Additional file 1 of Immunoescape of HIV-1 in Env-EL9 CD8 + T cell response restricted by HLA-B*14:02 in a Non progressor who lost twenty-seven years of HIV-1 control

Additional file 1. Oligonucleotides used for amplification. Table containing all the oligonucleotides used for the different PCR assays described in the Methods.Spanish National Research Council (CSIC) Instituto de Salud Carlos III (ISCIII) Spanish Government Xunta de Galicia Ministerio de Economía, Industria y Competitividad, Gobierno de España RIS-RETIC ISCIII RETIC Catalan Government and the European Social Fund.Peer reviewe

Isolation of Novel anti-SIV Monoclonal Antibodies from Rhesus Macaques

Almost forty years after the beginning of the AIDS epidemic, we do not have a vaccine or cure for HIV. While the discovery of antiretroviral therapy (ART) has ameliorated suffering from this disease, new strategies are still needed. Recently, the scientific community has turned its attention to broadly neutralizing monoclonal antibodies (bnmAbs) to prevent HIV acquisition and control viral replication. Human HIV-specific bnmAbs have been tested in preclinical trials using the chimeric simian/human immunodeficiency virus (SHIV) model in Indian rhesus macaques (RMs). Unfortunately, this artificial virus has several limitations and may not closely model HIV infection as well as the pathogenic simian immunodeficiency virus mac239 (SIVmac239) does. Thus, studies using SIV and SIV-specific mAbs might be more useful in understanding the role of Ab therapy in HIV-infected humans.&nbsp;The paucity of bona fide rhesus SIVmac239-specific mAbs has restricted our ability to study Abs for prophylaxis or therapy in the SIV-infected Indian RM model. Here, we describe the isolation of novel virus-specific mAbs from rhesus plasmablasts responding to viral replication in vivo, and from bone marrow-derived plasma cells. In the first instance, we induced viral replication by either CD8β+ T cell depletion or ART interruption in SIVmac239-infected Indian RMs with high neutralization Ab titers against this pathogenic clone. Second, we isolated SIVmac239 SOSIP.664-specific plasma cells from the bone marrow using a fluorescent microscope and a micromanipulator. We then determined the binding specificity of the 24 SIV-specific mAbs. Although we did not isolate any mAbs that neutralized SIVmac239, we identified several mAbs that neutralized 100% of the tier 1 SIVmac316 clone. Interestingly, most mAbs bound to primary rhesus CD4+ T cells infected with SIVmac239, and some induced Ab-dependent cellular cytotoxicity (ADCC), indicating that it is possible that these mAbs might confer antiviral immunity in vivo.&nbsp;</p

Use of Vaginal Dinoprostone (PGE₂) in Patients with Premature Rupture of Membranes (PROM) Undergoing Induction of Labor: A Comparative Study

Purpose: To evaluate the effect and safety of vaginal dinoprostone in pregnant women with PROM who undergo induction of labor (IoL). Materials and Methods: Prospective observational study conducted at La Mancha Centro hospital from 1 February 2019, to 30 August 2020. Obstetric and neonatal variables of 94 pregnant women with PROM who underwent IoL with vaginal dinoprostone were analyzed, and the results were compared with 330 patients without PROM who also underwent IoL. Bivariate and multivariate analyses were performed using binary and multiple linear regression. Results: A total of 424 women were included in this study. A greater response to cervical ripening (Bishop score > 6) with PGE2 was observed in the PROM group (odds ratio (OR) 2.73, 95% confidence interval (CI) 1.50–4.99, p = 0.001), as well as a shorter total duration of IoL (mean difference (MD) 2823.37 min (min), 95% CI 1257.30–4389.43, p n = 27) of patients in the PROM group vs. 34.2% (n = 113) of patients in the non-PROM group, with no significant differences (OR 0.87%, 95% CI 0.47–1.60, p = 0.652). There were no significant differences in changes in the cardiotocographic record (CTG), postpartum hemorrhage (PPH), uterine rupture, or adverse neonatal outcomes between the two groups. Conclusions: The use of vaginal dinoprostone in pregnant women undergoing IoL with PROM is safe for the mother and the fetus, shortens the total delivery time, and does not increase the risk of cesarean section compared with pregnant women undergoing IoL without PROM

Induction of Labor with Vaginal Dinoprostone (PGE2) in Patients with a Previous Cesarean Section: Obstetric and Neonatal Outcomes

