Intensification of Antiretroviral Therapy with a CCR5 Antagonist in Patients with Chronic HIV-1 Infection: Effect on T Cells Latently Infected

Objective: The primary objective was to assess the effect of MVC intensification on latently infected CD4+ T cells in chronically HIV-1-infected patients receiving antiretroviral therapy. Methods: We performed an open-label pilot phase II clinical trial involving chronically HIV-1-infected patients receiving stable antiretroviral therapy whose regimen was intensified with 48 weeks of maraviroc therapy. We analyzed the latent reservoir, the residual viremia and episomal 2LTR DNA to examine the relationship between these measures and the HIV-1 latent reservoir, immune activation, lymphocyte subsets (including effector and central memory T cells), and markers associated with bacterial translocation. Results: Overall a non significant reduction in the size of the latent reservoir was found (p = 0.068). A mean reduction of 1.82 IUPM was observed in 4 patients with detectable latent reservoir at baseline after 48 weeks of intensification. No effect on plasma residual viremia was observed. Unexpectedly, all the patients had detectable 2LTR DNA circles at week 24, while none of them showed those circles at the end of the study. No changes were detected in CD4+ or CD8+ counts, although a significant decrease was found in the proportion of HLA-DR+/CD38+ CD4+ and CD8+ T-cells. LPS and sCD14 levels increased. Conclusions: Intensification with MVC was associated with a trend to a decrease in the size of the latent HIV-1 reservoir in memory T cells. No impact on residual viremia was detected. Additional studies with larger samples are needed to confirm the results

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Deep-Sequencing Analysis of the Dynamics of HIV-1 Quasiespecies in Naive Patients during a Short Exposure to Maraviroc

In this study, we have characterized quasispecies dynamics and the evolution of viral tropism in naive HIV-1-infected patients treated with a short course of maraviroc monotherapy (ClinicalTrials.gov registration no. NCT01060618) independently of the tropism of the infecting virus. We randomly selected 20 patients infected with viruses displaying different basal tropisms-10 carrying R5 and 10 carrying dual/mixed X4 (DM/X4) viruses-at recruitment as determined by phenotypic assay (Trofile). Evolution of viral quasiespecies at the end of treatment was determined by ultradeep sequencing of the V3 region using a 454 Life Sciences Platform and geno2pheno (g2p) algorithm for viral tropism prediction. The false-positive rate (FPR) that defines the probability of classifying an R5 virus falsely as X4 was set at 10%. X4-specific HIV-1 viral load (VL) was calculated from sequences with an FPR of 1-log10 copies/ml reduction in VL was detected in 70% of patients independently of the basal tropism of the infecting virus. Viral tropism remained stable, and nonsignificant differences in FPR values before and after treatment were found for the majority of patients in both tropism groups. Only three patients (one with R5 and two with DM/X4 viruses) showed an increased (>1 log) X4 VL, and one patient harboring a DM/X4-tropic virus displayed a significant reduction in FPR values at the end of treatment. Fast changes in the composition of viral populations were observed in all patients after 10 days of maraviroc (MVC) monotherapy treatment, and a complete replacement of viral quasiespecies was found in 3/10 patients carrying R5-using viruses and 4/10 patients carrying DM/X4-using viruses.IMPORTANCE Initiation of treatment with maraviroc requires previous determination of viral tropism by genotypic or phenotypic methods because of the risk of treatment failure and selection of DM/X4-tropic variants. In this study, we confirm previous work showing that the virologic response to maraviroc is independent of basal tropism. By deep-sequencing analysis, we determined that fast changes in viral populations were due to the emergence of minority variants in some patients whereas in others generation of new strains was detected. The risk of DM/X4 selection was very low as FPR values remained stable, and only one patient showed a detrimental switch to DM/X4 variants. Our data show that some DM/X4 viruses are sensitive to maraviroc treatment probably because only a low proportion of DM/X4 viruses use preferentially the X4 receptor and contain authentically maraviroc-resistant viruses that are not accurately detected by current assays.We thank Olga Palao (AIDS Immunopathogenesis Unit) and A. Zaballos (Genomics Unit, Instituto de Salud Carlos III) for their secretarial and technical assistance, respectively. We also greatly appreciate our patients for their willingness to participate.S

Harmful algae diversity from a coastal upwelling system detected by highthroughput sequencing

