Bonaventura Clotet

Publications of Bonaventura Clotet

• A surprising evolution from oral human papillomavirus 16 infection to lymph node metastasis of tonsilar squamous cell carcinoma in an HIV-infected patient.

  Authors: Guillem Sirera, Sebastián Videla, Maripaz Cañadas, Laila Darwich, Emanuela d'Angelo, Bonaventura Clotet

  AIDS (London, England). 05/2012; 26(8):1044-5.

• HIV-1 Capture and Antigen Presentation by Dendritic Cells: Enhanced Viral Capture Does Not Correlate with Better T Cell Activation.

  Authors: Maria T Rodriguez-Plata, Alejandra Urrutia, Sylvain Cardinaud, Maria J Buzón, Nuria Izquierdo-Useros, Julia G Prado, Maria C Puertas, Itziar Erkizia, Pierre-Grégoire Coulon, Samandhy Cedeño, Bonaventura Clotet, Arnaud Moris, Javier Martinez-Picado

  Journal of immunology (Baltimore, Md. : 1950). 05/2012;

  During HIV-1 infection, dendritic cells (DC) facilitate dissemination of HIV-1 while trying to trigger adaptive antiviral immune responses. We examined whether increased HIV-1 capture in DC maturedDuring HIV-1 infection, dendritic cells (DC) facilitate dissemination of HIV-1 while trying to trigger adaptive antiviral immune responses. We examined whether increased HIV-1 capture in DC matured with LPS results in more efficient Ag presentation to HIV-1-specific CD4(+) and CD8(+) T cells. To block the DC-mediated trans-infection of HIV-1 and maximize Ag loading, we also evaluated a noninfectious integrase-deficient HIV-1 isolate, HIV(NL4-3ΔIN). We showed that higher viral capture of DC did not guarantee better Ag presentation or T cell activation. Greater HIV(NL4-3) uptake by fully LPS-matured DC resulted in higher viral transmission to target cells but poorer stimulation of HIV-1-specific CD4(+) and CD8(+) T cells. Conversely, maturation of DC with LPS during, but not before, viral loading enhanced both HLA-I and HLA-II HIV-1-derived Ag presentation. In contrast, DC maturation with the clinical-grade mixture consisting of IL-1β, TNF-α, IL-6, and PGE(2) during viral uptake only stimulated HIV-1-specific CD8(+) T cells. Hence, DC maturation state, activation stimulus, and time lag between DC maturation and Ag loading impact HIV-1 capture and virus Ag presentation. Our results demonstrate a dissociation between the capacity to capture HIV-1 and to present viral Ags. Integrase-deficient HIV(NL4-3ΔIN) was also efficiently captured and presented by DC through the HLA-I and HLA-II pathways but in the absence of viral dissemination. HIV(NL4-3ΔIN) seems to be an attractive candidate to be explored. These results provide new insights into DC biology and have implications in the optimization of DC-based immunotherapy against HIV-1 infection.

• Ritonavir boosting dose reduction from 100 to 50 mg does not change the atazanavir steady-state exposure in healthy volunteers.

  Authors: Javier A Estévez, José Moltó, Laura Tuneu, Samandhy Cedeño, Rosa M Antonijoan, Maria A Mangues, Bonaventura Clotet, Pere Domingo, Montse Puntes, Manuel J Barbanoj, Marta Valle

  The Journal of antimicrobial chemotherapy. 05/2012;

  OBJECTIVES: To evaluate the pharmacokinetics, tolerability and safety of 300 mg of atazanavir boosted with 100 or 50 mg of ritonavir, both once daily, at steady state. METHODS: This was aOBJECTIVES: To evaluate the pharmacokinetics, tolerability and safety of 300 mg of atazanavir boosted with 100 or 50 mg of ritonavir, both once daily, at steady state. METHODS: This was a single-blind, multiple-dose, crossover, sequence-randomized trial. Thirteen healthy HIV-1-negative men received witnessed once-daily doses of atazanavir (300 mg) and 100 or 50 mg of ritonavir for 10 days (15 day washout). Atazanavir and ritonavir plasma concentrations were determined for 24 h on day 10. Log-transformed individual pharmacokinetic parameters were compared between treatments (analysis of variance); the difference between treatments on the log scale and 95% CIs were calculated. Fasting cholesterol, triglycerides, glucose and bilirubin plasma levels were measured at the beginning and end of each period and compared (Wilcoxon signed rank test). Gastrointestinal symptoms and other events were recorded. RESULTS: Ritonavir C(max) and the AUC(0-24) were lower after the 50 mg booster dose than after 100 mg [geometric mean ratio (GMR) (95% CI), 0.40 (0.31-0.51) and 0.35 (0.29-0.42), respectively]. No differences were observed in atazanavir exposure with 50 or 100 mg of ritonavir [GMR C(max) (95% CI), 1.00 (0.79-1.28); GMR AUC(0-24) (95% CI), 0.98 (0.79-1.21)]. Atazanavir trough concentration was >0.15 mg/L in all volunteers. Total and low-density lipoprotein cholesterol increased 0.40 mM (P = 0.01) and 0.37 mM (P = 0.003) from their corresponding baseline value during the 100 mg dosing period; there were no significant changes on 50 mg. Mild increases in bilirubin were detected on day 10 after both treatments without differences between treatments. CONCLUSIONS: In spite of higher exposure to ritonavir with 100 mg, atazanavir exposure was equivalent; the lipid profile was better under the lower booster dose (50 mg).

