PLoS Pathog

The low pathogenicity and replicative potential of HIV-2 are still poorly understood. We investigated whether HIV-2 reservoirs might follow the peculiar distribution reported in models of attenuated HIV-1/SIV infections, i.e. limited infection of central-memory CD4 T lymphocytes (TCM). Antiretroviral-naive HIV-2 infected individuals from the ANRS-CO5 (12 non-progressors, 2 progressors) were prospectively included. Peripheral blood mononuclear cells (PBMCs) were sorted into monocytes and resting CD4 T-cell subsets (naive [TN], central- [TCM], transitional- [TTM] and effector-memory [TEM]). Reactivation of HIV-2 was tested in 30-day cultures of CD8-depleted PBMCs. HIV-2 DNA was quantified by real-time PCR. Cell surface markers, co-receptors and restriction factors were analyzed by flow-cytometry and multiplex transcriptomic study. HIV-2 DNA was undetectable in monocytes from all individuals and was quantifiable in TTM from 4 individuals (median: 2.25 log10 copies/106 cells [IQR: 1.99-2.94]) but in TCM from only 1 individual (1.75 log10 copies/106 cells). HIV-2 DNA levels in PBMCs (median: 1.94 log10 copies/106 PBMC [IQR = 1.53-2.13]) positively correlated with those in TTM (r = 0.66, p = 0.01) but not TCM. HIV-2 reactivation was observed in the cells from only 3 individuals. The CCR5 co-receptor was distributed similarly in cell populations from individuals and donors. TCM had a lower expression of CXCR6 transcripts (p = 0.002) than TTM confirmed by FACS analysis, and a higher expression of TRIM5 transcripts (p = 0.004). Thus the low HIV-2 reservoirs differ from HIV-1 reservoirs by the lack of monocytic infection and a limited infection of TCM associated to a lower expression of a potential alternative HIV-2 co-receptor, CXCR6 and a higher expression of a restriction factor, TRIM5. These findings shed new light on the low pathogenicity of HIV-2 infection suggesting mechanisms close to those reported in other models of attenuated HIV/SIV infection models

Caractérisation immunologique des réservoirs des virus de l’immunodéficience humaine VIH-2 et VIH-1 dans les cellules immunocompétentes et étude des paramètres de l’immunité innée et intrinsèque

HIV-2 and HIV-1 differ in pathogenicity. To better understand the mechanisms of HIV- 2 low pathogenicity and replication potential, we studied immune and virologic characteristics of HIV-2 reservoirs of infected patients (cohort ANRS CO-05). Fourteen HIV-2+ subjects from the ANRS CO-05 cohort were compared to healthy donors. Seven peripheral blood sub-populations were sorted by flow cytometry : Monocytes, activated and resting CD4 T cells (TN, TCM, TTM, TEM). Viral DNA was quantified in sorted cells by PCR. HIV-2 DNA was detected in TTM and TCM sub-populations of 4 patients but not in monocytes. The phenotype of peripheral NK cells was analyzed by flow cytometry (24 HIV-2 ANRS CO-05 cohort). HIV-2+ patients had lower proportions of NKp30+ NK cells, increased production of IFN-g and altered cytolytic function in the presence B7-H6 surface ligand, when compared to HIV-1+. The analysis of HIV-1 and HIV-2 co-receptors and transcription factors expression showed higher values in HIV-1LTNP (ANRS CO15) cohort, compared to HIV-2+ and healthy donors. We also found that the PD1 expression by CD4 T cells was higher in HIV-1LTNP subjects compared to healthy donors, though not correlated to the HIV-1 plasma load. Her we show that HIV-2 reservoirs are distinct from the HIV-1 reservoirs by the absence of monocytic infection and a limited TCM infection.Les VIH-2 et VIH-1 diffèrent par leur pathogénicité. Pour mieux comprendre les mécanismes de faible pathogénicité et de réplication du VIH-2, nous avons étudié les caractéristiques immuno-virologiques des réservoirs du VIH-2 chez des patients VIH-2+ (ANRS-CO-05). Quatorze sujets VIH-2+ ont été comparés à des donneurs sains. Les cellules sanguines ont été triées en sept sous-populations (monocytes, T CD4 activés et au repos (TN, TCM, TTM, TEM)). L’ADN du VIH-2 a été quantifié par PCR et détecté chez 4 sujets : dans les TTM et les TCM mais pas dans les monocytes. L’analyse phénotypique des cellules NK a été réalisée par cytométrie en flux (24 sujets, ANRS-CO-05). Les cellules NK des VIH-2+ avaient : une expression de NKp30 diminuée, une augmentation de la production d'IFN-g et une altération de la fonction cytolytique en présence de B7-H6, comparés aux VIH-1+. L’expression des corécepteurs et des facteurs de transcription a été analysée, montrant des niveaux plus élevés chez les VIH-1 LTNP comparée aux VIH-2+, et aux donneurs sains pour la plupart des paramètres. Nous avons observé que l’expression de PD1 était plus importante sur les T CD4 des VIH-1 LTNP comparés aux donneurs sains. Nous avons montré que les faibles réservoirs du VIH-2 se distinguent des réservoirs du VIH-1 par l'absence d'infection monocytaire et par une infection limitée des TCM

