiveness. This review discusses the use of NHPs as models for COVID-19 research, with focus on the pathogenesis of SARS-CoV-2 infection, drug discovery and pre-clinical evaluation of vaccine candidates

NK cell spatial dynamics and IgA responses in gut-associated lymphoid tissues during SIV infections

International audienceAbstract HIV infection induces tissue damage including lymph node (LN) fibrosis and intestinal epithelial barrier disruption leading to bacterial translocation and systemic inflammation. Natural hosts of SIV, such as African Green Monkeys (AGM), do not display tissue damage despite high viral load in blood and intestinal mucosa. AGM mount a NK cell-mediated control of SIVagm replication in peripheral LN. We analyzed if NK cells also control SIVagm in mesenteric (mes) LN and if this has an impact on gut humoral responses and the production of IgA known for their anti-inflammatory role in the gut. We show that CXCR5 + NK cell frequencies increase in mesLN upon SIVagm infection and that NK cells migrate into and control viral replication in B cell follicles (BCF) of mesLN. The proportion of IgA+ memory B cells were increased in mesLN during SIVagm infection in contrast to SIVmac infection. Total IgA levels in gut remained normal during SIVagm infection, while strongly decreased in intestine of chronically SIVmac-infected macaques. Our data suggest an indirect impact of NK cell-mediated viral control in mesLN during SIVagm infection on preserved BCF function and IgA production in intestinal tissues

Temporal dynamics of HIV-1 circulating subtypes in distinct exposure categories in southern Brazil

Abstract Background The HIV-1 epidemic in Brazil is predominantly driven by subtype B. However, in Brazilian Southern region subtype C prevails and a relatively high AIDS incidence rate is observed. The aim of the present study was to assess the temporal dynamics of HIV-1 subtypes circulating in patients from distinct exposure categories in Southern Brazil. For this purpose 166 HIV-1 samples collected at the years of 1998 (group I) and 2005–2008 (group II) were analyzed. Results Analysis of group I revealed statistically significant (p Conclusions The present study shows an association between HIV subtypes and exposure categories at the middle 1990s in Southern Brazil. Our findings suggest that MSM and IDU populations might have played a major role in the introduction and initial dissemination of subtypes B and C, respectively, in Southern Brazil. This study also suggests a trend towards homogenization of HIV-1 strains across distinct exposure categories as a consequence of an overall increase in the prevalence of subtype C and BC recombinants in both HET and MSM populations.</p

Anti-apoptotic clone 11 derived peptides induce in vitro death of CD4+ T cells susceptible to HIV-1 infection

International audienceHIV-1 successfully establishes long-term infection in its target cells despite viral cytotoxic effects. We have recently shown that cell metabolism is an important factor driving CD4+ T-cell susceptibility to HIV-1 and the survival of infected cells. We show here that expression of anti-apoptotic clone 11 (AAC-11), an anti-apoptotic factor upregulated in many cancers, increased with progressive CD4+ T cell memory differentiation in association with the expression of cell cycle, activation and metabolism genes and correlated with susceptibility to HIV-1 infection. Synthetic peptides based on the LZ domain sequence of AAC-11, responsible for its interaction with molecular partners, were previously shown to be cytotoxic to cancer cells. Here we observed that these peptides also blocked HIV-1 infection by inducing cell death of HIV-1 susceptible primary CD4+ T-cells across all T-cell subsets. The peptides targeted metabolically active cells and had the greatest effect on effector and transitional CD4+ T cell memory subsets. Our results suggest that AAC-11 survival pathway is potentially involved in the survival of HIV-1 infectable cells and provide a proof of principle that some cellular characteristics can be targeted to eliminate the cells offering the best conditions to sustain HIV-1 replication.IMPORTANCE Although antiretroviral treatment efficiently blocks HIV multiplication, it cannot eliminate the cells already carrying integrated proviruses. In the search for a HIV cure the identification of new potential targets to selectively eliminate infected cells is of the outmost importance. We show here that peptides derived from the anti-apoptotic clone 11 (AAC-11), which expression levels correlated with susceptibility to HIV-1 infection of CD4+ T-cells, induced cytotoxicity in CD4+ T-cells showing the highest levels of activation and metabolic activity, conditions known to favor HIV-1 infection. Accordingly, CD4+ T-cells that survived the cytotoxic action of the AAC-11 peptides were resistant to HIV-1 replication. Our results identify a new potential molecular pathway to target HIV-1 infection

Temporal dynamics of HIV-1 circulating subtypes in distinct exposure categories in southern Brazil