• [Costs and cost-efficacy analysis of the preferred treatments by GESIDA/National plan for AIDS for the initial antiretroviral therapy in adult human Immunodeficiency virus (HIV) infected patients in 2012.]

  Authors: Antonio Javier Blasco, José Ramón Arribas, Vicente Boix, Bonaventura Clotet, Pere Domingo, Juan González-García, Hernando Knobel, Juan Carlos López, Josep M Llibre, Fernando Lozano, José M Miró, Daniel Podzamczer, Juan Miguel Santamaría, Montserrat Tuset, Laura Zamora, Pablo Lázaro, Josep M Gatell

  Enfermedades infecciosas y microbiologia clinica. 04/2012;

  INTRODUCTION: The GESIDA and National AIDS Plan panel of experts propose «preferred regimens» of antiretroviral treatment (ART) as initial therapy in HIV infected patients for 2012. The objective ofINTRODUCTION: The GESIDA and National AIDS Plan panel of experts propose «preferred regimens» of antiretroviral treatment (ART) as initial therapy in HIV infected patients for 2012. The objective of this study is to evaluate the costs and the efficiency of initiating treatment with these «preferred regimens». METHODS: Economic assessment of costs and efficiency (cost/efficacy) using decision tree analysis model. Efficacy was defined as the probability of having a viral load <50 copies/ml at week 48, in an intention-to-treat analysis. Cost of initiating treatment with an ART regime was defined as the costs of ART and all its consequences (adverse effects, changes of ART regime, and drug resistance analyses) during the first 48 weeks. The perspective of the analysis is that of the National Health System, considering only differential direct costs: ART (official prices), management of adverse effects, studies of resistance and determination of HLA B*5701. The setting is Spain and the costs are those of 2012. A sensitivity deterministic analysis was conducted, building three scenarios for each regime: baseline, most favourable, and most unfavourable cases. RESULTS: In the baseline case scenario, the cost of initiating treatment ranges from 6,895 euros for TDF/FTC+NVP to 12,067 euros for TDF/FTC+RAL. The efficacy ranges between 0.66 for ABC/3TC+LPV/r and 0.87 for TDF/FTC+RAL. Efficiency, in terms of cost/efficacy, varies between 9,387 and 13,823 euros per responder at 48 weeks, for TDF/FTC/EFV and TDF/FTC+RAL, respectively. In the most unfavourable scenario, the most efficient regime is TDF/FTC+NVP (9,742 per responder). CONCLUSION: Considering the official prices of ART, the most efficient regimens are TDF/FTC/EFV (baseline case and most favourable scenarios), and TDF/FTC+NVP (most unfavourable scenario).

• Effect of Liver Fibrosis on Long-Term Mortality in HIV/HCV Coinfected Individuals Who Are Evaluated to Receive Interferon Therapies in the HAART Era.

  Authors: Ramon Sanmartín Sentañes, Elena De Felipe, Jordi Tor, Arantza Sanvisens, Eva Barluenga, Elisenda Martínez, Robert Muga, Antoni Jou, Isabel Ojanguren, Juan Jose Lopez, Bonaventura Clotet, Cristina Tural

  AIDS research and human retroviruses. 03/2012;

  Objective: to evaluate the factors associated with overall mortality and liver decompensation in HIV and HCV coinfected patients who are evaluated to receive HCV antiviral therapy with a known liverObjective: to evaluate the factors associated with overall mortality and liver decompensation in HIV and HCV coinfected patients who are evaluated to receive HCV antiviral therapy with a known liver histological fibrosis stage. Design: Prospective cohort study Methods: 387 Consecutive HIV/HCV coinfected patients attending an outpatient clinical unit between January 1997 and December 2007 who fulfilled criteria to be treated with interferon and to whom liver biopsy was performed, were included and followed every 6 months from time of liver biopsy to death or December 2008. Results: follow-up period was 6.2 years (IQR:3.5-9.2).Median age at time of liver biopsy: 38 years, 73% men, 29% with advanced liver fibrosis (F3-F4), CD4 cell count: 556 cells/mm3, 72% with HIV RNA <400 copies/mL, mean CD4 Nadir:207 cell/mm3, 21% had an AIDS previous diagnosis, 92% were on antiretroviral therapy. During follow-up 48% underwent HCV antiviral therapy, with a sustained virological response in 33%. Overall mortality rate and the incidence of liver decompensation or liver-related death was 1.17 and 0.72 per 100 patients-year respectively. End stage liver disease (9/28 patients) and non-AIDS related cancer (6/28) were the main causes of death. F3-F4 (HR:3.74, 95% CI: 1.69-8.26, p=0.001) and previous AIDS diagnosis (HR:3.04, 95% CI:1.36-6.81) were the factors independently associated with death. Mortality rate in patients who received and who did not receive HCV antiviral therapy were 0.44 and 2.04 per 100 patients-year respectively (p=0.003). Conclusions: Besides the low mortality rate observed, HIV/HCV coinfected patients with poor predictors of survival are candidates for intensive clinical management.

