Uso clínico de Maraviroc (MRV) en pacientes con infección por VIH-1: diseño de una nueva estrategia para la determinación de la sensibilidad clínica a MRV mediante una exposición a corto plazo al fármaco. Evolución inmunovirológica y seguridad a largo plazo de un régimen antirretroviral conteniendo MRV

Se trata de una tesis doctoral presentada en la modalidad compendio de publicaciones. En ella, se presenta una línea de trabajo de investigación mediante la presentación sucesiva de tres artículos ya publicados en diferentes revistas de impacto en el ámbito de las enfermedades infecciosas. Los trabajos presentados en esta tesis nacen de una necesidad clínica patente en nuestra asistencia a los pacientes con infección por VIH, ya que el uso clínico de maraviroc, un antagonista del correceptor de quimiocinas CCR5, se veía dificultado por la necesidad de conocer el tropismo vírico y los métodos disponibles para ello eran caros, complejos y con problemas de interpretación. Por ello, el primer trabajo de la presente tesis fue diseñar una herramienta clínica más sencilla rápida y barata que los métodos disponibles hasta entonces; además, esta herramienta clasificaba al paciente en sensible al fámaco o no, más allá de un resultado categórico de tropismo. Este objetivo se abordó en el primer artículo publicado en "Journal of Antimicrobial Chemotherapy" en 2009. Posteriormente, realizamos un análisis de comparación entre nuestra herramienta clínica y el método fenotípico estándar de oro para determinar el tropismo vírico, con el onjetivo de alertar sobre las implicaciones clínicas que podrían tener tasas de discrepancia entre ambos métodos superiores al 15%. Este segundo objetivo se en el segundo artículo publicado en "Antiviral Research" en 2011. Por último, nos interesaba conocer cuál era la eficacia y seguridad de maraviroc asociado a diferentes antirretrovirales en nuestra cohorte de pacientes, masivamente coinfectada por virus de la hepatitis C, y en diferentes escenarios clínicos. Además, en la mayoría de estos pacientes se decidió iniciar tratamiento con maraviroc a través de la herramienta clínica previamente diseñada. Este tercer objetivo se abordó en el último trabajo publicado en "Current HIV Research" en 2010. Los resultados obtenidos en estos tres trabajos permiten extraer las siguientes conclusiones: 1. Una herramienta clínica que permite determinar la sensibilidad clínica a maraviroc es un método sencillo y eficaz para considerar a un paciente candidato a recibir tratamiento con maraviroc. 2. Se observan tasas de discrepancia entre esta herramienta clínica y los métodos fenotípicos superiores al 15%, lo que puede tener importantes repercusiones clínicas. 3. Un régimen antirretroviral con maraviroc es eficaz y seguro en diferentes escenarios clínicos, asociado a diferentes antirretrovirales y en una población con elevada tasa de coinfección por virus de la hepatitis C.Premio Extraordinario de Doctorado U

altered expression of cD300a inhibitory receptor on cD4+ T cells From human immunodeficiency Virus-1-infected Patients: association With Disease Progression Markers.

The ability of the CD300a inhibitory receptor to modulate immune cell functions and its involvement in the pathogenesis of many diseases has aroused a great interest in this molecule. Within human CD4+ T lymphocytes from healthy donors, the inhibitory receptor CD300a is differentially expressed among different T helper subsets. However, there are no data about the expression and regulation of CD300a receptor on CD4+ T cells from human immunodeficiency virus (HIV)-1-infected patients. The objective of this study was to investigate the expression of CD300a on CD4+ T cells from HIV-infected patients on suppressive combined antiretroviral therapy (cART) and cART naïve patients. Our results have demonstrated that the expression levels of this inhibitory receptor were higher on CD4+ T cells from HIV-1 infected subjects compared with healthy donors, and that cART did not reverse the altered expression of CD300a receptor in these patients. We have observed an increase of CD300a expression on both PD1+CD4+ and CD38+CD4+ T cells from HIV-1 infected people. Interestingly, a triple positive (CD300a+PD1+CD38+) subset was expanded in naïve HIV-1 infected patients, while it was very rare in healthy donors and patients on cART. Finally, we found a negative correlation of CD300a expression on CD4+ T lymphocytes and some markers associated with HIV-1 disease progression. Thus, our results show that HIV-1 infection has an impact in the regulation of CD300a inhibitory receptor expression levels, and further studies will shed light into the role of this cell surface receptor in the pathogenesis of HIV infection

