Sensitive quantification of the HIV-1 reservoir in gut-associated lymphoid tissue

Biòpsia; VIH-1; Teixit limfoideBiopsia; VIH-1; Tejido linfoideBiopsy; HIV-1; Lymphoid tissueBackground The implementation of successful strategies to achieve an HIV cure has become a priority in HIV research. However, the current location and size of HIV reservoirs is still unknown since there are limited tools to evaluate HIV latency in viral sanctuaries such as gut-associated lymphoid tissue (GALT). As reported in the so called “Boston Patients”, despite undetectable levels of proviral HIV-1 DNA in blood and GALT, viral rebound happens in just few months after ART interruption. This fact might imply that current methods are not sensitive enough to detect residual reservoirs. Showing that, it is imperative to improve the detection and quantification of HIV-1 reservoir in tissue samples. Herein, we propose a novel non-enzymatic protocol for purification of Lamina Propria Leukocytes (LPL) from gut biopsies combined to viral HIV DNA (vDNA) quantification by droplet digital PCR (ddPCR) to improve the sensitivity and accuracy of viral reservoir measurements (LPL-vDNA assay). Methods Endoscopic ileum biopsies were sampled from 12 HIV-1-infected cART-suppressed subjects. We performed a DTT/EDTA-based treatment for epithelial layer removal followed by non-enzymatic disruption of the tissue to obtain lamina propria cell suspension (LP). CD45+ cells were subsequently purified by flow sorting and vDNA was determined by ddPCR. Results vDNA quantification levels were significantly higher in purified LPLs (CD45+) than in bulk LPs (p<0.01). The levels of vDNA were higher in ileum samples than in concurrent PBMC from the same individuals (p = 0.002). As a result of the increased sensitivity of this purification method, the Poisson 95% confidence intervals of the vDNA quantification data from LPLs were narrower than that from bulk LPs. Of note, vDNA was unambiguously quantified above the detection limit in 100% of LPL samples, while only in 58% of bulk LPs. Conclusion We propose an innovative combined protocol for a more sensitive detection of the HIV reservoir in gut-associated viral sanctuaries, which might be used to evaluate any proposed eradication strategy.This substudy was supported by ViiV and the American Foundation for AIDS Research (amfAR) (ARCHE). IrsiCaixa was supported by the CERCA programme from Generalitat de Catalunya. MS was supported by the post-doctoral training scholarship (Juan de la Cierva) of the Spanish Economy and Competitiveness Ministry (FPDI-2013-17134). SM-L received a fellowship from the Agència de Gestió d’Ajuts Universitaris i de Recerca (2013FI_B 00275). CG was partially supported by the pre-doctoral fellowship of the Spanish Education, Culture and Sport Ministry (FPU15/03698). The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication

Long-term effectiveness of first-line antiretroviral theraphy in a cohort of HIV-1 infected patients

Eligibility criteria might explain differences in viral response to combined antiretroviral treatment (cART) between clinical trials and routine care setting. Prospective analysis including HIV-1 infected patients starting cART between January 2004 and December 2009, at Hospital Universitari Vall d'Hebron. Effectiveness evaluated as time to treatment failure (TF), defined as virologic failure, loss to follow-up, death or treatment discontinuation whatever the reason other than switching. Effectiveness month 12, 24 and 36 was 82.9%, 78.5% and 76%, respectively. 57 (24.6%) patients presented TF, mainly due to intolerance or toxicity. Higher risk in patients starting before 2006 and those with protease inhibitor based regimen.Els criteris d'inclusió podrien explicar les diferències en quant a resposta viral al tractament antiretrovial (cART) entre assajos clínics i pràctica clínica. Estudi prospectiu amb pacients VIH-1 que iniciaren cART entre gener 2004 i desembre 2009 a l'Hospital Universitari Vall d'Hebron. L'efectivitat s'avaluà com temps fins fracàs del tractament (TF), definit com fracàs virològic, pèrdua de seguiment, mort o canvi de pauta, excepte per simplificació. L'efectivitat mes 12, 24 i 36 fou 82.9%, 78.5% i 76%, respectivament. 57 (24.6%) pacients presentaren TF, principalment per intolerància o toxicitat. Risc més elevat en els iniciats abans del 2006 i en règims basats en inhibidors de proteasa