• Effect of Milk Thistle on the Pharmacokinetics of Darunavir/Ritonavir in HIV-Infected Patients.

  Authors: José Moltó, Marta Valle, Cristina Miranda, Samandhy Cedeño, Eugenia Negredo, Bonaventura Clotet

  Antimicrobial agents and chemotherapy. 03/2012;

  The aim of this open-label, fixed-sequence study was to investigate the potential of the botanical supplement milk thistle (silymarin) to interact with the boosted protease inhibitor combinationThe aim of this open-label, fixed-sequence study was to investigate the potential of the botanical supplement milk thistle (silymarin) to interact with the boosted protease inhibitor combination darunavir/ritonavir. Fifteen HIV-infected patients receiving antiretroviral therapy with darunavir/ritonavir (600/100 mg twice daily) for at least 4 weeks were included. Silymarin (150 mg every 8 hours) was added to the antiretroviral treatment from days 1 to 14. Darunavir concentrations in plasma were determined by high-performance liquid chromatography immediately before and 1, 2, 4, 6, 8, 10 and 12 hours after a morning dose of darunavir/ritonavir on day 0 and darunavir/ritonavir plus silymarin on day 14. Individual darunavir pharmacokinetic parameters were calculated by noncompartmental analysis and compared between days 0 and 14 by means of the geometric mean ratio (GMR) and its 90% confidence interval (CI). Median (interquartile range) age was 48 (44-50) years and body weight was 70 (65-84) kg. Silymarin was well tolerated and all participants completed the study. The GMR for darunavir coadministered with silymarin relative to darunavir alone was 0.86 (90% CI, 0.70-1.05) for the area under the concentration-time curve from 0 to 12 hours; 0.83 (90% CI, 0.80-0.98) for the maximum concentration, and 0.94 (90% CI, 0.73-1.19) for the concentration at the end of the dosing interval. In summary, coadministration of silymarin with darunavir/ritonavir seems to be safe in HIV-infected patients; no dose adjustment for darunavir/ritonavir seems necessary.

• 2-(Alkyl/Aryl)Amino-6-Benzylpyrimidin-4(3H)-ones as Inhibitors of Wild-Type and Mutant HIV-1: Enantioselectivity Studies.

  Authors: Dante Rotili, Alberta Samuele, Domenico Tarantino, Rino Ragno, Ira Musmuca, Flavio Ballante, Giorgia Botta, Ludovica Morera, Marco Pierini, Roberto Cirilli, Maxim B Nawrozkij, Emmanuel Gonzalez, Bonaventura Clotet, Marino Artico, José A Esté, Giovanni Maga, Antonello Mai

  Journal of medicinal chemistry. 03/2012;

  The single enantiomers of two pyrimidine-based HIV-1 non-nucleoside reverse transcriptase inhibitors, 1 (MC1501) and 2 (MC2082), were tested in both cellular and enzyme assays. In general, the RThe single enantiomers of two pyrimidine-based HIV-1 non-nucleoside reverse transcriptase inhibitors, 1 (MC1501) and 2 (MC2082), were tested in both cellular and enzyme assays. In general, the R forms were more potent than their S counterparts and racemates and (R)-2 was more efficient than (R)-1 and the reference compounds, with some exceptions. Interestingly, (R)-2 displayed a faster binding to K103N RT with respect to WT RT, while (R)-1 showed the opposite behavior.

• Mutations in the protease gene associated with virological failure to lopinavir/ritonavir-containing regimens.