HIV-Infected Subjects With Poor CD4 T-Cell Recovery Despite Effective Therapy Express High Levels of OX40 and α4β7 on CD4 T-Cells Prior Therapy Initiation

Background HIV-infected subjects with suboptimal CD4 restoration despite suppressive combined antiretroviral treatment (cART) (immunodiscordant subjects) have been classically characterized after a variable period of time under cART. Recently, we have reported that an increased frequency of proliferating CD4 T-cells in these subjects is already present before the cART onset. The potential contribution of peripheral compensatory homeostatic proliferation (HP) is yet unknown. We aimed to analyze the expression of HP-related cellular markers on CD4 T-cells of immunodiscordant subjects before cART. Go to: Methods We analyzed the expression of OX40 and α4β7 on peripheral CD4 T-cells from immunodiscordant and control subjects (n = 21 each group) before cART initiation, and also on available follow-up samples (after 24 month of suppressive cART). Additionally, we tested the expression of these markers in an in vitro system for the study of human HP processes. Go to: Results Immunodiscordant subjects showed increased levels of OX40 and α4β7 on CD4 T-cells before cART initiation. While the cART tended to reduce these levels, immunodiscordant subjects still maintained comparatively higher levels of OX40 and α4β7 after 24 months under suppressive cART. These HP-related markers were upregulated in vitro during the human HP, especially during the fast HP. Go to: Conclusion Our results are compatible with exacerbated HP processes in immunodiscordant subjects, already before the cART onset.Fondo de Investigación Sanitaria FIS PI14/01693 PI13/0796 PI16/0503Fondos Europeos para el Desarrollo Regional (FEDER) CTS2593Junta de Andalucía. Consejería de Economía, Innovación, Ciencia y Empleo CTS2593AGAUR 2017SGR948GILEAD GLD14/293The Spanish AIDS Research Network of Excellence RD12/0017/0029 RD16/0025/0019 RD16/0025/0006Junta de Andalucía. Consejería de Salud y Bienestar Social C-0013-201

A Lower Baseline CD4/CD8 T-Cell Ratio Is Independently Associated with Immunodiscordant Response to Antiretroviral Therapy in HIV-Infected Subjects

We explored if baseline CD4/CD8 T-cell ratio is associated with immunodiscordant response to antiretroviral therapy in HIV-infected subjects. Comparing immunodiscordant and immunoconcordant subjects matched by pretreatment CD4 counts, we observed a lower pretreatment CD4/CD8 T-cell ratio in immunodiscordant subjects. Furthermore, pretreatment CD4/CD8 T-cell ratio, but not CD4 counts, correlated with the main immunological alterations observed in immunodiscordants, including increased regulatory T-cell (Treg) frequency and T-cell turnover-related markers. Then, in a larger cohort, only baseline CD4/CD8 T-cell ratio was independently associated with immunodiscordance, after adjusting by the viral CXCR4-tropic HIV variants. Our results suggest that the CD4/CD8 T-cell ratio could be an accurate biomarker of the subjacent immunological damage triggering immunodiscordance.The HIV BioBank, integrated in the Spanish AIDS Research Network, is supported by Instituto de Salud Carlos III, Spanish Health Ministry (grants RD06/0006/0035 and RD12/0017/0037), as part of the Plan Nacional R + D + I and cofinanced by ISCIII-Subdirección General de Evaluación y el Fondo Europeo de Desarrollo Regional (FEDER) and Fundación para la investigación y prevención del SIDA en España (FIPSE). The RIS Cohort (CoRIS) is funded by the Instituto de Salud Carlos III through the Red Temática de Investigación Cooperativa en SIDA by the RD12/0017/0018 project, as part of the Plan Nacional R + D + I, and cofinanced by ISCIII-Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (FEDER). The project number is RIS_EPICLIN 20/2015. This work was supported by grants from the Fondo de Investigación Sanitaria, ISCIII (FIS; PI14/01693), and the Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía (Proyecto de Investigación de Excelencia; CTS2593), and cofunded by Fondos Europeos para el Desarrollo Regional (FEDER). I. Rosado-Sánchez was supported by the Spanish AIDS Research Network of Excellence (RIS; RD12/0017/0029). Y. M. Pacheco was supported by the Fondo de Investigación Sanitaria through the Miguel Servet program (CPII13/00037) and by the Servicio Andaluz de Salud through the Nicolás Monardes program (C-0010/13).Peer reviewe