Low nadir CD4+ T-cell counts predict gut dysbiosis in HIV-1 infection

Human immunodeficiency virus (HIV)-1 infection causes severe gut and systemic immune damage, but its effects on the gut microbiome remain unclear. Previous shotgun metagenomic studies in HIV-negative subjects linked low-microbial gene counts (LGC) to gut dysbiosis in diseases featuring intestinal inflammation. Using a similar approach in 156 subjects with different HIV-1 phenotypes, we found a strong, independent, dose-effect association between nadir CD4+ T-cell counts and LGC. As in other diseases involving intestinal inflammation, the gut microbiomes of subjects with LGC were enriched in gram-negative Bacteroides, acetogenic bacteria and Proteobacteria, which are able to metabolize reactive oxygen and nitrogen species; and were depleted in oxygen-sensitive methanogenic archaea and sulfate-reducing bacteria. Interestingly, subjects with LGC also showed increased butyrate levels in direct fecal measurements, consistent with enrichment in Roseburia intestinalis despite reductions in other butyrate producers. The microbiomes of subjects with LGC were also enriched in bacterial virulence factors, as well as in genes associated with beta-lactam, lincosamide, tetracycline, and macrolide resistance. Thus, low nadir CD4+ T-cell counts, rather than HIV-1 serostatus per se, predict the presence of gut dysbiosis in HIV-1 infected subjects. Such dysbiosis does not display obvious HIV-specific features; instead, it shares many similarities with other diseases featuring gut inflammation.Fundació Glòria SolerFundació Catalunya-La PedreraGala SIDA 2015-2016Nit per la Recerca a la Catalunya Central 2015 editionPeople in Red-Barcelona 2016 editionRED de SIDA RD16/0025/0041ISCIIIEuropean Regional Develpment Fund (ERDF)Agencia de Gestio d´Ajuts Universitaris i de Recerca (AGAUR)Secretaria d´Universitats i Recerca del Departament d´Economia i Coneixement de la Generalitat de CatalunyaMinisterio de Economia y Competitividad. EspañaUniversidad de Whashingto

Determinants and prognostic value of peak oxygen uptake in the current era of heart failure disease modifying therapy

