Bénédicte Roquebert

Hôpital Bichat - Claude-Bernard (Hôpitaux Universitaires Paris Nord Val de Seine), Lutetia Parisorum, Île-de-France, France

Are you Bénédicte Roquebert?

Claim your profile

Publications (31)

  • [Show abstract] [Hide abstract] ABSTRACT: Among 141 HIV-HBV-coinfected patients treated with tenofovir in our centre, 87% were good-responders to therapy. Seven patients showed a delayed response to tenofovir. The present study was performed to evaluate the quasispecies variability and the in vitro drug susceptibility to approved antiviral drugs of HBV genomes directly isolated from patients' sera. After purification of DNA from serum samples, full-length HBV DNA was amplified by PCR, cloned and sequenced. Drug sensitivity of HBV strains isolated from four delayed responders and five good-responders was assessed and compared to a wild-type HBV strain after transfection of the full genome into HepG2 cells. Delayed responders, compared with good responders, showed a higher incidence of lamivudine-resistant mutations (71% and 35%, respectively; P=0.021) and a higher proportion of HBV genotype G (57% versus 16%, respectively; P=0.026). Clonal analysis demonstrated a higher variability of HBV quasispecies in delayed reponders than in good responders. In vitro analysis showed a lower efficacy of adefovir and tenofovir in delayed reponders. Furthermore, HBV genotype G strains showed a mild to weak susceptibility to tenofovir. The reason for the slow decline in HBV DNA level observed during therapy in delayed responders is not clear. Delayed responders showed higher quasispecies variability, a higher proportion of HBV genotype G and a mild in vitro decreased susceptibility to tenofovir and adefovir. A combination of these factors in heavily treatment-experienced HIV-infected patients could explain the lower tenofovir activity. These patients must be closely monitored to prevent prospective emergence of resistance to approved antiviral drugs.
    Article · Jan 2012 · Antiviral therapy
  • [Show abstract] [Hide abstract] ABSTRACT: To evaluate the early virological response (EVR) to combined tenofovir-lamivudine or emtricitabine regimen in HBV/HIV-co-infected patients and the long-term efficacy of tenofovir. In this retrospective monocentric study, among the 166 HIV/HBV-co-infected patients regularly followed from 2003 to 2008 at Bichat Claude Bernard Hospital, 61 patients had received, either de novo combination therapy with tenofovir and lamivudine or emtricitabine (group I, n = 15) or add-on tenofovir to lamivudine therapy (group II, n = 46). The HBV polymerase region was sequenced and analysed for all patients with available samples. All 15 group I patients achieved EVR vs 32 (82%) of group II patients (P = 0.15). Seven adherent group II patients met criteria for primary non-response, but achieved delayed response (DR) to therapy. In these seven patients, when compared with the 39 group II patients, there was a trend to longer duration of lamivudine pre-treatment and to higher rate of lamivudine-resistant mutants; and HBV genotype-G proportion was higher (P = 0.026). No virological breakthrough occurred after a median of 46 months follow up. In these HBV/HIV-co-infected patients, first-line HBV therapy with tenofovir and emtricitabine or lamivudine was associated with EVR. However, DR to tenofovir was observed in 15% of patients who added tenofovir to lamivudine therapy, of whom four of seven (57%) had genotype G-HBV infection. No resistance was evidenced after 46 months of therapy even in patients with DR to tenofovir. At last, a good renal safety profile of TDF was observed after a median follow-up of 4 years of therapy.
    Article · Aug 2011 · Liver international: official journal of the International Association for the Study of the Liver
  • Source
    Charlotte Charpentier · Bénédicte Roquebert · Olivier Delelis · [...] · Diane Descamps
    [Show abstract] [Hide abstract] ABSTRACT: We studied seven heavily pretreated HIV-2-infected patients exhibiting a virological failure while receiving a salvage raltegravir-containing regimen. At the time of virological failure, different resistance genetic pathways were observed: T97A-Y143C, Q148K, Q148R, G140S-Q148R, E92Q-Y143R-N155H, and T97A-N155H. Thus, despite a 40% difference in integrase genes between HIV-1 and HIV-2, the genetic pathways leading to raltegravir resistance are similar. Copyright © 2011, American Society for Microbiology. All Rights Reserved.
