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Nanoparticles derived from plant viruses play an important role in nanomedicine due to their biocompatibility, self-assembly and easily-modifiable surface. In this study, we developed a novel platform for increasing antibody sensing using viral nanoparticles derived from turnip mosaic virus (TuMV) functionalized with SARS-CoV-2 receptor binding domain (RBD) through three different methods: chemical conjugation, gene fusion and the SpyTag/SpyCatcher technology. Even though gene fusion turned out to be unsuccessful, the other two constructs were proven to significantly increase antibody sensing when tested with saliva of patients with different infection and vaccination status to SARS-CoV-2. Our findings show the high potential of TuMV nanoparticles in the fields of diagnostics and immunodetection, being especially attractive for the development of novel antibody sensing devices., Ministerio de Universidades (España), Comunidad de Madrid, Universidad Complutense de Madrid, Depto. de Genética, Fisiología y Microbiología, Fac. de Ciencias Biológicas, TRUE, pub

2024 Acuerdos transformativos CRUE-CSIC, Both EphB2- and EphB3-deficient mice exhibit profound histological alterations in the thymic epithelial network but few changes in T-cell differentiation, suggesting that this organization would be sufficient to produce functional T lymphocytes. Also, other antigen-presenting cells involved in immunological education could substitute the thymic epithelium. Accordingly, we found an increased frequency of plasmacytoid dendritic cells but not of conventional dendritic cells, medullary fibroblasts or intrathymic B lymphocytes. In addition, there are no lymphoid infiltrates in the organs of mutant mice nor do they contain circulating autoantibodies. Furthermore, attempts to induce arthritic lesions after chicken type II collagen administration fail totally in EphB2-deficient mice whereas all WT and half of the immunized EphB3−/− mice develop a typical collagen-induced arthritis. Our results point out that Th17 cells, IL4-producing Th2 cells and regulatory T cells are key for the induction of disease, but mutant mice appear to have deficits in T cell activation or cell migration properties. EphB2−/− T cells show reduced in vitro proliferative responses to anti-CD3/anti-CD28 antibodies, produce low levels of anti-type II collagen antibodies, and exhibit low proportions of T follicular helper cells. On the contrary, EphB3−/− lymph node cells respond accurately to the different immune stimuli although in lower levels than WT cells but show a significantly reduced migration in in vitro transwell assays, suggesting that no sufficient type II collagen-dependent activated lymphoid cells reached the joints, resulting in reduced arthritic lesions., Unión Europea. NextGenerationEU. Plan de Recuperación Transformación y Resiliencia, Ministerio de Ciencia, Innovación y Universidades, Instituto de Salud Carlos III, Depto. de Biología Celular, Fac. de Ciencias Biológicas, TRUE, pub

Introduction: Recurrent urinary tract infections (RUTIs) and recurrent vulvovaginal candidiasis (RVVCs) represent major healthcare problems all over the world. Antibiotics and antifungals are widely used for such infectious diseases, which is linked with microbial resistances and microbiota deleterious effects. The development of novel approaches for genitourinary tract infections (GUTIs) such as trained immunity-based vaccines (TIbV) is therefore highly required. MV140 is a sublingual whole-cell heat-inactivated polybacterial preparation with demonstrated clinical efficacy for RUTIs. The sublingual heat-inactivated Candida albicans vaccine V132 has been developed for RVVCs. We previously showed that the combination of MV140 and V132 promotes potent Th1/Th17 and regulatory T-cell responses against antigens contained in the formulation and unrelated antigens. The specific contribution of each preparation to such effects and the underlying molecular mechanisms remain incompletely understood. Methods: PBMC and monocytes were isolated from healthy donors and in vitro stimulated with V132, MV140 or MV140/V132. After 6 days of resting, cells were reestimulated with LPS and MV140. Analysis of cytokine production by ELISA, Seahorse assays for functional metabolic experiments and chromatin immunoprecipitation assays were performed. BALB/c mice were intraperitoneally and sublingually immunized with V132. Results: We uncover that V132 induces trained immunity in human PBMCs and purified monocytes, significantly increasing the responses triggered by purified monocytes, significantly increasing the responses triggered by subsequent stimulation with MV140. Mechanistically, V132 drives metabolic rewiring towards increased glycolysis and oxidative phosphorylation and induces epigenetic reprogramming that enhances the transcription of the pro-inflammatory genes IL6 and TNFA. Splenocytes and peritoneal cells from V132-immunize mice show increased responses upon in vitro stimulation w, MINECO, The Netherlands Organization for Scientific Research, Netherlands Organization for Scientific Research, ERC Advanced Grant, Depto. de Bioquímica y Biología Molecular, Fac. de Ciencias Químicas, TRUE, pub