12 pages, 6 figures.-- This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY)[Introduction] In recent years, environmental DNA (eDNA) amplicon sequencing has been used to unveil plankton diversity in the field. Nevertheless, molecular methods, such as this, are rarely used in harmful algal bloom (HAB) monitoring programs, which mainly rely on morphological identification by conventional light microscopy. [Methods] The present study focused on a shallow marine environment (Ría de Vigo, Northwest Spain), where sediment and plankton samples were collected from 2016 to 2018. [Results] The application of eDNA amplicon sequencing allowed us to simultaneously detect 25 potential harmful species (mainly diatoms and dinoflagellates) included in the IOC-UNESCO Taxonomic Reference List of Harmful Microalgae. Among these, causative agents of amnesic shellfish poisoning (Pseudo-nitzschia spp.) paralytic shellfish poisoning (Gymnodinium catenatum and Alexandrium minutum), azaspiracid producers (Azadinium poporum) and ichthyotoxic haptophytes (Chrysochromulina leadbeateri), were identified. Some toxic microalgae were better represented in sediment (e.g., Pseudo-nitzschia pungens, Gymnodinium catenatum) or planktonic fractions (e.g., Pseudo-nitzschia, Gymnodinium smaydae), confirming the importance of including both sediment and plankton fractions in eDNA monitoring studies. Despite the limitations of sequencing short amplicons, it was possible to discern in this study six Pseudo-nitzschia species and associate each of them with each seasonal peak produced in summer periods. Furthermore, several species previously unreported in Ría de Vigo (Pseudo-nitzschia turgidula, Chrysochromulina leadbeateri, Azadinium poporum) could be detected. [Discusion] These results point out the application of eDNA amplicon sequencing to expand our knowledge about harmful species in HAB monitoring programs and early warning systems for low abundant and rare taxaThis work was supported by Fondo Europeo de Desarrollo Regional FEDER, Interreg España – Portugal (POCTEP) 2014-[20200474_BLUEBIOLAB], VIVALDI [678589] (EU H2020), Ministerio de Economía y Competitividad, Spain [CTM2017-83362-R] and Consellería de Economía, Emprego e Industria–GAIN, Xunta de Galicia [IN607B 2022/13]Peer reviewe

Bioactive compounds from marine phytoplankton (Project BLUEBIOLAB)

Poster.-- 19th International Conference on Harmful Algae, october 10-15 2021The objectives of the BLUEBIOLAB (Interreg-POCTEP; Spain-Portugal) are the creation of a transboundary laboratory of scientific excellence in marine biotechnology, to reinforce and internationalize the R & D capacities in the territory. For this purpose, several subprojects were launched for the period 2020 - 2022, related with marine biotechnology, aquaculture and biodiversity. The Oceanographic Center of Vigo (IEO, CSIC) participates in one of these initiatives, entitled ¿Bioactive compounds from marine photosynthetic organisms and biomedical potential¿, and led by UMINHO (Braga, Portugal) in consortium with CIIMAR, UdV and IIM-CSIC. Among the objectives of this initiative is the obtaining of microalgae cell extracts to search for bioactive compounds by means of biological activity assays. At present, cultures of 35 species of microalgae belonging to 7 classes (from the CCVIEO culture collection at IEO) have been performed: Dinophyceae class (22 species), Bacillariophyceae (4), Cryptophyceae (3), Prasinophyceae (1), Raphidophyceae (1), Euglenophyceae (1), Prymnesiophyceae (1), Dictyochophyceae (1), and Chlorophyceae (1). The cultivation parameters were selected to optimize the growth of each species. Culture biomass harvested for each strain was extracted following a protocol based on H2O - MeOH and CH2Cl2-MeOH. Final fractions obtained were solubilized in DMSO and kept at -80 ºC until carrying out the corresponding bioassays. In this regard, the antiviral activity is now being evaluated against spring viraemia of carp virus (SVCV), and antibacterial and anti-obesity activities will be subsequently evaluated. The bioassays will be performed using two approaches: in vitro cell cultures and in vivo assays using zebrafish as animal model. Those extracts showing biological activity will be analyzed by liquid chromatography coupled to mass spectrometry for compound identification.This research was funded by the BLUEBIOLAB project which is co-financed by the European Regional Development Fund FEDER through the Interreg V Spain-Portugal Program (POCTEP) 2014-2020N

Papel de la proteína de señalización agouti (ASP) en la determinación del patrón de pigmentación en peces. Aproximación molecular, celular y bioquímica mediante la utilización de sistemas de expresión diferencial