Background: Vaginal dinoprostone (PGE2) is currently used as the prostaglandin of choice in many obstetric units. However, few studies have evaluated its safety, especially in women who previously had a cesarean section. Objective: To evaluate the efficacy and safety of PGE2 in pregnant women who are undergoing induction of labor (IOL), and who have had a previous cesarean section. Materials and Methods: A prospective observational study was conducted in La Mancha Centro Hospital in Alcázar de San Juan, Spain, from 1 February 2019 to 30 August 2020. Obstetric and neonatal outcomes, following IOL with PGE2, in 47 pregnant women who wanted a trial of labor after cesarean (TOLAC), and 377 pregnant women without a history of cesarean section, were analyzed. The outcomes were analyzed by bivariate and multivariate analyses using binary and multiple linear regression. Results: A total of 424 women were included in this study. The percentage of cesarean sections in the TOLAC group was 44.7% (21), compared with 31.6% (119) in the group without a history of cesarean section (adjusted odds ratio: 1.4; 95% CI: 0.68–2.86). In the multivariate analysis, no statistically significant differences were observed between both groups for obstetric and neonatal outcomes (p &gt; 0.05). However, two uterine ruptures (4.3%) occurred in the group of patients with a history of cesarean section who underwent IOL with PGE2. Conclusions: The induction of labor with vaginal dinoprostone (PGE2), in patients with a previous history of cesarean section, was not associated with worse obstetric or neonatal outcomes compared with the group of patients without a history of cesarean section in our study sample. However, further research is needed regarding this IOL method, and it should be used with caution in this population group

Use of Vaginal Dinoprostone (PGE2) in Patients with Premature Rupture of Membranes (PROM) Undergoing Induction of Labor: A Comparative Study

Purpose: To evaluate the effect and safety of vaginal dinoprostone in pregnant women with PROM who undergo induction of labor (IoL). Materials and Methods: Prospective observational study conducted at La Mancha Centro hospital from 1 February 2019, to 30 August 2020. Obstetric and neonatal variables of 94 pregnant women with PROM who underwent IoL with vaginal dinoprostone were analyzed, and the results were compared with 330 patients without PROM who also underwent IoL. Bivariate and multivariate analyses were performed using binary and multiple linear regression. Results: A total of 424 women were included in this study. A greater response to cervical ripening (Bishop score &gt; 6) with PGE2 was observed in the PROM group (odds ratio (OR) 2.73, 95% confidence interval (CI) 1.50&ndash;4.99, p = 0.001), as well as a shorter total duration of IoL (mean difference (MD) 2823.37 min (min), 95% CI 1257.30&ndash;4389.43, p &lt; 0.001). Cesarean sections were performed in 28.7% (n = 27) of patients in the PROM group vs. 34.2% (n = 113) of patients in the non-PROM group, with no significant differences (OR 0.87%, 95% CI 0.47&ndash;1.60, p = 0.652). There were no significant differences in changes in the cardiotocographic record (CTG), postpartum hemorrhage (PPH), uterine rupture, or adverse neonatal outcomes between the two groups. Conclusions: The use of vaginal dinoprostone in pregnant women undergoing IoL with PROM is safe for the mother and the fetus, shortens the total delivery time, and does not increase the risk of cesarean section compared with pregnant women undergoing IoL without PROM

Differential epigenetic regulation between the alternative promoters, PRDM1 alpha and PRDM1 beta, of the tumour suppressor gene PRDM1 in human multiple myeloma cells

Multiple myeloma (MM) is a B-cell neoplasm that is characterized by the accumulation of malignant plasma cells in the bone marrow. The transcription factor PRDM1 is a master regulator of plasma cell development and is considered to be an oncosuppressor in several lymphoid neoplasms. The PRDM1 beta isoform is an alternative promoter of the PRDM1 gene that may interfere with the normal role of the PRDM1 alpha isoform. To explain the induction of the PRDM1 beta isoform in MM and to offer potential therapeutic strategies to modulate its expression, we characterized the cis regulatory elements and epigenetic status of its promoter. We observed unexpected patterns of hypermethylation and hypomethylation at the PRDM1 alpha and PRDM1 beta promoters, respectively, and prominent H3K4me1 and H3K9me2 enrichment at the PRDM1 beta promoter in non-expressing cell lines compared to PRDM1 beta-expressing cell lines. After treatment with drugs that inhibit DNA methylation, we were able to modify the activity of the PRDM1 beta promoter but not that of the PRDM1 alpha promoter. Epigenetic drugs may offer the ability to control the expression of the PRDM1 alpha/PRDM1 beta promoters as components of novel therapeutic approaches

Novel Spike-stabilized trimers with improved production protect K18-hACE2 mice and golden Syrian hamsters from the highly pathogenic SARS-CoV-2 Beta variant