Virological Response after Short-Term CCR5 Antagonist Exposure in HIV-Infected Patients: Frequency of Subjects with Virological Response and Associated Factors▿

The virological response after an 8-day maraviroc monotherapy has been proposed to be an alternative method to determine whether an CCR5 antagonist should be prescribed to HIV-infected patients. The frequency of patients eligible for a combined antiretroviral therapy which includes maraviroc on the basis of the result of this clinical test is not well-known at the moment. In the same way, clinical and immunovirological factors associated with the virological response after antagonist exposure need to be determined. Ninety consecutive HIV-infected patients were exposed to an 8-day maraviroc monotherapy. The virological response was considered positive if either a reduction of ≥1-log10 HIV RNA copies/ml or an undetectable viral load (<40 HIV RNA copies/ml) was achieved. CXCR4- and CCR5-tropic virus levels were determined by using patients' viral isolates and multiple rounds of infection of indicator cell lines (U87-CXCR4 and U87-CCR5). The frequency of patients with a positive virological response was 72.2% (94.7% and 66.2% for treatment-naïve and pretreated patients, respectively). The positive response rates dramatically decreased in patients with lower CD4+ T-cell counts. The CXCR4-tropic virus level was the only variable independently associated with the virological response after short-term maraviroc exposure. Lower CD4+ T-cell strata were associated with higher CXCR4-tropic virus levels. These results support the suggestion that CCR5 antagonists should be an early treatment option before the expansion of CXCR4-tropic strains

Patients on a combined antiretroviral therapy after maraviroc clinical test show no immunovirological impairment

The maraviroc clinical test (MCT) is a clinical approach to establish the indication of maraviroc treatment. In this study, we analysed the long-term outcome of patients receiving a combined antiretroviral therapy (cART) selected according to MCT results. Ninety-two consecutive HIV-infected patients underwent MCT. A virological response (<40 HIV-RNA copies/ml after 24. weeks) was observed in 76/92 patients (82.6%). These patients (n= 76) were included in a time to treatment failure analysis; after a mean follow-up period of 88. weeks, treatment failure was confirmed in 14 patients (18.4%). Tropism switch during MCT was observed in 3/35 patients (8.6%); these patients experienced excellent long-term outcome on cART. In conclusion, MCT should be considered as an additional method before CCR5-antagonists prescription. © 2012 Elsevier B.V.This study was supported by ViiV Healthcare (Project Number WS843473), Redes Telemáticas de Investigación Cooperativa en Salud (RETICS; 2006, Red de SIDA RD 06/0006/0021 and RD 06/0006/035, 2007–2010) and a grant from Fondo de Investigaciones Sanitarias PS 09/01595. E. R.-M., S.F.-M. and Y.M.P. received Grants from Fondo de Investigaciones Sanitarias (CP 08/00172, FIS CD 10/00382 and CP 07/00240). We want to particularly acknowledge the HIV BioBank integrated in the Spanish AIDS Research Network, which is supported by Instituto de Salud Carlos III, the Spanish Health Ministry and Fundación para la investigación y prevención del SIDA en España (FIPSE), and collaborating centres for the generous gifts of clinical samples used in this work.Peer Reviewe

HIV-1 tropism evolution after short-term maraviroc monotherapy in HIV-1-infected patients