  Authors: José R Santos, Josep M Llibre, Arkaitz Imaz, Pere Domingo, José A Iribarren, Ana Mariño, Celia Miralles, María J Galindo, Arelly Ornelas, Santiago Moreno, Jonathan M Schapiro, Bonaventura Clotet

  The Journal of antimicrobial chemotherapy. 03/2012;

  OBJECTIVES: To assess the most frequent resistance-associated mutations (RAMs) to lopinavir/ritonavir in a cohort of patients attended in daily practice. METHODS: We retrospectively identified 195OBJECTIVES: To assess the most frequent resistance-associated mutations (RAMs) to lopinavir/ritonavir in a cohort of patients attended in daily practice. METHODS: We retrospectively identified 195 multitreated subjects with virological failure. Patients were classified as follows: (i) 71 (36.4%) never received lopinavir/ritonavir (lopinavir/ritonavir naive); (ii) 75 (38.5%) had previously failed on lopinavir/ritonavir; and (iii) 49 (25.1%) were on lopinavir/ritonavir at failure. RAM patterns were assessed. Medians, IQRs, percentages, Kruskal-Wallis, χ(2) or Fisher's exact test, and multinomial logistic regression were used whenever appropriate. RESULTS: L10I/F, K20R, L24I, L33F, M36I, M46I/L, I47V, G48V, F53L, I54V, A71V, G73S, V82A, I84V and L90M (all with P ≤ 0.037) were protease RAMs overexpressed in patients with lopinavir/ritonavir failure. L10I, M36I, M46I, I54V, L63P, A71V, V82A, I84V and L90M were the most common in lopinavir/ritonavir-naive patients. Other IAS-USA RAMs for lopinavir/ritonavir (L10R/V, K20M, V32I, I47A, I50V, I54L/A/M/T/S, A71T, L76V and V82F/T/S) were not associated with previous or current failure to lopinavir/ritonavir. Lopinavir/ritonavir failure was associated with the number of protease RAMs (OR = 1.146, 95% CI = 1.287, 1.626), higher exposure to protease inhibitors, and the presence of E44D, L33F, I54V and I84V. CONCLUSIONS: In multitreated patients with previous or current lopinavir/ritonavir failure, some protease mutations are selected at significantly greater rates. L10I, M36I, I54V, L63P, A71V, V82A and L90M were found in >50% of cases. Thus, their presence should be expected when genotypic testing results are not available. The number of protease RAMs and higher prior exposures to protease inhibitors were significantly associated with lopinavir/ritonavir failure.

• Comparison of the f-HPV typing™ and Hybrid Capture II® assays for detection of high-risk HPV genotypes in cervical samples.

  Authors: María-Paz Cañadas, Vincenzo Cirigliano, Laila Darwich, Guillermo Sirera, Josep Coll, Bonaventura Clotet, Sebastian Videla

  Journal of virological methods. 03/2012;

  Human papillomavirus genotyping is being considered in cervical screening programs and for monitoring the effectiveness of HPV vaccination. Both approaches require access to fast, easy andHuman papillomavirus genotyping is being considered in cervical screening programs and for monitoring the effectiveness of HPV vaccination. Both approaches require access to fast, easy and high-throughput technology. The aim of this study was to compare a new commercial assay (f-HPV typing™) with the Hybrid Capture II® (HC2) to detect HPV infection. The F-HPV typing is a multiplex fluorescent PCR method recognizing E6 and E7 regions of 13 high-risk (HR) HPV types, the same set of HR-types targeted HC2 test. A subset of 157 cervical samples was tested with both assays. The percentage of positive HR-HPV DNA samples was 24% (37/155) by HC2 and 33% (49/155) by f-HPV typing. Concordant results were found in 133/155 (overall agreement, 85.8%; Cohen's kappa=0.65). The analytical sensitivity and specificity of f-HPV were 97.6 and 93, respectively. In conclusion, this study shows that the f-HPV assay provides a good alternative to HC2 to detect HPV infection, allowing simple and rapid HPV genotyping and detecting multiple infections.

• Evaluation of the Cytopathicity (Fusion/Hemifusion) of Patient-Derived HIV-1 Envelope Glycoproteins Comparing Two Effector Cell Lines.

  Authors: Francesc Cunyat, Marta Curriu, Silvia Marfil, Elisabet García, Bonaventura Clotet, Julià Blanco, Cecilia Cabrera

  Journal of biomolecular screening. 03/2012;

  HIV-1 envelope glycoprotein (Env) is a major determinant of viral pathogenicity. The evaluation of the biological properties of patient-derived envelopes by comparing two effector cell lines (293THIV-1 envelope glycoprotein (Env) is a major determinant of viral pathogenicity. The evaluation of the biological properties of patient-derived envelopes by comparing two effector cell lines (293T and HeLa) is reported. A standard cell-to-cell fusion assay was used to evaluate fusogenicity, whereas a coculture with CD4(+) cells was used to evaluate absolute cell loss, single cell death, and hemifusion events. Fusion and absolute cell loss assays showed that Env-expressing 293T and HeLa cells had different fusion efficiencies; fusion was magnified in 293T cells despite a significantly lower cell-surface Env expression. Conversely, gp41-mediated single cell death and hemifusion induced in CD4(+) cells by 293T-Env-positive cells were significantly lower than that induced by HeLa-Env-positive cells. These data showed that the effector cell line used in the in vitro assays is crucial, and a combination of assays is recommended to evaluate the biological properties of patient-derived envelope glycoproteins: preferentially, 293T-Env-positive cells for the evaluation of fusogenicity and HeLa-Env-positive cells for the evaluation of cell death parameters. The combination of assays described in our work could be a valuable tool for dual screenings of large collections of primary Envs or Env mutants and drugs acting on these Envs.