Funding This work was funded by EU H2020 program through ParaFishControl Project (634429) and by the European Research Council (ERC Consolidator Grant 2016 725061 TEMUBLYM). This publication reflects only the authors’ view and the European Union cannot be held responsible for any use that may be made of the information contained herein. MP was funded by a Ramón y Cajal Postdoctoral Research Fellowship (RYC2018-024049-I/AEI/10.13039/501100011033 %CITAVIPICKER£10.13039/501100011033££% & ESF), IE under APOSTD/2016/037 grant by the “Generalitat Valenciana” and RP was contracted under the PTA-Program from the Spanish Ministry of Science, Innovation and Universities (PTA2018-015315-I). We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI)., Passive immunization constitutes an emerging field of interest in aquaculture, particularly with the restrictions for antibiotic use. Enteromyxum leei is a myxozoan intestinal parasite that invades the paracellular space of the intestinal epithelium, producing a slow-progressing disease, leading to anorexia, cachexia and mortalities. We have previously demonstrated that gilthead sea bream (GSB, Sparus aurata) that survive E. leei infection become resistant upon re-exposure, and this resistance is directly related to the presence of high levels of specific IgM in serum. Thus, the current work was aimed to determine if passive immunization could help to prevent enteromyxosis in GSB and to study in detail the nature of these protective antibodies. Serum from a pool of resistant (SUR) or naïve (NAI) animals was intracoelomically injected 24 h prior to the E. leei-effluent challenge and at 9 days post-challenge (dpc). Effluent challenge lasted for 23 days, and then the injected groups were allocated in separate tanks with clean water. A non-lethal parasite diagnosis was performed at 56 dpc. At the final sampling (100 dpc), blood, serum and tissues were collected for histology, molecular diagnosis and the detection of circulating antibodies. In parallel, we performed an immunoglobulin repertoire analysis of the fish generating SUR and NAI sera. The results showed that, fish injected with parasite-specific antibodies (spAbs) became infected with the parasite, but showed lower disease signs and intensity of infection than the other groups, indicating a later establishment of the parasite. Repertoire analysis revealed that E. leei induced a polyclonal expansion of diverse IgM and IgT subsets that could be in part an evasion strategy of the parasite. Nonetheless, GSB was able to produce sufficient levels of parasite-spAbs to avoid re-infection of surviving animals and confer certain degree of protection upon passive transfer of antibodies. These results highlight the crucial, Depto. de Genética, Fisiología y Microbiología, Fac. de Ciencias Biológicas, TRUE, pub

Introduction: Given the wide heterogeneity of common variable immunodeficiency (CVID), several groups have proposed clinical and immunological classifications to better define follow-up and prognostic algorithms. The present study aims to validate recent clinical and laboratory algorithms, based on different combinations of CVID biomarkers, to provide more personalized treatment and follow-up strategies. Methods: We analysed clinical and immunological features of 80 patients with suspected or diagnosed CVID, in two reference centres of Portugal and Spain. Clinical manifestations were categorized into clinical phenotyping proposed by Chapel et al. [1] that included cytopenia; polyclonal lymphocytic infiltration; unexplained enteropathy; and no disease-related complications. Results: 76% of patients in our cohort entered one of the four categories of clinical phenotyping, without overlap (cytopenia; polyclonal lymphocytic infiltration; unexplained enteropathy; and no disease-related complications). The most prominent phenotype was "cytopenia" (40%) followed by "polyclonal lymphocytic infiltration" (19%). The remaining 24% patients of our cohort had overlap of 2 clinical phenotypes (cytopenia and unexplained enteropathy mainly). A delay of CVID diagnosis in more than 6 years presented 3.7-fold higher risk of developing lymphoproliferation and/or malignancy (p < 0.05), and was associated with increased CD8+CD45RO + T-lymphocytes (p < 0.05). An association between decreased switched-memory B cells with lymphoproliferation and malignancy was observed (p < 0.03 and p < 0.05, respectively). CD4 + T-lymphocytopenia correlated with autoimmune phenotype, with 30% prevalence (p < 0.05). HLA-DR7 expression was related to CVID onset in early life in our patients (13 vs 25 years), and DQ2.5 or DQ2.2 with unexplained enteropathy (p < 0.05). Conclusions: The phenotypic and genetic study is crucial for an adequate clinical orientation of CVID patients. In these two independent cohort, European Federation of Immunological Societies (EFIS), Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