    Full-text Article · Dec 2010 · Antimicrobial Agents and Chemotherapy
  • Charlotte Charpentier · Bénédicte Roquebert · Céline Colin · [...] · Diane Descamps
    [Show abstract] [Hide abstract] ABSTRACT: ANRS 139 TRIO trial was a phase II noncomparative trial that evaluated in highly experienced patients, a combination of raltegravir, etravirine and darunavir boosted with ritonavir. We analyzed emergence of resistant viruses at the time of virological failure and investigated the impact of baseline integrase polymorphisms on virological failure occurrence. Bulk sequencing of protease, reverse transcriptase and integrase genes was performed for 103 patients at baseline and 14 patients at the time of virological failure. Additionally, integrase clonal analyses were performed at baseline and at virological failure in patients with successful integrase gene amplification. Impact of baseline integrase polymorphisms on virological failure occurrence was analyzed using Fisher exact and Wilcoxon tests. In the 14 failing patients median viral load at virological failure was 90 copies/ml (interquartile range = 60-783). Emergence of darunavir and etravirine resistance mutations was observed at virological failure in only one and three patients, respectively. Raltegravir resistance mutations were found neither at baseline nor at the time of virologic failure. Integrase clonal analyses showed neither the presence nor the selection of minority variants carrying raltegravir resistance mutations at baseline or at virological failure. No impact of baseline integrase polymorphisms was observed on virological failure either at week 24 or at week 48. Virological failure occurred in a small proportion of patients with low viral load. No raltegravir resistance mutations were observed using bulk sequencing or clonal analyses, and darunavir and etravirine resistance-associated mutations were detected in only one and three patients, respectively at virological failure. No impact of baseline integrase polymorphism was observed on virological failure occurrence.
    Article · Nov 2010 · AIDS (London, England)
  • [Show abstract] [Hide abstract] ABSTRACT: We evaluated the performance of the prototype Cobas AmpliPrep/Cobas TaqMan HIV-1 test, version 2.0, using prospective and archived clinical samples initially underquantitated by the Cobas AmpliPrep/Cobas TaqMan HIV-1 test. The performance of the new test was significantly improved, and the majority of the underquantitation observed with the first-version test was eliminated.
    Article · Apr 2010 · Journal of clinical microbiology
  • B. Roquebert · J. Leleu
    [Show abstract] [Hide abstract] ABSTRACT: Twenty-five years after the beginning of HIV antiretroviral therapy, reverse transcriptase inhibitors (RTIs) remain the backbone of HAART. Unfortunately, the emergence of drug resistance can undermine the benefits of this potent treatment. As regards RTIs resistance, the vast majority of well-described resistance-associated mutations are clustered in the N-terminal polymerase domain of HIV-1 reverse transcriptase. However, this enzyme is not limited to a single domain and mutations observed at considerable distance from this location can likewise affect drug susceptibility by themselves or in association with "classical" mutations. Some authors suggest that mutations in the C-terminal, connection and RNase H, domains are associated with antiretroviral therapy. The potential clinical significance of these mutations is presented in this review. In the second part, we report the plausible biochemical mechanisms explaining how these mutations can contribute to affect drug susceptibility and viral replication capacity. Overall, sequencing the entire reverse transcriptase in order to understand resistance should be useful due to the presence of these mutations outside of the polymerase domain, but further investigations are still warranted to determine their impact in clinical practice.
    Article · Mar 2010 · Virologie
  • Benedicte Roquebert · Juliette Leleu
    [Show abstract] [Hide abstract] ABSTRACT: Twenty-five years after the beginning of HIV antiretroviral therapy, reverse transcriptase inhibitors (RTIs) remain the backbone of HAART. Unfortunately, the emergence of drug resistance can undermine the benefits of this potent treatment. As regards RTIs resistance, the vast majority of well-described resistance-associated mutations are clustered in the N-terminal polymerase domain of HIV-1 reverse transcriptase. However, this enzyme is not limited to a single domain and mutations observed at considerable distance from this location can likewise affect drug susceptibility by themselves or in association with "classical" mutations. Some authors suggest that mutations in the C-terminal, connection and RNase H, domains are associated with antiretroviral therapy. The potential clinical significance of these mutations is presented in this review. In the second part, we report the plausible biochemical mechanisms explaining how these mutations can contribute to affect drug susceptibility and viral replication capacity. Overall, sequencing the entire reverse transcriptase in order to understand resistance should be useful due to the presence of these mutations outside of the polymerase domain, but further investigations are still warranted to determine their impact in clinical practice.