• Synthesis, Biological Activity, and ADME Properties of Novel S-DABOs/N-DABOs as HIV Reverse Transcriptase Inhibitors.

  Authors: Marco Radi, Mafalda Pagano, Luigi Franchi, Daniele Castagnolo, Silvia Schenone, Gianni Casaluce, Claudio Zamperini, Elena Dreassi, Giovanni Maga, Alberta Samuele, Encarna Gonzalo, Bonaventura Clotet, José A Esté, Maurizio Botta

  ChemMedChem. 03/2012;

  Previous studies aimed at exploring the SAR of C2-functionalized S-DABOs demonstrated that the substituent at this position plays a key role in the inhibition of both wild-type RT and drug-resistantPrevious studies aimed at exploring the SAR of C2-functionalized S-DABOs demonstrated that the substituent at this position plays a key role in the inhibition of both wild-type RT and drug-resistant enzymes, particularly the K103N mutant form. The introduction of a cyclopropyl group led us to the discovery of a potent inhibitor with picomolar activity against wild-type RT and nanomolar activity against many key mutant forms such as K103N. Despite its excellent antiviral profile, this compound suffers from a suboptimal ADME profile typical of many S-DABO analogues, but it could, however, represent a promising candidate as an anti-HIV microbicide. In the present work, a new series of S-DABO/N-DABO derivatives were synthesized to obtain additional SAR information on the C2-position and in particular to improve ADME properties while maintaining a good activity profile against HIV-1 RT. In vitro ADME properties (PAMPA permeation, water solubility, and metabolic stability) were also experimentally evaluated for the most interesting compounds to obtain a reliable indication of their plasma levels after oral administration.

• Rare LEDGF/p75 genetic variants in white long-term nonprogressor HIV+ individuals.

  Authors: Ester Ballana, Encarna Gonzalo, Eulàlia Grau, José A Iribarren, Bonaventura Clotet, José A Este

  AIDS (London, England). 02/2012; 26(4):527-8.

• The HR2 polymorphism N140I in the HIV-1 gp41 combined with the HR1 V38A mutation is associated with a less cytopathic phenotype.

  Authors: Francesc Cunyat, Silvia Marfil, Elisabet García, Valentina Svicher, Nuria Pérez-Alvárez, Marta Curriu, Carlo Federico Perno, Bonaventura Clotet, Julià Blanco, Cecilia Cabrera

  Retrovirology. 02/2012; 9:15.

  ABSTRACT: Resistance to the fusion inhibitor enfuvirtide (ENF) is achieved by changes in the gp41 subunit of the HIV envelope glycoprotein (Env). Specific ENF-associated mutational pathways correlateABSTRACT: Resistance to the fusion inhibitor enfuvirtide (ENF) is achieved by changes in the gp41 subunit of the HIV envelope glycoprotein (Env). Specific ENF-associated mutational pathways correlate with immunological recovery, even after virological failure, suggesting that the acquisition of ENF resistance alters gp41 pathogenicity. To test this hypothesis, we have characterized the expression, fusion capability, induction of CD4+ T cell loss and single CD4+ T cell death of 48 gp41 proteins derived from three patients displaying different amino acids (N, T or I) at position 140 that developed a V38A mutation after ENF-based treatment. In all cases, intra-patient comparison of Env isolated pre- or post-treatment showed comparable values of expression and fusogenic capacity. Furthermore, Env with either N or T at position 140 induced comparable losses of CD4+ T-cells, irrespective of the residue present at position 38. Conversely, Env acquiring the V38A mutation in a 140I background induced a significantly reduced loss of CD4+ T cells and lower single-cell death than did their baseline controls. No altered ability to induce single-cell death was observed in the other clones. Overall, primary gp41 proteins with both V38A and N140I changes showed a reduced ability to induce single cell death and deplete CD4+ T cells, despite maintaining fusion activity. The specificity of this phenotype highlights the relevance of the genetic context to the cytopathic capacity of Env and the role of ENF-resistance mutations in modulating viral pathogenicity in vivo, further supporting the hypothesis that gp41 is a critical mediator of HIV pathogenesis.

• Psychological stress is associated with high levels of IL-6 in HIV-1 infected individuals on effective combined antiretroviral treatment.