2018 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. En esta publicación han participado: 1Department of Internal Medicine III, University Hospital, RWTH, Aachen, Germany; 2Department of Immunology, Ophtalmology & ORL,Complutense University School of Medicine, Madrid, Spain; 3 12 de Octubre Health Research Institute (imas12), Madrid, Spain; 4Department of Hepatobiliary Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing Jiangsu, China; 5Department of Anesthesiology and Pain Management, Shanghai East Hospital, Tongji University, Shanghai, China; 6Institute of Pathology, Braunschweig Hospital, Braunschweig, Germany, Background & Aims: Caspase 8 (CASP8) is the apical initiator caspase in death receptor-mediated apoptosis. Strong evidence for a link between death receptor signaling pathways and cholestasis has recently emerged. Herein, we investigated the role of CASP8-dependent and independent pathways during experimental cholestasis. Methods: Liver injury was characterized in a cohort of human sera (n = 28) and biopsies from patients with stage IV primary biliary cholangitis. In parallel, mice with either specific deletion of Casp8 in liver parenchymal cells (Casp8Dhepa) or hepatocytes (Casp8Dhep), and mice with constitutive Ripk3 (Ripk3 / ) deletion, were subjected to surgical ligation of the common bile duct (BDL) from 2 to 28 days. Floxed (Casp8fl/fl) and Ripk3+/+ mice were used as controls. Moreover, the pan-caspase inhibitor IDN-7314 was used, and cell death mechanisms were studied in primary isolated hepatocytes. Results: Overexpression of activated caspase 3, CASP8 and RIPK3 was characteristic of liver explants from patients with primary biliary cholangitis. Twenty-eight days after BDL, Casp8Dhepa mice showed decreased necrotic foci, serum aminotransferase levels and apoptosis along with diminished compensatory proliferation and ductular reaction. These results correlated with a decreased inflammatory profile and ameliorated liver fibrogenesis. A similar phenotype was observed in Ripk3-/- mice. IDN-7314 treatment decreased CASP8 levels but failed to prevent BDL-induced cholestasis, independently of CASP8 in hepatocytes. Conclusion: These findings show that intervention against CASP8 in liver parenchymal cells – specifically in cholangiocytes– might be a beneficial option for treating obstructive cholestasis, while broad pan-caspase inhibition might trigger undesirable side effects. Lay summary: Loss of caspase 8 – a protein involved in programmed cell death – in liver parenchymal cells protects against experimental cholestasis. Therefore, specific pharmacological interve, IZKF, Ministerio de Economía, Comercio y Empresa, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Recurrent respiratory tract infections (RRTIs) are the first leading cause of community- and nosocomial-acquired infections. Antibiotics remain the mainstay of treatment, enhancing the potential to develop antibiotic resistances. Therefore, the development of new alternative approaches to prevent and treat RRTIs is highly demanded. Daily sublingual administration of the whole heat-inactivated polybacterial preparation (PBP) MV130 significantly reduced the rate of respiratory infections in RRTIs patients, however, the immunological mechanisms of action remain unknown. Herein, we study the capacity of MV130 to immunomodulate the function of human dendritic cells (DCs) as a potential mechanism that contribute to the clinical benefits. We demonstrate that DCs from RRTIs patients and healthy controls display similar ex vivo immunological responses to MV130. By combining systems biology and functional immunological approaches we show that MV130 promotes the generation of Th1/Th17 responses via receptor-interacting serine/threonine-protein kinase-2 (RIPK2)- and myeloid-differentiation primary-response gene-88 (MyD88)-mediated signalling pathways under the control of IL-10. In vivo BALB/c mice sublingually immunized with MV130 display potent systemic Th1/Th17 and IL-10 responses against related and unrelated antigens. We elucidate immunological mechanisms underlying the potential way of action of MV130, which might help to design alternative treatments in other clinical conditions with high risk of recurrent infections., MINECO, INNPACTO, Depto. de Bioquímica y Biología Molecular, Fac. de Ciencias Químicas, TRUE, pub