    Article · Jan 2010 · Virologie
  • Source
    [Show abstract] [Hide abstract] ABSTRACT: The introduction of 2 or 3 fully active drugs in human immunodeficiency virus (HIV)-infected patients receiving failing antiretroviral therapy is a key determinant of subsequent treatment efficacy. The aim of this study was to assess the safety and efficacy of a regimen containing raltegravir, etravirine, and darunavir/ritonavir for treatment-experienced patients infected with multidrug-resistant HIV. Patients enrolled in this phase II, noncomparative, multicenter trial were naive to the investigational drugs and had plasma HIV RNA levels >1000 copies/mL, a history of virologic failure while receiving nonnucleoside reverse-transcriptase inhibitors (NNRTI), > or =3 primary protease inhibitor and nucleoside reverse transcriptase inhibitor (NRTI) mutations, and < or =3 darunavir and NNRTI mutations. The primary end point was the proportion of patients with plasma HIV RNA levels <50 copies/mL at 24 weeks. A total of 103 patients enrolled in the study. At baseline, genotypic resistance profiles showed a median of 4 primary protease inhibitor mutations, 1 NNRTI mutation, and 6 NRTI mutations. In addition to the investigational drugs, 90 patients (87%) received optimized background therapy that included NRTIs (86 patients) or enfuvirtide (12 patients). At week 24, 90% of patients (95% confidence interval, 85%-96%) had an HIV RNA level <50 copies/mL. At week 48, 86% (95% confidence interval, 80%-93%) had an HIV RNA level <50 copies/mL. The median CD4 cell count increase was 108 cells/mm(3). Grade 3 or 4 clinical adverse events were reported in 15 patients (14.6%). Only 1 patient discontinued the investigational antiretroviral regimen, because of an adverse event. In patients infected with multidrug-resistant virus who have few remaining treatment options, the combination of raltegravir, etravirine, and darunavir/ritonavir is well tolerated and is associated with a rate of virologic suppression similar to that expected in treatment-naive patients.
    Full-text Article · Nov 2009 · Clinical Infectious Diseases
  • [Show abstract] [Hide abstract] ABSTRACT: The aim of the study was to identify a pattern of protease gene mutations associated with the virological response to darunavir/ritonavir-based regimens. We analysed 153 treatment-experienced patients receiving a darunavir/ritonavir salvage regimen as a sole protease inhibitor (PI). Virological response was defined as an HIV-1 RNA load of <200 copies/mL at month 3. The impact of individual protease gene mutations on the virological response to darunavir/ritonavir was examined, and the combination of mutations most strongly associated with the virological response was identified. The baseline median HIV RNA level was 4.7 log(10) copies/mL and the median CD4 cell count was 142 cells/mm(3). At month 3, 55% of patients had a virological response and the median fall in viral load from baseline was 1.7 log(10) copies/mL. All the patients had detectable darunavir concentrations at month 3. Cochran-Armitage procedure identified eight mutations with a negative impact on the virological response, namely K14R, K20I, E34Q, I47V, I54M, K55R, T74P and I84V; and two mutations (E35D and V82A) with a positive impact. In multivariate analyses, our genotypic scores were highly predictive of the virological response at month 3, along with the baseline plasma viral load and enfuvirtide co-prescription to enfuvirtide-naive patients. Among the eight mutations with a negative impact on the virological response, I47V, I54M, T74P and I84V were previously described as darunavir resistance-associated mutations. Some PI resistance mutations had a positive impact on the virological response. These findings might help to explain the potency of darunavir/ritonavir on PI-resistant HIV.