  Authors: Carmina R Fumaz, Marian Gonzalez-Garcia, Xavier Borras, Jose A Muñoz-Moreno, Nuria Perez-Alvarez, Beatriz Mothe, Christian Brander, Maria Jose Ferrer, Jordi Puig, Anuska Llano, Jordi Fernandez-Castro, Bonaventura Clotet

  Brain, behavior, and immunity. 01/2012;

  This study explores the role of psychological stress in the circulating levels of interleukin-6 (IL-6) in a group of HIV-1 infected individuals on effective cART. We developed a cross-sectional studyThis study explores the role of psychological stress in the circulating levels of interleukin-6 (IL-6) in a group of HIV-1 infected individuals on effective cART. We developed a cross-sectional study with 50 individuals with confirmed diagnosis of HIV-1 infection ⩾1 and ⩽8years, on continuous cART for >1 and <8years and with plasma viral load <50 copies/mL for at least 1year. Clinical, behavioral and psychological variables were collected to control their possible indirect contribution in the relationship between psychological stress and IL-6. Pearson correlation and univariate/multivariate logistic regressions were performed. Eighty-eight percent of the subjects were male: median (IQR) age: 39.0 (32.7-42.2), years since HIV-1 infection: 3.4 (2.1-7.0), years on cART: 2.5 (1.6-5.7), CD4 cell count: 709.0 (573.5-881.0) cell/mm(3), plasma levels of IL-6: 7.0 (0-12.2) pg/ml. A strong correlation between IL-6 and psychological stress was found (r=.81). Psychological stress (coef: 0.49; SD: 0.05), anxiety/depression (0.37; 0.08) and unhealthy diet (2.94; 1.38) were associated with higher levels of IL-6. In the multivariate model psychological stress remained strongly associated with IL-6 (R(2): 59%). In conclusion, individuals with psychological stress presented high levels of IL-6 and psychological stress was the only variable which remained strongly associated with IL-6. This strong relationship suggests evidence for a mechanism through which psychological stress might contribute to the health's impairment of HIV-infected individuals on effective cART.

• Differential prevalence of the HLA-C -35 CC genotype among viremic long term non-progressor and elite controller HIV+ individuals.

  Authors: Ester Ballana, Alba Ruiz-de Andres, Beatriz Mothe, Eva Ramirez de Arellano, Francisco Aguilar, Roger Badia, Eulalia Grau, Bonaventura Clotet, Margarita Del Val, Christian Brander, José A Esté

  Immunobiology. 01/2012;

  Susceptibility to HIV infection and disease progression are complex traits modulated by environmental and genetic factors, affecting innate and adaptive immune responses, among other cellularSusceptibility to HIV infection and disease progression are complex traits modulated by environmental and genetic factors, affecting innate and adaptive immune responses, among other cellular processes. A single nucleotide polymorphism (SNP) 35kb upstream of the HLA-C gene locus (-35C/T) was previously shown to correlate with increased HLA-C expression and improved control of HIV-1. Here, we genotyped the -35C/T SNP in 639 subjects (180 uninfected patients, 304 HIV progressors and 155 LTNP) and confirmed the association of the -35C/T variant with the LTNP phenotype. The genotype frequencies in the general population subjects did not differ significantly from those seen in HIV progressors (p-value=0.472). However, a significant higher frequency of the protective CC genotype was identified when LTNP were compared either with HIV progressors alone (p-value<0.0001) or progressors and uninfected subjects together (p-value<0.0001). When considering aviremic LTNP alone (elite controllers; viral load below 50copies/ml), the -35 CC genotype was not overrepresented compared to HIV progressors. Conversely, a significant association was found with the viremic LTNP groups (viral loads below 10,000copies/ml). These results suggest that other factors alone or in combination with the -35 CC genotype may play an important role in differentiating the elite controller status from LTNP. Combination of different genetic variants may have additive or epistatic effects determining the HIV course of infection.

• Susceptibility of Human Lymphoid Tissue Cultured ex vivo to Xenotropic Murine Leukemia Virus-Related Virus (XMRV) Infection.

  Authors: Marta Curriu, Jorge Carrillo, Marta Massanella, Elisabet Garcia, Francesc Cunyat, Ruth Peña, Peter Wienberg, Cristina Carrato, Joan Areal, Margarita Bofill, Bonaventura Clotet, Julià Blanco, Cecilia Cabrera

  PloS one. 01/2012; 7(5):e37415.