Abstrac

Tratamiento de la hepatitis crónica C en pacientes coinfectados por VIH

Consultable des del TDXTítol obtingut de la portada digitalitzadaEn esta tesis, se recogen los resultados de un estudio aleatorizado y comparativo llevado a cabo en nuestro centro, en el que se demuestra que la combinación de peginterferon alfa-2b y ribavirina es igual de segura y más eficaz que el interferón convencional y la ribavirina para el tratamiento del VHC en pacientes coinfectados por VIH. Es preciso destacar la importancia de la monitorización durante el tratamiento las manifestaciones clínicas secundarias a toxicidad mitocondrial, asociada con la co-administración de ribavirina y determinados fármacos antiretrovirales, sobre todo con didanosina, que debe ser evitada. Finalmente, destaca la importancia práctica de la monitorización de la dinámica de respuesta durante las primeras semanas de tratamiento. Así, la respuesta virológica observada en las semanas 4ª, 8ª y 12ª nos permite detectar a los pacientes sin posibilidad respuesta virológica a los que podremos suspender precozmente el tratamiento. Este hecho, aparte del ahorro económico que supone, tiene especial relevancia en los pacientes coinfectados por el mayor riesgo de toxicidad asociado al tratamiento concomitante de ambas infecciones. Además, nuestros resultados cuestionan la validez de un reciente documento de consenso en el que se aconseja tratar a todos los pacientes coinfectados por VIH durante 48 semanas con independencia del genotipo del VHC. En este estudio, los pacientes con genotipo 3 que muestran una respuesta virológica rápida (ARN VHC indetectable en la semana 4ª de tratamiento) pueden ser tratados durante 24 semanas con un riesgo de recidiva muy bajo. Aun considerando las limitaciones de este estudio como el limitado número de pacientes con genotipo 3 incluido y el análisis post-hoc de los datos, estos resultados sugieren que la monitorización de la respuesta virológica en la 4ª semana de tratamiento puede ser utilizada como guía para individualizar la duración del tratamiento en este subgrupo de pacientes, que representa el 25% del total de pacientes tratados. Otro aspecto analizado es la utilidad de la monitorización de la concentración de ribavirina en suero como guía para ajustar e individualizar precozmente la dosis de ribavirina, con la finalidad de disminuir la toxicidad y aumentar su eficacia. Entre los resultados obtenidos, destaca que la concentración valle sérica de ribavirina depende de la dosis administrada ajustada al peso del paciente, y que la co-administración de tenofovir se asoció con concentraciones más elevadas de ribavirina. A diferencia de la mayoría de los datos preliminares, publicados recientemente en otros estudios, observamos una correlación inversa entre la concentración de ribavirina en suero y la respuesta virológica en la semana 4ª de tratamiento. Estos datos concuerdan con los resultados publicados recientemente por Dahari et al (J Hepatol 2007; 47:1 23-30) y sugieren que la incorporación intracelular de ribavirina durante las primeras semanas de tratamiento pude mejorar la dinámica de respuesta del VHC al tratamiento. Sin embargo, en nuestro estudio, la concentración de ribavirina en suero mostró escasa capacidad para discernir entre los pacientes con y sin respuesta virológica en las semanas 4ª y 12ª de tratamiento. Además, correlación entre la concentración de ribavirina y el descenso de hemoglobina fue débil. Por tanto, los datos de este estudio no apoyan la utilidad de la monitorización de ribavirina en suero en la optimización del tratamiento del VHC en pacientes coinfectados por VIH. No obstante, son necesarios nuevos estudios para verificar la hipótesis de la incorporación intracelular de ribavirina y la dinámica de respuesta precoz, y estudios que aborden la potencial interacción entre el tenofovir y la ribavirina. Finalmente, hemos evaluado la eficacia del tratamiento con peginterferon alfa-2b y ribavirina en pacientes con historia de fracaso virológico previo al tratamiento con interferón convencional y ribavirina. Los resultados obtenidos deben merecen una interpretación cautelosa por el limitado número de pacientes incluidos. El 43% (9/21) de los pacientes obtuvo respuesta virológica al final de las 48 semanas de tratamiento, a pesar de las características pre-tratamiento adversas (genotipo 1/4, 76%; cirrosis 42%) y de que hasta el 71% de los pacientes precisó disminuir la dosis de peginterferon por efectos adversos. No obstante, se produjo un elevado porcentaje de recidivas (55%) por lo que sólo 4 pacientes (19%) obtuvo una respuesta virológica mantenida. Estos datos sugieren que son necesarias estrategias de tratamiento individualizadas que refuercen el cumplimiento terapéutico o bien pautas de tratamiento más prolongadas para mejorar la eficacia en este subgrupo de tratamiento muy difícil

