Latency reversal agents affect differently the latent reservoir present in distinct CD4+ t subpopulations

Latency reversal agents (LRAs) have proven to induce HIV-1 transcription in vivo but are ineffective at decreasing the size of the latent reservoir in antiretroviral treated patients. The capacity of the LRAs to perturb the viral reservoir present in distinct subpopulations of cells is currently unknown. Here, using a new RNA FISH/flow ex vivo viral reactivation assay, we performed a comprehensive assessment of the viral reactivation capacity of different families of LRAs, and their combinations, in different CD4 T cell subsets. We observed that a median of 16.28% of the whole HIV-reservoir induced HIV-1 transcripts after viral reactivation, but only 10.10% of these HIV-1 RNA cells produced the viral protein p24. Moreover, none of the LRAs were powerful enough to reactivate HIV-1 transcription in all CD4 T cell subpopulations. For instance, the combination of Romidepsin and Ingenol was identified as the best combination of drugs at increasing the proportion of HIV-1 RNA cells, in most, but not all, CD4 T cell subsets. Importantly, memory stem cells were identified as highly resistant to HIV-1 reactivation, and only the combination of Panobinostat and Bryostatin-1 significantly increased the number of cells transcribing HIV within this subset. Overall, our results validate the use of the RNA FISH/flow technique to assess the potency of LRAs among different CD4 T cell subsets, manifest the intrinsic differences between cells that encompass the latent HIV reservoir, and highlight the difficulty to significantly impact the latent infection with the currently available drugs. Thus, our results have important implications for the rational design of therapies aimed at reversing HIV latency from diverse cellular reservoirs

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

ANTI M. leprae IgM ANTIBODY DETERMINATION BY ULTRAMICROIMMUNOENZYMATIC (UMELISA HANSEN) FOR THE DIAGNOSIS AND MONITORING LEPROSY

The relationship between the IgM antibody response, antigenic load as well as the clinical improvement after chemotherapy was studied in order to obtain useful data for the early diagnosis and monitoring leprosy. A level of 82% (94/115) agreement was obtained between IgM UMELISA HANSEN and slitskin smear examination. Discrepant results were observed in 16 patients who showed positive IgM response despite negative by the skin smear examination. In these patients, the IgM response was seen to be associated to the early signal for bacilli recurrence in the skin. In one of these patients the presence of bacilli was demonstrated in the skin, two months after IgM antibodies being detected by UMELISA HANSEN. Also in one of the treated patients positive by both diagnostic techniques, a remarkable decrease in the IgM antibody levels was seen, correlating with a significant clinical improvement. Moreover it was found a direct relationship between the IgM antibody response and bacterial antigenic load, regardless the time elapsed in the disease's evolution

UltramicroELISA para la deteccion de anticuerpos IgM al Mycobacterium leprae utilizando muestras de sangre seca UltramicroELISA assay for the detection of IgM antibodies to Mycobacterium leprae using eluates of dried blood spots

En este trabajo se establecen las condiciones optimas para la detección de anticuerpos IgM al glicolipido fenólico-I (GF-I) en muestras de sangre en papel de filtro utilizando el UltramicroELISA HANSEN y la tecnologia SUMA. Se estudiaron 300 donantes de sangre y 58 pacientes leprosos. Para estas dos poblaciones se compararon los resultados de muestras de sangre seca colectadas en papel de filtro SS-2992 con los de suero, y se obtuvo una correlación de 0.919 para donantes de sangre, 0.969 para pacientes y 0.954 para el total de las dos poblaciones. Se obtuvo una coincidencia de 98% en pacientes y 96% en donantes. En la población de pacientes la sensibilidad fue de 93% y la especificidad de 100% para las muestras de sangre seca evaluadas por el UMELISA HANSEN, con respecto a las muestras de suero analizadas por esta misma tecnica.<br>In this work the adequate conditions for the detection of IgM antibodies to PGL-I in eluates from dried blood spots using the UMELISA HANSEN and the SUMA technology are established. A total of 300 blood donors and 58 leprosy patients samples were studied. For both populations, in the previously established conditions, we compared the results of the eluates from dried blood spots with the serum samples, and was obtained a correlation of 0.919 in blood donors, 0.969 in patients, and 0.954 for the total of both populations. It was also obtained a level of agreemnet of 98% in patients and 96% in blood donors. In the patients population was found a sensitivity of 93% and an specificity of 100% for the eluates of dried blood spots evaluated by the UMELISA HANSEN in the comparison with the serum samples analized by the same assay