  Xenotropic murine leukemia virus-related virus (XMRV) was generated after a recombination event between two endogenous murine leukemia viruses during the production of a prostate cancer cell line.Xenotropic murine leukemia virus-related virus (XMRV) was generated after a recombination event between two endogenous murine leukemia viruses during the production of a prostate cancer cell line. Although the associations of the XMRV infection with human diseases appear unlikely, the XMRV is a retrovirus of undefined pathogenic potential, able to replicate in human cells in vitro. Since recent studies using animal models for infection have yielded conflicting results, we set out an ex vivo model for XMRV infection of human tonsillar tissue to determine whether XMRV produced by 22Rv1 cells is able to replicate in human lymphoid organs. Tonsil blocks were infected and infection kinetics and its pathogenic effects were monitored XMRV, though restricted by APOBEC, enters and integrates into the tissue cells. The infection did not result in changes of T or B-cells, immune activation, nor inflammatory chemokines. Infectious viruses could be recovered from supernatants of infected tonsils by reinfecting DERSE XMRV indicator cell line, although these supernatants could not establish a new infection in fresh tonsil culture, indicating that in our model, the viral replication is controlled by innate antiviral restriction factors. Overall, the replication-competent retrovirus XMRV, present in a high number of laboratories, is able to infect human lymphoid tissue and produce infectious viruses, even though they were unable to establish a new infection in fresh tonsillar tissue. Hereby, laboratories working with cell lines producing XMRV should have knowledge and understanding of the potential biological biohazardous risks of this virus.

• CTL responses of high functional avidity and broad variant cross-reactivity are associated with HIV control.

  Authors: Beatriz Mothe, Anuska Llano, Javier Ibarrondo, Jennifer Zamarreño, Mattia Schiaulini, Cristina Miranda, Marta Ruiz-Riol, Christoph T Berger, M José Herrero, Eduard Palou [......] David Heckerman, Florencia Pereyra, Bruce D Walker, David Weiner, Roger Paredes, Bonaventura Clotet, Barbara K Felber, George N Pavlakis, James I Mullins, Christian Brander

  PloS one. 01/2012; 7(1):e29717.

  Cytotoxic T lymphocyte (CTL) responses targeting specific HIV proteins, in particular Gag, have been associated with relative control of viral replication in vivo. However, Gag-specific CTL can alsoCytotoxic T lymphocyte (CTL) responses targeting specific HIV proteins, in particular Gag, have been associated with relative control of viral replication in vivo. However, Gag-specific CTL can also be detected in individuals who do not control the virus and it remains thus unclear how Gag-specific CTL may mediate the beneficial effects in some individuals but not in others. Here, we used a 10mer peptide set spanning HIV Gag-p24 to determine immunogen-specific T-cell responses and to assess functional properties including functional avidity and cross-reactivity in 25 HIV-1 controllers and 25 non-controllers without protective HLA class I alleles. Our data challenge the common belief that Gag-specific T cell responses dominate the virus-specific immunity exclusively in HIV-1 controllers as both groups mounted responses of comparable breadths and magnitudes against the p24 sequence. However, responses in controllers reacted to lower antigen concentrations and recognized more epitope variants than responses in non-controllers. These cross-sectional data, largely independent of particular HLA genetics and generated using direct ex-vivo samples thus identify T cell responses of high functional avidity and with broad variant reactivity as potential functional immune correlates of relative HIV control.

• Viremic HIV infected individuals with high CD4 T cells and functional envelope proteins show anti-gp41 antibodies with unique specificity and function.

  Authors: Marta Curriu, Hughes Fausther-Bovendo, María Pernas, Marta Massanella, Jorge Carrillo, Cecilia Cabrera, Cecilio López-Galíndez, Bonaventura Clotet, Patrice Debré, Vincent Vieillard, Julià Blanco

  PloS one. 01/2012; 7(2):e30330.

  CD4 T-cell decay is variable among HIV-infected individuals. In exceptional cases, CD4 T-cell counts remain stable despite high plasma viremia. HIV envelope glycoprotein (Env) properties, namelyCD4 T-cell decay is variable among HIV-infected individuals. In exceptional cases, CD4 T-cell counts remain stable despite high plasma viremia. HIV envelope glycoprotein (Env) properties, namely tropism, fusion or the ability to induce the NK ligand NKp44L, or host factors that modulate Env cytopathic mechanisms may be modified in such situation. We identified untreated HIV-infected individuals showing non-cytopathic replication (VL>10,000 copies/mL and CD4 T-cell decay<50 cells/µL/year, Viremic Non Progressors, VNP) or rapid progression (CD4 T-cells<350 cells/µL within three years post-infection, RP). We isolated full-length Env clones and analyzed their functions (tropism, fusion activity and capacity to induce NKp44L expression on CD4 cells). Anti-Env humoral responses were also analyzed. Env clones isolated from VNP or RP individuals showed no major phenotypic differences. The percentage of functional clones was similar in both groups. All clones tested were CCR5-tropic and showed comparable expression and fusogenic activity. Moreover, no differences were observed in their capacity to induce NKp44L expression on CD4 T cells from healthy donors through the 3S epitope of gp41. In contrast, anti- Env antibodies showed clear functional differences: plasma from VNPs had significantly higher capacity than RPs to block NKp44L induction by autologous viruses. Consistently, CD4 T-cells isolated from VNPs showed undetectable NKp44L expression and specific antibodies against a variable region flanking the highly conserved 3S epitope were identified in plasma samples from these patients. Conversely, despite continuous antigen stimulation, VNPs were unable to mount a broad neutralizing response against HIV. Env functions (fusion and induction of NKp44L) were similar in viremic patients with slow or rapid progression to AIDS. However, differences in humoral responses against gp41 epitopes nearby 3S sequence may contribute to the lack of CD4 T cell decay in VNPs by blocking the induction of NKp44L by gp41.