    Article · Jan 2009 · Journal of Antimicrobial Chemotherapy
  • Bénédicte Roquebert · Laurent Blum · Gilles Collin · [...] · Diane Descamps
    Article · Nov 2008 · AIDS (London, England)
  • Source
    B Roquebert · F Damond · G Collin · [...] · D Descamps
    [Show abstract] [Hide abstract] ABSTRACT: We investigated the in vitro phenotypic susceptibility of HIV-2 isolates from integrase inhibitor (INI)-naive patients to INIs and its relation to HIV-2 integrase gene polymorphism. We determined the phenotypic susceptibility to raltegravir and elvitegravir of co-cultured isolates obtained from the HIV-2 ROD reference strain and from 14 clinical isolates. IC(50) values were compared with those for HIV-1 reference strains. HIV-2 integrase gene polymorphism was assessed in isolates from 52 INI-naive patients enrolled in the French HIV-2 cohort. Median raltegravir and elvitegravir IC(50) values for the 14 clinical HIV-2 isolates were 2.4 and 0.7 nM, respectively, and were similar to those observed for HIV-2 ROD and HIV-1 reference strains. Overall, 38% of HIV-2 integrase amino acids were polymorphic. The catalytic triad DDE and the HHCC and RKK motifs were fully conserved, at the same genomic positions as described in HIV-1. In subtype B isolates, the total length of the integrase gene varied, owing to the presence of stop codons at positions 288, 294, 297 and 302. Fourteen of the positions associated with substitutions conferring INI resistance in HIV-1 were polymorphic in HIV-2. Despite 40% heterogeneity between the HIV-1 and HIV-2 integrase genes, the phenotypic susceptibility of clinical HIV-2 isolates to INIs was similar to that of HIV-1. This new class of antiretroviral drugs thus represents a novel therapeutic possibility for HIV-2-infected patients who otherwise have few treatment options.
    Full-text Article · Sep 2008 · Journal of Antimicrobial Chemotherapy
  • [Show abstract] [Hide abstract] ABSTRACT: An abstract is unavailable. This article is available as HTML full text and PDF.
    Article · Jun 2008 · AIDS (London, England)
  • B Roquebert · F Damond · F Brun-Vézinet · D Descamps
    [Show abstract] [Hide abstract] ABSTRACT: Human immunodeficiency viruses HIV-1 and HIV-2 are the results of multi-interspecies transmissions from simian virus to humans. HIV-1 viruses are very divergent and are classified in three groups: M, N and O. The group M is subdivided in nine subtypes and numerous Circulating Recombinant Forms. In 1996, protease inhibitors and HAART disposal have modified the prognostic of the HIV infection. However, one of the major problems is the emergence of antiretroviral resistance. A major advance from the last year is the access to antiretroviral in resources limited countries. On the other hand, the development of a vaccine is today hypothetic.
    Article · Jun 2008 · Pathologie Biologie
  • Bénédicte Roquebert · Anne-Geneviève Marcelin
    [Show abstract] [Hide abstract] ABSTRACT: Some recent studies have underlined the role of the entire reverse transcriptase (RT) and in particular its carboxy-terminal domain from amino acid 427 to amino acid 560 [the ribonuclease H (RNAse H) domain] in resistance to nucleoside RT inhibitors (NRTIs). RNAse H is implicated in catalysing the degradation of the RNA strand during conversion of the viral genome into double-stranded DNA. It has been shown, by site-directed mutagenesis, that amino acid substitutions in the RNAse H domain could affect the binding enzyme/substrate, resulting in a decrease in the RNAse H activity. For example, mutations at positions 478, 539 and 549 led to a slowing down in the degradation of the RNA strand. In vitro, the mutations H539N and D549N decreased the frequency of RT template-switching and, thereby, increased the time for excision of incorporated NRTIs, and thus enhanced NRTI resistance. It has been confirmed in vivo that mutations at position 558 were statistically associated with a number of thymidine analogue mutations in a study including 144 HIV-1 patients, suggesting that it could be an accessory mutation that could reinforce NRTI resistance. This article highlights evidence that mutations in RNAse H can enhance NRTI resistance, suggesting that phenotypic and genotypic analyses of clinical samples including the entire HIV-1 RT and in particular the RNAse H domain are now required to better characterize the in vivo role of the RNAse H mutations on susceptibility and response to NRTIs in HIV-1-infected patients.