MINECO, Ministerio de Ciencia e Innovación, CDTI, Depto. de Bioquímica y Biología Molecular, Fac. de Ciencias Químicas, TRUE, pub

*Department of Medicine III, University Hospital, Aachen, Germany; ‡Department of Radiology, Justus-Liebig University, Giessen, Germany; §Institute of Pathology, University Hospital Aachen, Germany; and Human Genetics Division, University of Southampton, Southampton General Hospital, Southampton, United Kingdom, BACKGROUND & AIMS: Disruption of the nuclear factor-kB (NF-kB) essential modulator (NEMO) in hepatocytesof mice (NEMOAhepa mice) results in spontaneous liver apoptosis and chronic liver disease involving inflammation, steatosis, fibrosis, and development of hepatocellular carcinoma. Activation of caspase-8 (Casp8) initiates death receptor-mediated apoptosis. We investigated the pathogenic role of this protease in NEMOAhepa mice or after induction of acute liver injury. METHODS: We created mice with conditional deletion of Casp8 in hepatocytes (Casp8Ahepa) and Casp8AhepaNEMOAhepa double knockout mice. Acute liver injury was induced by Fas-activating antibodies, lipopolysaccharides, or concanavalin A. Spontaneous hepatocarcinogenesis was monitored by magnetic resonance imaging. RESULTS: Hepatocyte-specific deletion of Casp8 protected mice from induction of apoptosis and liver injury by Fas or lipopolysaccharides but increased necrotic damage and reduced survival times of mice given concanavalin A. Casp8AhepaNEMOAhepa mice were protected against steatosis and hepatocarcinogenesis but had a separate, spontaneous phenotype that included massive liver necrosis, cholestasis, and biliary lesions. The common mechanism by which inactivation of Casp8 induces liver necrosis in both injury models involves the formation of protein complexes that included the adaptor protein Fas-associated protein with death domain and the kinases receptor-interacting protein (RIP) 1 and RIP3—these have been shown to be required for programmed necrosis. We demonstrated that hepatic RIP1 was proteolytically cleaved by Casp8, whereas Casp8 inhibition resulted in accumulation of RIP complexes and subsequent liver necrosis. CONCLUSIONS: Inhibition of Casp8 protects mice from hepatocarcinogenesis following chronic liver injury mediated by apoptosis of hepatocytes but can activate RIP-mediated necrosis in an inflammatory environment., Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, Deutsche Krebshilfe, Deutsche Forschungsgemeinschaf, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Over the last several years, next-generation sequencing and its recent push toward single-cell resolution have transformed the landscape of immunology research by revealing novel complexities about all components of the immune system. With the vast amounts of diverse data currently being generated, and with the methods of analyzing and combining diverse data improving as well, integrative systems approaches are becoming more powerful. Previous integrative approaches have combined multiple data types and revealed ways that the immune system, both as a whole and as individual parts, is affected by genetics, the microbiome, and other factors. In this review, we explore the data types that are available for studying immunology with an integrative systems approach, as well as the current strategies and challenges for conducting such analyses., Burroughs Wellcome Fund (D.B.), Depto. de Estadística y Ciencia de los Datos, Fac. de Estudios Estadísticos, TRUE, pub

Los factores reguladores del Interferón (IRF) pertenecen a una familia de factores de transcripción que incluye varios miembros que regulan la expresión de genes tanto proinflamatorios como antiinflamatorios. En este trabajo se estudió la respuesta de ratones con una mutación dirigida en IRF-2 (IRF-2(-/-)) a la inyección de lipopolisacárido (LPS) con el fin de evaluar la importancia de IRF-2 en la regulación de la endotoxicidad. Los ratones IRF-2(-/-) fueron altamente refractarios a la letalidad inducida por LPS. Aunque el ARNm del TNF-alfa hepático y el TNF-alfa circulante estaban significativamente elevados en los ratones IRF-2(-/-) tratados con LPS, los niveles de ARNm y proteínas de IL-1, IL-12 e IFN-gamma, así como de la proteína IL-6, fueron significativamente más bajos que los niveles observados en los ratones IRF-2(+/+) tratados con LPS. Los ratones IRF-2(-/-) también fueron más refractarios a TNF-alfa que los ratones control, resultado consistente con su sensibilidad