Immunological characterization of HIV-2 and HIV-1 human immunodeficiency virus reservoirs in immunocompetent cells and study of innate and intrinsic immunity parameters

Les VIH-2 et VIH-1 diffèrent par leur pathogénicité. Pour mieux comprendre les mécanismes de faible pathogénicité et de réplication du VIH-2, nous avons étudié les caractéristiques immuno-virologiques des réservoirs du VIH-2 chez des patients VIH-2+ (ANRS-CO-05). Quatorze sujets VIH-2+ ont été comparés à des donneurs sains. Les cellules sanguines ont été triées en sept sous-populations (monocytes, T CD4 activés et au repos (TN, TCM, TTM, TEM)). L’ADN du VIH-2 a été quantifié par PCR et détecté chez 4 sujets : dans les TTM et les TCM mais pas dans les monocytes. L’analyse phénotypique des cellules NK a été réalisée par cytométrie en flux (24 sujets, ANRS-CO-05). Les cellules NK des VIH-2+ avaient : une expression de NKp30 diminuée, une augmentation de la production d'IFN-g et une altération de la fonction cytolytique en présence de B7-H6, comparés aux VIH-1+. L’expression des corécepteurs et des facteurs de transcription a été analysée, montrant des niveaux plus élevés chez les VIH-1 LTNP comparée aux VIH-2+, et aux donneurs sains pour la plupart des paramètres. Nous avons observé que l’expression de PD1 était plus importante sur les T CD4 des VIH-1 LTNP comparés aux donneurs sains. Nous avons montré que les faibles réservoirs du VIH-2 se distinguent des réservoirs du VIH-1 par l'absence d'infection monocytaire et par une infection limitée des TCM.HIV-2 and HIV-1 differ in pathogenicity. To better understand the mechanisms of HIV- 2 low pathogenicity and replication potential, we studied immune and virologic characteristics of HIV-2 reservoirs of infected patients (cohort ANRS CO-05). Fourteen HIV-2+ subjects from the ANRS CO-05 cohort were compared to healthy donors. Seven peripheral blood sub-populations were sorted by flow cytometry : Monocytes, activated and resting CD4 T cells (TN, TCM, TTM, TEM). Viral DNA was quantified in sorted cells by PCR. HIV-2 DNA was detected in TTM and TCM sub-populations of 4 patients but not in monocytes. The phenotype of peripheral NK cells was analyzed by flow cytometry (24 HIV-2 ANRS CO-05 cohort). HIV-2+ patients had lower proportions of NKp30+ NK cells, increased production of IFN-g and altered cytolytic function in the presence B7-H6 surface ligand, when compared to HIV-1+. The analysis of HIV-1 and HIV-2 co-receptors and transcription factors expression showed higher values in HIV-1LTNP (ANRS CO15) cohort, compared to HIV-2+ and healthy donors. We also found that the PD1 expression by CD4 T cells was higher in HIV-1LTNP subjects compared to healthy donors, though not correlated to the HIV-1 plasma load. Her we show that HIV-2 reservoirs are distinct from the HIV-1 reservoirs by the absence of monocytic infection and a limited TCM infection

Initiating antiretroviral treatment early in infancy has long-term benefits on the HIV reservoir in late childhood and adolescence