• Deciphering the interleukin 28B variants that better predict response to pegylated interferon-α and ribavirin therapy in HCV/HIV-1 coinfected patients.

  Authors: Montserrat de Castellarnau, Ester Aparicio, Mariona Parera, Sandra Franco, Cristina Tural, Bonaventura Clotet, Miguel Angel Martínez

  PloS one. 01/2012; 7(2):e31016.

  Previous works have documented the contribution of different IL28B-associated SNPs to spontaneous HCV clearance. This study investigated the effect of different interleukin (IL) 28B genetic variantsPrevious works have documented the contribution of different IL28B-associated SNPs to spontaneous HCV clearance. This study investigated the effect of different interleukin (IL) 28B genetic variants on interferon (IFN)-based therapy response. We genotyped eight IL28B single-nucleotide polymorphisms (SNPs) in a cohort of 197 hepatitis C virus (HCV)/human immunodeficiency virus type 1 (HIV-1) coinfected patients from our clinic unit who received combined pegylated (peg)-IFN-α and ribavirin (RBV) therapy. This analysis included the two strongest tag predictors for HCV clearance, rs8099917 and rs12979860, and four causal variants (rs4803219, rs28416813, rs8103142, and rs4803217) located in the IL28B promoter, coding, and 3'-untranslated regions. Haplotypes carrying the major alleles at IL28B SNPs were highly associated with sustained virological responses (SVRs) after treatment with peg-IFN-α and RBV [odds ratio (OR) = 2.5, 95% confidence interval (CI) = 1.6-4.0, 4.0×10(-5)]. Three causal SNP genotypes (rs28416813, rs8103142, and rs4803217) displayed the highest association with SVRs (OR = 3.7, 95% CI = 2.0-6.7, p = 1.3×10(-5)). All four causal variants were in high linkage disequilibrium, both among themselves (r(2)≥0.94) and with the rs12979860 variant (r(2)≥0.92). In contrast, rs8099917 was in low linkage disequilibrium with the four causal variants (r(2)≤0.45) and with the rs12979860 variant (r(2) = 0.45). These results demonstrate that rs12979860, compared to rs8099917, may be a better predictor of response to the peg-IFN/RBV treatment among HCV/HIV-1 coinfected patients. Moreover, causal IL28B variants are strongly associated with treatment SVRs.

• In-Depth Characterization of Viral Isolates from Plasma and Cells Compared with Plasma Circulating Quasispecies in Early HIV-1 Infection.

  Authors: Judith Dalmau, Francisco M Codoñer, Itziar Erkizia, Maria Pino, Christian Pou, Roger Paredes, Bonaventura Clotet, Javier Martinez-Picado, Julia G Prado

  PloS one. 01/2012; 7(2):e32714.

  The use of in vitro models to unravel the phenotypic characteristics of circulating viral variants is key to understanding HIV-1 pathogenesis but limited by the availability of primary viral isolatesThe use of in vitro models to unravel the phenotypic characteristics of circulating viral variants is key to understanding HIV-1 pathogenesis but limited by the availability of primary viral isolates from biological samples. However, overall in vivo genetic variability of HIV-1 within a subject may not be reflected in the viable viral population obtained after isolation. Although several studies have tried to determine whether viral populations expanded in vitro are representative of in vivo findings, the answer remains unclear due to the reduced number of clonal sequences analyzed or samples compared. In order to overcome previous experimental limitations, here we applied Deep Pyrosequencing (DPS) technology in combination with phenotypic experiments to analyze and compare with unprecedented detail the composition of viral isolates and in vivo quasispecies. We amplified by DPS HIV-1 genomic regions covering gag, protease, integrase and env-V3 to characterize paired isolates from plasma and peripheral blood mononuclear cells and compare them with total plasma viral RNA in four recently HIV-1 infected subjects. Our study demonstrated the presence of unique haplotypes scattered between sample types with conservation of major variants. In addition, no differences in intra- and inter-population encoded protein variability were found between the different types of isolates or when these were compared to plasma viral RNA within subjects. Additionally, in vitro experiments demonstrated phenotypic similarities in terms of replicative capacity and co-receptor usage between viral isolates and plasma viral RNA. This study is the first in-depth comparison and characterization of viral isolates from different sources and plasma circulating quasispecies using DPS in recently HIV-1 infected subjects. Our data supports the use of primary isolates regardless of their plasma or cellular origin to define genetic variability and biological traits of circulating HIV-1 quasispecies.