UltramicroELISA para la deteccion de anticuerpos IgM al Mycobacterium leprae utilizando muestras de sangre seca

En este trabajo se establecen las condiciones optimas para la detección de anticuerpos IgM al glicolipido fenólico-I (GF-I) en muestras de sangre en papel de filtro utilizando el UltramicroELISA HANSEN y la tecnologia SUMA. Se estudiaron 300 donantes de sangre y 58 pacientes leprosos. Para estas dos poblaciones se compararon los resultados de muestras de sangre seca colectadas en papel de filtro SS-2992 con los de suero, y se obtuvo una correlación de 0.919 para donantes de sangre, 0.969 para pacientes y 0.954 para el total de las dos poblaciones. Se obtuvo una coincidencia de 98% en pacientes y 96% en donantes. En la población de pacientes la sensibilidad fue de 93% y la especificidad de 100% para las muestras de sangre seca evaluadas por el UMELISA HANSEN, con respecto a las muestras de suero analizadas por esta misma tecnica.In this work the adequate conditions for the detection of IgM antibodies to PGL-I in eluates from dried blood spots using the UMELISA HANSEN and the SUMA technology are established. A total of 300 blood donors and 58 leprosy patients samples were studied. For both populations, in the previously established conditions, we compared the results of the eluates from dried blood spots with the serum samples, and was obtained a correlation of 0.919 in blood donors, 0.969 in patients, and 0.954 for the total of both populations. It was also obtained a level of agreemnet of 98% in patients and 96% in blood donors. In the patients population was found a sensitivity of 93% and an specificity of 100% for the eluates of dried blood spots evaluated by the UMELISA HANSEN in the comparison with the serum samples analized by the same assay

Expression of CD20 after viral reactivation renders HIV-reservoir cells susceptible to Rituximab

The identification of exclusive markers to target HIV-reservoir cells will represent a significant advance in the search for therapies to cure HIV. Here, we identify the B lymphocyte antigen CD20 as a marker for HIV-infected cells in vitro and in vivo. The CD20 molecule is dimly expressed in a subpopulation of CD4-positive (CD4) T lymphocytes from blood, with high levels of cell activation and heterogeneous memory phenotypes. In lymph node samples from infected patients, CD20 is present in productively HIV-infected cells, and ex vivo viral infection selectively upregulates the expression of CD20 during early infection. In samples from patients on antiretroviral therapy (ART) this subpopulation is significantly enriched in HIV transcripts, and the anti-CD20 monoclonal antibody Rituximab induces cell killing, which reduces the pool of HIV-expressing cells when combined with latency reversal agents. We provide a tool for targeting this active HIV-reservoir after viral reactivation in patients while on ART

COVID-19 Follow-App. Mobile App-Based Monitoring of COVID-19 Patients after Hospital Discharge : A Single-Center, Open-Label, Randomized Clinical Trial

Introduction: In the midst of a pandemic, apps can be used to provide close follow-up, ensure that patients are monitored at home, avoid excessive pressure on medical facilities, prevent the movement of people (both patients and health professionals), and reduce the risk of infection. Objective: To adapt and validate the use of a smartphone application for outpatient follow-up of COVID-19 patients after hospital discharge. Methods: We conducted an open-label clinical trial at Hospital Universitari Vall d'Hebron in Barcelona, Spain. Patients were randomly assigned in a 1:1 ratio to be followed by the Farmalarm app or by their primary care center. The primary endpoint was the reduction in the need for in-person return visits. Results: From 31 March to 4 May 2020, 150 patients were enrolled in the study at hospital discharge: 74 patients were randomized to the experimental group, and 76 to the control group. All patients in the control group and all except for six in the experimental group completed the study. During hospitalization, before study inclusion, all but 4 (97.3%) had viral pneumonia, 91 (60.7%) required supplemental oxygen, and 16 (10.7%) required intensive care unit (ICU) admission. COVID-19-related return visits to the emergency department were significantly higher in the control group (7.9% vs. 0%; p = 0.028) in the per-protocol analysis. Telephone consultations with the emergency department were performed by 12 (15.8%) patients in the control group and 0 (0%) in the experimental group (p < 0.001). Satisfaction with outpatient monitoring was rated higher by the experimental group (5 vs. 4 points; p < 0.001). Conclusions: Following COVID-19 hospital discharge, home follow-up via a mobile app was effective in reducing in-person return visits without undermining patient satisfaction or perception of health, compared with standard follow-u