disminuida al LPS. Sin embargo, no se observaron diferencias significativas en la expresión de ARNm de TNFR. Los niveles de ARNm de IL-12R beta 2 de ratones IRF-2(-/-) tratados con LPS fueron significativamente menores que los del grupo control después de 1, 6 y 8 h de tratamiento, lo que sugiere que tanto la disminución de IL-12 como la alteración de la expresión de IL-12R contribuyen a la escasez de IFN-gamma producido. Los ratones knockout de IRF-2 tampoco lograron mantener los niveles de ARNm inducibles por LPS de IRF-1 y de ICSBP, dos factores necesarios para la transcripción de IL-12, tal vez como resultado de la disminución de los niveles de IL-1 beta, IL-6 e IFN-gamma. También se analizaron secciones hepáticas de ratones IRF-2(+/+) e IRF-2(-/-) 6 h después de una inyección típicamente letal de LPS. Los ratones IRF-2(-/-) exhibieron un mayor número de células de Kupffer apoptóticas que los ratones de tipo salvaje, lo que sugiere un nuevo papel antiapoptótico para IRF-2. En conjunto, esto, National Institutes of Health, Depto. de Biología Celular, Fac. de Medicina, TRUE, pub

The cytoplasmic protein tumor progression locus 2 (TPL2), also known as cancer Osaka thyroid (Cot), or MAP3K8, is thought to have a significant role in a variety of cancers and illnesses and it is a key component in the activation pathway for the expression of inflammatory mediators. Despite the tight connection between inflammation and TPL2, its function has not been extensively studied in chronic liver disease (CLD), a major cause of morbidity and mortality worldwide. Here, we analyze more in detail the significance of TPL2 in CLD to shed light on the pathological and molecular transduction pattern of TPL2 during the progression of CLD. This might result in important advancements and enable progress in the diagnosis and treatment of CLD., Comunidad Autónoma de Madrid, Ministerio de Ciencia e Innovación, Comunidad Europea, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Acknowledgements We are grateful to our laboratory members and the Kennedy Institute of Rheumatology community for constructive scientific discussions, especially to James Felce, David Depoil, Jonathan Webber, Štefan Balint, Alexander Mørch, and Kristina Correa. We thank the technical support of Heather Rada, Kellie Johnson, and Ekaterina Zvezdova (the latter two from BioLegend). We thank Professor Catarina E. Hioe for kindly providing the HIV-1 gp120 protein. We would also like to thank all the anonymized blood donors who contributed to our study. This work was funded by Wellcome Trust Principal Research Fellowship 100262Z/12/Z, the ERC Advanced Grant (SYNECT AdG 670930), and the Kennedy Trust for Rheumatology Research (KTRR) (all three to M.L.D.). P.F.C.D was supported by EMBO Long-Term Fellowship (ALTF 1420–2015, in conjunction with the European Commission (LTFCOFUND2013, GA-2013-609409) and Marie Sklodowska-Curie Actions) and Oxford-Bristol Myers Squibb Fellowship. A.K. was supported by H2020 and the Research Council of Norway (in conjunction with Marie Sklodowska-Curie Actions 275466; to A.K.). M.F. and H.C.Y. thank the Wellcome Trust (212343/Z/18/Z) and EPSRC (EP/S004459/1). The eTIRF-SIM platform was built-in collaboration with Micron (www.micronoxford.com), an Oxford-wide advanced microscopy technology consortium supported by Wellcome Strategic Awards (091911 and 107457), and with additional funds from an MRC/EPSRC/BBSRC next-generation imaging award and the Kennedy Trust for Rheumatology Research through the Kennedy Institute Cell Dynamics Platform. We acknowledge the generous support of the Kennedy Trust for Rheumatology Research, IDRM, and Carl Zeiss GMBH for the Airyscan LSM 980 confocal microscope used in this research. Y.P., T.D., and R.F. were supported by the Chinese Academy of Medical Sciences (CAMS) Innovation Fund for Medical Sciences (CIFMS), China (grant number: 2018-I2M-2-002) and UK Medical Research Council (MRC); E.S. was supported by Newton, The immunological synapse is a molecular hub that facilitates the delivery of three activation signals, namely antigen, costimulation/corepression and cytokines, from antigen-presenting cells (APC) to T cells. T cells release a fourth class of signaling entities, trans-synaptic vesicles (tSV), to mediate bidirectional communication. Here we present bead-supported lipid bilayers (BSLB) as versatile synthetic APCs to capture, characterize and advance the understanding of tSV biogenesis. Specifically, the integration of juxtacrine signals, such as CD40 and antigen, results in the adaptive tailoring and release of tSV, which differ in size, yields and immune receptor cargo compared with steadily released extracellular vesicles (EVs). Focusing on CD40L+ tSV as model effectors, we show that PD-L1 trans-presentation together with TSG101, ADAM10 and CD81 are key in determining CD40L vesicular release. Lastly, we find greater RNA-binding protein and microRNA content in tSV compared with EVs, supporting the specialized role of tSV as intercellular messengers., Depto. de Biología Celular, Fac. de Ciencias Biológicas, TRUE, pub