Low nadir CD4+ T-cell counts predict gut dysbiosis in HIV-1 infection

Human immunodeficiency virus (HIV)-1 infection causes severe gut and systemic immune damage, but its effects on the gut microbiome remain unclear. Previous shotgun metagenomic studies in HIV-negative subjects linked low-microbial gene counts (LGC) to gut dysbiosis in diseases featuring intestinal inflammation. Using a similar approach in 156 subjects with different HIV-1 phenotypes, we found a strong, independent, dose-effect association between nadir CD4+ T-cell counts and LGC. As in other diseases involving intestinal inflammation, the gut microbiomes of subjects with LGC were enriched in gram-negative Bacteroides, acetogenic bacteria and Proteobacteria, which are able to metabolize reactive oxygen and nitrogen species; and were depleted in oxygen-sensitive methanogenic archaea and sulfate-reducing bacteria. Interestingly, subjects with LGC also showed increased butyrate levels in direct fecal measurements, consistent with enrichment in Roseburia intestinalis despite reductions in other butyrate producers. The microbiomes of subjects with LGC were also enriched in bacterial virulence factors, as well as in genes associated with beta-lactam, lincosamide, tetracycline, and macrolide resistance. Thus, low nadir CD4+ T-cell counts, rather than HIV-1 serostatus per se, predict the presence of gut dysbiosis in HIV-1 infected subjects. Such dysbiosis does not display obvious HIV-specific features; instead, it shares many similarities with other diseases featuring gut inflammation

Long-term safety and efficacy of emtricitabine and tenofovir alafenamide vs emtricitabine and tenofovir disoproxil fumarate for HIV-1 pre-exposure prophylaxis: week 96 results from a randomised, double-blind, placebo-controlled, phase 3 trial

In DISCOVER, a multinational, randomised controlled trial, emtricitabine and tenofovir alafenamide compared with emtricitabine and tenofovir disoproxil fumarate showed non-inferior efficacy for HIV prevention and improved bone mineral density and renal safety biomarkers at week 48. We report outcomes analysed after all participants had completed 96 weeks of follow-up. This study is an ongoing, randomised, double-blind, multicentre, active-controlled, phase 3, non-inferiority trial done at 94 community, public health, and hospital-associated clinics located in Europe and North America. Adult cisgender men and transgender women who have sex with men, both with a high risk of acquiring HIV as determined by self-reported sexual behaviour or recent sexually transmitted infections, were randomly assigned (1:1) to receive either emtricitabine and tenofovir alafenamide (200/25 mg) tablets daily, with matched placebo tablets (emtricitabine and tenofovir alafenamide group), or emtricitabine and tenofovir disoproxil fumarate (200/300 mg) tablets daily, with matched placebo tablets (emtricitabine and tenofovir disoproxil fumarate group). The primary efficacy outcome was incident HIV infection. Incidence of HIV-1 infection per 100 person-years was assessed when the last participant had completed 96 weeks of follow-up. This trial is registered with ClinicalTrials.gov, number NCT02842086. Between Sept 13, 2016, and June 30, 2017, 5387 participants were randomly assigned to receive emtricitabine and tenofovir alafenamide (n=2694) or emtricitabine and tenofovir disoproxil fumarate (n=2693), contributing 10 081 person-years of follow-up. At 96 weeks of follow-up, there were eight HIV infections in participants who had received emtricitabine and tenofovir alafenamide (0·16 infections per 100 person-years [95% CI 0·07–0·31]) and 15 in participants who had received emtricitabine and tenofovir disoproxil fumarate (0·30 infections per 100 person-years [0·17–0·49]). Emtricitabine and tenofovir alafenamide maintained its non-inferiority to emtricitabine and tenofovir disoproxil fumarate for HIV prevention (IRR 0·54 [95% CI 0·23–1·26]). Approximately 78–82% of participants reported taking study medication more than 95% of the time across all study visits. Rates of sexually transmitted infections remained high and similar across groups (21 cases per 100 person-years for rectal gonorrhoea and 28 cases per 100 person-years for rectal chlamydia). Emtricitabine and tenofovir alafenamide continued to show superiority over emtricitabine and tenofovir disoproxil fumarate in all but one of the six prespecified bone mineral density and renal biomarkers. There was more weight gain among participants who had received emtricitabine and tenofovir alafenamide (median weight gain 1·7 kg vs 0·5 kg, p<0·0001). Emtricitabine and tenofovir alafenamide is safe and effective for longer-term pre-exposure prophylaxis in cisgender men and transgender women who have sex with men. Gilead Sciences