Una de las adaptaciones cromáticas de mayor éxito en vertebrados es la presencia de un patrón dorso/ventral de coloración en el cual la piel dorsal es Oscura mientras que la piel ventral es clara. En los peces, al igual que en otros vertebrados, la determinación del patrón de pigmentación se inicia a partir de la diferenciación de células pluripotentes de la cresta neural en estadios tempranos del desarrollo larvario. La señalización de las melanocortinas via receptor de tipo 1 (MClR) es clave en la determinación genética de la pigmentación en peces. La activación de receptor por la hormona estimulante de los melanocitos (a-MSH) promueve la diferenciación de las células precursoras hacia melanocitos, su profileración y la slntesis de melanina. Atlpicarnente, el MClR es también regulado por un antagonista endógeno, la proteína de sellalización agouti (ASP). En peces, ASP se expresa diferencialmente en la piel de la región ventral, que habitualmente muestra tonalidades claras en contraste con la región dorsal. Varias hipótesis defienden que este antagonismo sobre MCIR es responsable del patrón de pigmentación dorso-ventral en peces. Oicha polaridad en el patrón de pigmentación es extrema en los peces planos que habitualmente presentan problemas de hiper- o hipopigmentación en cultivo. En este estudio, presentamos la identificación y caracterización molecular de la proteína de sel\alización agouti (ASP), en el pez cebra y en el rodaballo (Scophthalmus maximus L.), su importancia en la determinación del patrón de pigmentación y su posible implicación en la inducción de malformaciones pigmentarias mediante el uso de técnicas de ADN recombinante y de transferencia génica.Este trabajo ha sido financiado gracias a la concesión de los contratos JAEDoc (IIM-CSIC) y Ramón y Cajal (MEC-CSIC) a RMC y JR respectivamente, y al proyecto MICIN AGL2008-00392/ACU.Peer Reviewe

What drives the number of high-risk human papillomavirus types in the anal canal in HIV-positive men who have sex with men?

We estimated the effect of sexual behavior, age, and immunodeficiency on the number of high-risk human papillomavirus (HR-HPV) types in the anal canal among human immunodeficiency virus-positive men who have sex with men (MSM). Anal samples were genotyped with the Linear Array HPV Genotyping Test, and risk factors were investigated with Poisson regression. Of 586 MSM, 69% were Spanish, and 25.6% were Latin American; the median age was 34.9 years (interquartile range [IQR], 30.1-40.8). The median number of recent sex partners was 6 (IQR, 2-24 sex partners), and the median CD4(+) T-cell count was 531.5 cells/mm(3) (IQR, 403-701 cells/mm(3)). The prevalence of any and multiple HR-HPV infections was 83.4% and 60.5%, respectively. The most common types were HPV-16 (42%), HPV-51 (24%), HPV-39 (23.7%), and HPV-59 (23.5%). Age had a statistically significant, nonlinear association with the number of types, with the highest number detected around 35 years of age (P <.001). The number of recent sex partners had a statistically significant, fairly linear association on the log scale (P = .033). The high prevalence of HR-HPV types is associated with recent sexual behavior and ag

Prolonged administration of maraviroc reactivates latent HIV in vivo but it does not prevent antiretroviral-free viral rebound.

Human immunodeficiency virus (HIV) remains incurable due to latent viral reservoirs established in non-activated CD4 T cells that cannot be eliminated via antiretroviral therapy. Current efforts to cure HIV are focused on identifying drugs that will induce viral gene expression in latently infected cells, commonly known as latency reversing agents (LRAs). Some drugs have been shown to reactivate latent HIV but do not cause a reduction in reservoir size. Therefore, finding new LRAs or new combinations or increasing the round of stimulations is needed to cure HIV. However, the effects of these drugs on viral rebound after prolonged treatment have not been evaluated. In a previous clinical trial, antiretroviral therapy intensification with maraviroc for 48 weeks caused an increase in residual viremia and episomal two LTR-DNA circles suggesting that maraviroc could reactivate latent HIV. We amended the initial clinical trial to explore additional virologic parameters in stored samples and to evaluate the time to viral rebound during analytical treatment interruption in three patients. Maraviroc induced an increase in cell-associated HIV RNA during the administration of the drug. However, there was a rapid rebound of viremia after antiretroviral therapy discontinuation. HIV-specific T cell response was slightly enhanced. These results show that maraviroc can reactivate latent HIV in vivo but further studies are required to efficiently reduce the reservoir size