HLA-G immune modulatory genes and molecules are presently being studied by a widespread number of research groups. In the present study, we do not aim to be exhaustive since the number of manuscripts published every year is overwhelming. Instead, our aim is pointing out facts about HLA-G function, polymorphism and pathology that have been confirmed by several different researchers, together with exposing aspects that may have been overlooked or not sufficiently remarked in this productive field of study. On the other hand, we question whether performing mainly studies on HLA-G and disease associations is going to give a clear answer in the future, since 40 years of study of classical HLA molecules association with disease has still given no definite answer on this issue., FECYT, ISCIII, UCM, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Ultraviolet (UV) irradiation has profound effects on the skin and the systemic immune system. Several effects of UV radiation on Dendritic cells (DCs) functions have been described. However, gene expression changes induced by UV radiation in DCs have not been addressed before. In this report, we irradiated human monocyte-derived DCs with solar-simulated UVA/UVB and analyzed regulated genes on human whole genome arrays. Results were validated by RT-PCR and further analyzed by Gene Set Enrichment Analysis (GSEA). Solar-simulated UV radiation up-regulated expression of genes involved in cellular stress and inflammation, and down-regulated genes involved in chemotaxis, vesicular transport and RNA processing. Twenty four genes were selected for comparison by RT-PCR with similarly treated human primary keratinocytes and human melanocytes. Several genes involved in the regulation of the immune response were differentially regulated in UVA/UVB irradiated human monocyte-derived DCs, such as protein tyrosine phosphatase, receptor type E (PTPRE), thrombospondin-1 (THBS1), inducible costimulator ligand (ICOSL), galectins, Src-like adapter protein (SLA), IL-10 and CCR7. These results indicate that UV-exposure triggers the regulation of a complex gene repertoire involved in human-DC–mediated immune responses., Ministerio de Educación yCiencia, Programa de Estimulo para laTransferencia de Resultados de la Investigacion(PETRI), I.F. Cantabria, Depto. de Biología Celular, Fac. de Ciencias Biológicas, TRUE, pub

Preterm labor (PTL) is the leading cause of neonatal morbidity and mortality worldwide. Whereas many studies have investigated the maternal immune responses that cause PTL, fetal immune cell activation has recently been raised as an important contributor to the pathogenesis of PTL. In this study, we analyzed lymphocyte receptor repertoires in maternal and cord blood from 14 term and 10 preterm deliveries, hypothesizing that the high prevalence of infection in patients with PTL may result in specific changes in the T cell and B cell repertoires. We analyzed TCR b-chain (TCR-b) and IgH diversity, CDR3 lengths, clonal sharing, and preferential usage of variable and joining gene segments. Both TCR-b and IgH repertoires had shorter CDR3s compared with those in maternal blood. In cord blood samples, we found that CDR3 lengths correlated with gestational age, with shorter CDR3s in preterm neonates suggesting a less developed repertoire. Preterm cord blood displayed preferential usage of a number of genes. In preterm pregnancies, we observed significantly higher prevalence of convergent clones between mother/baby pairs than in term pregnancies. Together, our results suggest the repertoire of preterm infants displays a combination of immature features and convergence with maternal TCR-b clones compared with that of term infants. The higher clonal convergence in PTL could represent mother and fetus both responding to a shared stimulus like an infection. These data provide a detailed analysis of the maternaletal immune repertoire in term and preterm patients and contribute to a better understanding of neonate immune repertoire development and potential changes associated with PTL, Burroughs Wellcome Fund, National Institutes of Health, Ulla og Mogens Folmer Andersens Fond, Depto. de Estadística y Ciencia de los Datos, Fac. de Estudios Estadísticos, TRUE, pub