    Article · Jun 2008 · Journal of Antimicrobial Chemotherapy
  • Source
    Delphine Desbois · Bénédicte Roquebert · Gilles Peytavin · [...] · Diane Descamps
    [Show abstract] [Hide abstract] ABSTRACT: We determine phenotypic susceptibility of human immunodeficiency virus type 2 (HIV-2) isolates to amprenavir, atazanavir, darunavir, indinavir, lopinavir, nelfinavir, saquinavir, and tipranavir. Saquinavir, lopinavir, and darunavir are potent against wild-type HIV-2 isolates and should be preferred as first-line options for HIV-2-infected patients. Other protease inhibitors are less active against HIV-2 than against HIV-1.
    Full-text Article · May 2008 · Antimicrobial Agents and Chemotherapy
  • Florence Damond · Sylvie Lariven · Benedicte Roquebert · [...] · Sophie Matheron
    [Show abstract] [Hide abstract] ABSTRACT: An abstract is unavailable. This article is available as HTML full text and PDF.
    Article · Apr 2008 · AIDS (London, England)
  • Source
    Full-text Article · Mar 2008 · Journal of clinical microbiology
  • B. Roquebert · D. Descamps
    [Show abstract] [Hide abstract] ABSTRACT: Two inhibitors of the HIV-1 integrase are in late stage clinical development in studies including HIV-1 treatment-experienced patients. Efficacy results of phase III study for raltegravir and phase II study for elvitegravir are excellent. But, as with other antiretroviral classes, drug resistance has been shown to occur with the integrase inhibitors. To date, 42 mutations within the integrase gene have been associated in vitro with resistance. Each class of integrase inhibitors has a distinct mechanism of action and carries their respective resistance profile. Polymorphism and clinical studies reinforced the importance of some relevant mutations significantly associated with virologic failure in patients under drug regimen including integrase inhibitors. All of these results led to include a resistance profile about raltegravir compound in the ANRS resistance genotypic algorithm in July 2007. Nevertheless, other larger studies are needed to answer questions about cross resistance patterns and time to developing resistance in HIV-1 infected patients and treated with these new drugs.
    Article · Dec 2007 · Virologie
  • Source
    F Damond · B Roquebert · A Bénard · [...] · D Descamps
    [Show abstract] [Hide abstract] ABSTRACT: We compared plasma viral load values obtained with COBAS AMPLICOR human immunodeficiency virus type 1 (HIV-1) MONITOR version 1.5 and with COBAS TaqMan HIV-1 assays. Mean values were 4.2 and 2.9 log10 copies/ml, respectively, showing the lack of agreement between the two assays.
    Full-text Article · Nov 2007 · Journal of Clinical Microbiology
  • Isabelle Malet · Bénédicte Roquebert · Cécile Dalban · [...] · Anne-Geneviève Marcelin
    [Show abstract] [Hide abstract] ABSTRACT: The sequence variability in the protease and in the 5 Gag cleavage sites (CS) were explored to look for eventual associations between the mutations. Moreover, we have evaluated associations between the Gag region sequence and the virological response to Protease Inhibitors (PI). The protease and the 5 Gag CS sequences from 98 PI-experienced patients were sequenced and compared to the HXB2 reference sequence. Sixty patients, treated by Saquinavir plus Ritonavir, were studied to evaluate the clinical impact of the Gag region variability. The relationship between 63 protease mutations and 21 Gag CS mutations were explored. Two patterns of mutations in the protease were identified: (M46I/L, I54V, V82A/T/F) was associated to the A431V and (K20I/R/M, L89M/I) to the S373Q and L449P. None of the Gag CS mutations resulting from PI treatment was associated to the virological response to SQV/r. On the other hand, the S373P mutation had a negative impact on the virological response that remained statistically significant in a multivariate analysis after adjustment on the number of PI resistance mutations. These results evoke the pertinence to introduce some mutations found in the Gag CS in the algorithms used for the interpretation of resistance testing.
    Article · May 2007 · The Journal of infection