Most COVID-19 vaccines are based on the SARS-CoV-2 Spike glycoprotein (S) or their subunits. However, S shows some structural instability that limits its immunogenicity and production, hampering the development of recombinant S-based vaccines. The introduction of the K986P and V987P (S-2P) mutations increases the production and immunogenicity of the recombinant S trimer, suggesting that these two parameters are related. Nevertheless, S-2P still shows some molecular instability and it is produced with low yield. Here we described a novel set of mutations identified by molecular modeling and located in the S2 region of the S-2P that increase its production up to five-fold. Besides their immunogenicity, the efficacy of two representative S-2P-based mutants, S-29 and S-21, protecting from a heterologous SARS-CoV-2 Beta variant challenge was assayed in K18-hACE2 mice (an animal model of severe SARS-CoV-2 disease) and golden Syrian hamsters (GSH) (a moderate disease model). S-21 induced higher level of WH1 and Delta variants neutralizing antibodies than S-2P in K18-hACE2 mice three days after challenge. Viral load in nasal turbinate and oropharyngeal samples were reduced in S-21 and S-29 vaccinated mice. Despite that, only the S-29 protein protected 100% of K18-hACE2 mice from severe disease. When GSH were analyzed, all immunized animals were protected from disease development irrespectively of the immunogen they received. Therefore, the higher yield of S-29, as well as its improved immunogenicity and efficacy protecting from the highly pathogenic SARS-CoV-2 Beta variant, pinpoint the S-29 mutant as an alternative to the S-2P protein for future SARS-CoV-2 vaccine development

Immunisation efficacy of a stabilised SARS-CoV-2 spike glycoprotein in two geriatric animal models

Altres ajuts: Grífols (CBIG), Direcció General de Recerca i Innovació en Salut (SLD0015 and SDL0016) and Crowdfunding "yomecorono.com".Age is associated with reduced efficacy of vaccines and linked to higher risk of severe COVID-19. Here we determined the impact of ageing on the efficacy of a SARS-CoV-2 vaccine based on a stabilised Spike glycoprotein (S-29) that had previously shown high efficacy in young animals. Thirteen to 18-month-old golden Syrian hamsters (GSH) and 22-23-month-old K18-hCAE2 mice were immunised twice with S-29 protein in AddaVax TM adjuvant. GSH were intranasally inoculated with SARS-CoV-2 either two weeks or four months after the booster dose, while all K18-hACE2 mice were intranasally inoculated two weeks after the second immunisation. Body weight and clinical signs were recorded daily post-inoculation. Lesions and viral load were investigated in different target tissues. Immunisation induced seroconversion and production of neutralising antibodies; however, animals were only partially protected from weight loss. We observed a significant reduction in the amount of viral RNA and a faster viral protein clearance in the tissues of immunized animals. Infectious particles showed a faster decay in vaccinated animals while tissue lesion development was not altered. In GSH, the shortest interval between immunisation and inoculation reduced RNA levels in the lungs, while the longest interval was equally effective in reducing RNA in nasal turbinates; viral nucleoprotein amount decreased in both tissues. In mice, immunisation was able to improve the survival of infected animals. Despite the high protection shown in young animals, S-29 efficacy was reduced in the geriatric population. Our research highlights the importance of testing vaccine efficacy in older animals as part of preclinical vaccine evaluation

Immunization with V987H-stabilized Spike glycoprotein protects K18-hACE2 mice and golden Syrian hamsters upon SARS-CoV-2 infection

Safe and effective severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines are crucial to fight against the coronavirus disease 2019 pandemic. Most vaccines are based on a mutated version of the Spike glycoprotein [K986P/V987P (S-2P)] with improved stability, yield and immunogenicity. However, S-2P is still produced at low levels. Here, we describe the V987H mutation that increases by two-fold the production of the recombinant Spike and the exposure of the receptor binding domain (RBD). S-V987H immunogenicity is similar to S-2P in mice and golden Syrian hamsters (GSH), and superior to a monomeric RBD. S-V987H immunization confer full protection against severe disease in K18-hACE2 mice and GSH upon SARS-CoV-2 challenge (D614G or B.1.351 variants). Furthermore, S-V987H immunized K18-hACE2 mice show a faster tissue viral clearance than RBD- or S-2P-vaccinated animals challenged with D614G, B.1.351 or Omicron BQ1.1 variants. Thus, S-V987H protein might be considered for future SARS-CoV-2 vaccines development. In this study, the authors report a mutation that increases the production of recombinant SARS-CoV-2 Spike and exposure of the RBD. In animal models, a Spike-based vaccine containing the mutation induces strong immunogenicity, provides protection from disease and results in faster tissue viral clearance