We analyzed the evolution of viral tropism after 8 days of maraviroc monotherapy, i.e., we used the maraviroc clinical test (MCT), in 21 patients with and 14 without virological response to the drug (MCT + and MCT - patients, respectively). No increases in CXCR4 inferred viral loads (X4IVLs) were observed in MCT + patients, while X4IVLs increased only in MCT - patients, with X4IVLs of >2 log 10 HIV RNA copies/ml. These results shed light on the evolution of viral tropism under a CCR5 antagonist in vivo. Copyright © 2012, American Society for Microbiology. All Rights Reserved.This work was supported by Pfizer (project no. WS843473), Redes Telemáticas de Investigación Cooperativa en Salud (RETICS; 2006, RED RIS RD06/0006/0021, 2007-2010; RD06/0006/0035 and RD06/0006/1004), and Fondo de Investigaciones Sanitarias (INTRASALUD RD09/0076/00103, FIS 09/01595 and 10/2635). A.G.-S. has a grant from Fundación FISEVI. E.R.-M. and S.F.-M. have grants from Fondo de Investigaciones Sanitarias (CP08/00172 and CD10/00382, respectively).Peer Reviewe

Validation of the HIV Tropism Test TROCAI Using the Virological Response to a Short-Term Maraviroc Monotherapy Exposure

TROCAI is a phenotypic tropism test developed using the virological response to a short-term exposure to maraviroc monotherapy (Maraviroc Clinical Test [MCT]). It was found that with TROCAI, a cutoff of <0.5% of dual/mixed viruses was needed to predict R5 HIV tropism. Here, we have validated TROCAI, using this cutoff, in a new cohort of 42 patients, finding a very high concordance between TROCAI and MCT (98%), and a good concordance (71 to 87%) with other genotypic/phenotypic methods.This work was supported by Redes Telemáticas de Investigación Cooperativa en Salud (RETICS; 2006, Red de SIDA RD06/0006/0021, 2007–2010 and RD16/0025/0020) and Pfizer/ViiV Healthcare (grants WS843473 and WS2425049). A.G.-S. is supported by Instituto de Salud Carlos III (CD14/00320). E.R.-M. has a grant from Fondo de Investigaciones Sanitarias (CD014/0025).Peer reviewe

Effect of maraviroc on HIV disease progression-related biomarkers

The potential effect of blocking the CCR5 receptor on HIV disease progression biomarkers is not well understood. We showed that an 8-day maraviroc (MVC) monotherapy clinical test (MCT) can be used in selecting patients to receive MVC-containing combined antiretroviral therapy (cART). Using this MCT model, we assessed the effect of MVC on several HIV disease progression biomarkers during the MCT (MVC-specific effect) and following short-term (12-week) cART. We compared 45 patients on MVC monotherapy with a control group of 25 patients on MVC-sparing cART. We found that MVC did not modify any biomarkers in patients that had no virological response after the MCT. MVC-specific effects in patients with virological responses included increased CD8 + T-cell activation and senescence levels, preservation of an increase in soluble CD14 (sCD14), and a decrease in D dimer levels. After 12 weeks, MVC-containing cART increased CD8 + T-cell counts and preserved CD4 + T-cell senescence levels compared with MVC-sparing cART. Moreover, there was a decrease in sCD14 levels in patients that received MVC-containing cART. In conclusion, effects compatible with CD8 + T-cell redistribution in peripheral blood were observed after MVC therapy. However, MVC was associated with a favorable profile in HIV disease progression biomarkers only in patients with a virological response. These results support a potential clinical benefit of a therapy which includes MVC in HIV-infected patients. Copyright © 2012, American Society for Microbiology. All Rights Reserved.This work was supported by Pfizer/ViiV Healthcare (grant number WS843473) and by Redes Temáticas de Investigación en SIDA (ISCIII RETIC RD06/0006/0021), Fondo de Investigación Sanitaria (grant PS09/01595), and the Fondos Europeos para el Desarrollo Regional (FEDER). S.F.M. and E.R.M. have grants from the Fondo de Investigaciones Sanitarias (CD10/00382 and CP08/00172, respectively). Manuel Leal has a grant from Pfizer.Peer Reviewe