International audienceBackgroundEarly combined antiretroviral therapy (cART) limits the total HIV-DNA load in children. However, data on its impact in older children and adolescents remain scarce. This study aims to compare HIV reservoirs in children (5-12 years) and adolescents (13-17 years) who started cART before 6 months (early (E-)group) or after 2 years old (late (L-)group). MethodsThe ANRS-EP59-CLEAC study prospectively enrolled 76 HIV-1 perinatally-infected patients who reached HIV-RNA<400 copies/mL less than 24 months after cART initiation, regardless of subsequent viral suppression (E-group: 27 children, 9 adolescents; L-group: 19 children, 21 adolescents). Total and integrated HIV-DNA were quantified in blood and in CD4+ T cell subsets. A substudy assessed HIV reservoir inducibility after ex vivo peripheral blood mononuclear cells (PBMCs) stimulation.ResultsTotal HIV-DNA levels were lower in early- than late-treated patients (Children: 2.14 vs 2.87 log cp/million PBMCs, p<0.0001; Adolescents: 2.25 vs 2.74log, p<0.0001). Low reservoir was independently associated with treatment precocity, protective HLA and low cumulative viremia since cART initiation. The 60 participants with undetectable integrated HIV-DNA started cART earlier than the other patients (4 vs 54 months, p=0.03). In those with sustained virological control, transitional memory and effector memory CD4+T cells were less infected in the E-group than in the L-group (p=0.03 and 0.02, respectively). Viral inducibility of reservoir cells after normalization to HIV-DNA levels was similar between the groups.ConclusionsEarly cART results in a smaller blood HIV reservoir until adolescence, but all tested participants had an inducible reservoir. This deserves cautious consideration for HIV remission strategies

Surveillance of transmitted HIV-1 antiretroviral drug resistance in the context of decentralized HIV care in Senegal and the Ebola outbreak in Guinea

Abstract Objectives Disruption in HIV care provision may enhance the development and spread of drug resistance due to inadequate antiretroviral therapy. This study thus determined the prevalence of HIV-1 transmitted drug resistance (TDR) in settings of decentralized therapy and care in Senegal and, the Ebola outbreak in Guinea. Antiretroviral-naïve patients were enrolled following a modified WHO TDR Threshold Survey method, implemented in Senegal (January–March 2015) and Guinea (August–September 2015). Plasma and dried blood spots specimens, respectively from Senegalese (n = 69) and Guinean (n = 50) patients, were collected for direct sequencing of HIV-1 pol genes. The Stanford Calibrated Population Resistance program v6.0 was used for Surveillance Drug Resistance Mutations (SDRMs). Results Genotyping was successful from 54/69 (78.2%) and 31/50 (62.0%) isolates. In Senegal, TDR prevalence was 0% (mean duration since HIV diagnosis 4.08 ± 3.53 years). In Guinea, two patients exhibited SDRMs M184V (NRTI), T215F (TAM) and, G190A (NNRTI), respectively. TDR prevalence at this second site, however, could not be ascertained because of low sample size. Phylogenetic inference confirmed CRF02_AG predominance in Senegal (62.96%) and Guinea (77.42%). TDR prevalence in Senegal remains extremely low suggesting improved control measures. Continuous surveillance in both settings is mandatory and, should be done closest to diagnosis/transmission time and with larger sample size

AIDS Res Hum Retroviruses

Human immunodeficiency viruses induce rare attenuated diseases due either to HIV-1 in the exceptional long-term nonprogressors (LTNPs) or to HIV-2 in West Africa. To better understand characteristics of these two disease types we performed a multiplex comparative analysis of cell activation, exhaustion, and expression of coreceptors and restriction factors in CD4 T cells susceptible to harbor those viruses. We analyzed by flow cytometry the expression of HLA-DR, PD1, CCR5, CXCR6, SAMHD1, Blimp-1, and TRIM5 alpha on CD4 T cell subsets from 10 HIV-1(+) LTNPs and 14 HIV-2(+) (12 nonprogressors and 2 progressors) of the ANRS CO-15 and CO-5 cohorts, respectively, and 12 HIV- healthy donors (HD). The V3 loop of the HIV-1 envelope from 6 HIV-1+ LTNPs was sequenced to determine the CXCR6-binding capacity. Proportions of HLA-DR+ and PD1+ cells were higher in memory CD4 T subsets from HIV-1 LTNPs compared with HIV-2 and HD. Similar findings were observed for CCR5+ cells although limited to central-memory CD4 T cell (TCM) and follicular helper T cell subsets, whereas all major subsets from HIV-1 LTNPs contained less CXCR6+ cells compared with HIV-2. All six V3 loop sequences from HIV-1 LTNPs contained a proline at position 326. Proportions of SAMHD1+ cells were higher in all resting CD4 T subsets from HIV-1 LTNPs compared with the other groups, whereas Blimp-1+ and Trim5 alpha+ cells did not differ. The CD4 T cell subsets from HIV-1 LTNPs differ from those of HIV-2-infected subjects by higher levels of activation, exhaustion, and SAMHD1 expression that can reflect the distinct patterns of host/virus relationships