Submitted by Sandra Infurna ([email protected]) on 2016-09-15T15:14:30Z No. of bitstreams: 1 caroline2_passaes_etal_IOC_2012.pdf: 375073 bytes, checksum: 0a8be9d439c43d6fb830774c41d06c6a (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2016-09-15T15:26:39Z (GMT) No. of bitstreams: 1 caroline2_passaes_etal_IOC_2012.pdf: 375073 bytes, checksum: 0a8be9d439c43d6fb830774c41d06c6a (MD5)Made available in DSpace on 2016-09-15T15:26:39Z (GMT). No. of bitstreams: 1 caroline2_passaes_etal_IOC_2012.pdf: 375073 bytes, checksum: 0a8be9d439c43d6fb830774c41d06c6a (MD5) Previous issue date: 2012Fundação Estadual de Produção e Pesquisa em Saúde – FEPPS. Centro de Desenvolvimento Técnico e Científico – CDCT. Porto Alegre, RS, Brasil.Fundação Estadual de Produção e Pesquisa em Saúde – FEPPS. Centro de Desenvolvimento Técnico e Científico – CDCT. Porto Alegre, RS, Brasil / Universidade Federal do Rio Grande do Sul (UFRGS). Centro de Ciências Biológicas. Departamento de Genética. Programa de Pós-Graduação em Genética e Biologia Molecular. Porto Alegre, RS, Brasil.Fundação Estadual de Produção e Pesquisa em Saúde – FEPPS. Centro de Desenvolvimento Técnico e Científico – CDCT. Porto Alegre, RS, Brasil / Universidade Federal do Rio Grande do Sul (UFRGS). Centro de Ciências Biológicas. Departamento de Genética. Programa de Pós-Graduação em Genética e Biologia Molecular. Porto Alegre, RS, Brasil.Fundação Estadual de Produção e Pesquisa em Saúde – FEPPS. Centro de Desenvolvimento Técnico e Científico – CDCT. Porto Alegre, RS, Brasil / Universidade Federal de Santa Catarina (UFSC). Centro de Ciências Biológicas. Departamento de Microbiologia, Imunologia e Parasitologia. Programa de Pós-Graduação em Biotecnologia e Biociências. Florianópolis, SC, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil

TLR7 dosage polymorphism shapes interferogenesis and HIV-1 acute viremia in women

International audienceType I IFN (IFN-I) production by plasmacytoid DCs (pDCs) occurs during acute HIV-1 infection in response to TLR7 stimulation, but the role of pDC-derived IFN-I in controlling or promoting HIV-1 infection is ambiguous. We report here a sex-biased interferogenic phenotype for a frequent single-nucleotide polymorphism of human TLR7, rs179008, displaying an impact on key parameters of acute HIV-1 infection. We show allele rs179008 T to determine lower TLR7 protein abundance in cells from women, specifically — likely by diminishing TLR7 mRNA translation efficiency through codon usage. The hypomorphic TLR7 phenotype is mirrored by decreased TLR7-driven IFN-I production by female pDCs. Among women from the French ANRS PRIMO cohort of acute HIV-1 patients, carriage of allele rs179008 T associated with lower viremia, cell-associated HIV-1 DNA, and CXCL10 (IP-10) plasma concentrations. RNA viral load was decreased by 0.85 log10 (95% CI, −1.51 to −0.18) among T/T homozygotes, who also exhibited a lower frequency of acute symptoms. TLR7 emerges as an important control locus for acute HIV-1 viremia, and the clinical phenotype for allele rs179008 T, carried by 30%–50% of European women, supports a beneficial effect of toning down TLR7-driven IFN-I production by pDCs during acute HIV-1 infection

HIV controllers with different viral load cut-off levels have distinct virologic and immunologic profiles

Submitted by sandra infurna ([email protected]) on 2016-04-05T13:12:25Z No. of bitstreams: 1 fernanda_cortes_etal_IOC_2015.pdf: 720405 bytes, checksum: e4ef65f6aa4098f1ff80207771af65c0 (MD5)Approved for entry into archive by sandra infurna ([email protected]) on 2016-04-05T13:31:20Z (GMT) No. of bitstreams: 1 fernanda_cortes_etal_IOC_2015.pdf: 720405 bytes, checksum: e4ef65f6aa4098f1ff80207771af65c0 (MD5)Made available in DSpace on 2016-04-05T13:31:20Z (GMT). No. of bitstreams: 1 fernanda_cortes_etal_IOC_2015.pdf: 720405 bytes, checksum: e4ef65f6aa4098f1ff80207771af65c0 (MD5) Previous issue date: 2015Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas - INI. Rio de Janeiro, RJ, Brasil.University of Miami. Miller School of Medicine. Department of Medicine. Miami, FL, USA.University of Miami. Miller School of Medicine. Department of Medicine. Miami, FL, USA.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas - INI. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas - INI. Rio de Janeiro, RJ, Brasil.University of Miami. Miller School of Medicine. Department of Medicine. Miami, FL, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Background—The mechanisms behind natural control of HIV replication are still unclear, and several studies pointed that elite controllers are a heterogeneous group. Methods—We performed analyses of virologic, genetic and immunologic parameters of HIV-1 controllers groups: 1) Elite Controllers (EC; VL <80 copies/mL); 2) Ebbing Elite Controllers (EEC; transient viremia/blips); and Viremic Controllers (VC; detectable viremia <5,000 copies/ mL). Untreated non-controllers (NC), patients under suppressive HAART and HIV-1 negative individuals were analyzed as controls. Results—Total and integrated HIV-1 DNA for EC were significantly lower than for NC and HAART groups. 2-LTR circles were detected in EEC (3/5) and VC (6/7) but not in EC. While EC and EEC maintain normal T cell counts over time, some VC displayed negative CD4+ T cells slopes. VC and EEC showed a higher percentage of activated CD8+ T cells and microbial translocation than HIV-1 negative controls. EC displayed a weaker Gag/Nef IFN-γ T cell response and a significantly lower proportion of anti-HIV IgG antibodies than EEC, VC and NC groups. Conclusion—Transient/persistent low level viremia in HIV controllers may have an impact on immunologic and virologic profiles. Classify HIV controllers patients taking into account their virologic profile may decrease the heterogeneity of HIV controllers cohorts, which may help to clarify the mechanisms associated to the elite control of HIV