HLA-G is a non-classical class I HLA molecule that induces tolerance by acting on receptors of both innate and adaptive immune cells. When overexpressed in tumors, limits surveillance by the immune system. The HLA-G gene shows several polymorphisms involved in mRNA and protein levels. We decided to study the implication of two polymorphisms (rs371194629; 14bp INS/DEL and rs1063320; +3142 C/G) in paired tissue samples (tumoral and non-tumoral) from 107 Spanish patients with gastric adenocarcinoma and 58 healthy control individuals, to assess the possible association of the HLA-G gene with gastric adenocarcinoma susceptibility, disease progression and survival. The presence of somatic mutations involving these polymorphisms was also analyzed. The frequency of the 14bp DEL allele was increased in patients (70.0%) compared to controls (57.0%, p=0.025). In addition, the haplotype formed by the combination of the 14bp DEL/+3142 C variants is also increased in patients (54.1% vs 44.4%, p=0.034, OR=1.74 CI95% 1.05-2.89). Kaplan-Meier analysis revealed that 14bp DEL/DEL patients showed lower 5-year life-expectancy than INS/DEL or INS/INS (p=0.041). Adjusting for TNM staging (Cox regression analysis) disclosed a significant difference in death risk (p=0.03) with an expected hazard 2.6 times higher. Finally, no somatic mutations were found when comparing these polymorphisms in tumoral vs non-tumoral tissues, which indicates that this is a preexisting condition in patients and not a de novo, tumor-restricted, event. In conclusion, the variants predominant in patients were those increasing HLA-G mRNA stability and HLA-G expression, clearly involving this molecule in gastric adenocarcinoma susceptibility, disease progression and survival and making it a potential target for immunotherapeutic approaches., Unión Europea, Instituto de SaludCarlos III, Universidad Complutense de Madrid, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Instituciones participantes: 1Department of Internal Medicine III, University Hospital, RWTH Aachen; 2Department of Gastroenterology and Hepatology, University Hospital Duisburg-Essen, Essen, Germany; 3Nutrition, Metabolism & Genomics group, Wageningen University, Division of Human Nutrition, Wageningen, The Netherlands; 4Norwich Medical School, University of East Anglia, Norwich, United Kingdom; 5Proteomics Facility, University Hospital, RWTH Aachen; 6Institute of Pathology, University Hospital, RWTH Aachen, Germany; and 7Howard Hughes Medical Institute and University of Massachusetts Medical School, Worcester, Massachusetts, BACKGROUND & AIMS: c-Jun N-terminal kinase (JNK) 1 and JNK2 are expressed in hepatocytes and have overlapping and distinct functions. JNK proteins are activated via phosphorylation in response to acetaminophen- or carbon tetrachloride (CCl4)- induced liver damage; the level of activation correlates with the degree of injury. SP600125, a JNK inhibitor, has been reported to block acetaminophen-induced liver injury. We investigated the role of JNK in drug-induced liver injury (DILI) in liver tissue from patients and in mice with genetic deletion of JNK in hepatocytes. METHODS: We studied liver sections from patients with DILI (due to acetaminophen, phenprocoumon, nonsteroidal antiinflammatory drugs, or autoimmune hepatitis) or patients without acute liver failure (controls) collected from a DILI Biobank in Germany. Levels of total and activated (phosphorylated) JNK were measured by immunohistochemistry and Western blotting. Mice with hepatocyte-specific deletion of Jnk1 (Jnk1Dhepa) or combination of Jnk1 and Jnk2 (JnkDhepa), as well as Jnk1-floxed C57BL/6 (control) mice, were given injections of CCl4 (to induce fibrosis) or acetaminophen (to induce toxic liver injury). We performed gene expression microarray and phosphoproteomic analyses to determine mechanisms of JNK activity in hepatocytes. RESULTS: Liver samples from DILI patients contained more activated JNK, predominantly in nuclei of hepatocytes and in immune cells, than healthy tissue. Administration of acetaminophen to JnkDhepa mice produced a greater level of liver injury than that observed in Jnk1Dhepa or control mice, based on levels of serum markers and microscopic and histologic analysis of liver tissues. Administration of CCl4 also induced stronger hepatic injury in JnkDhepa mice, based on increased inflammation, cell proliferation, and fibrosis progression, compared with Jnk1Dhepa or control mice. Hepatocytes from JnkDhepa mice given acetaminophen had an increased oxidative stress response, leading to de, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Background: Polymerized allergoids conjugated to mannan (PM) are suitable vaccines for allergen-specific immunotherapy (AIT). Alum remains the most widely used adjuvant in AIT, but its way of action is not completely elucidated. The better understanding of the mechanisms underlying alum adjuvanticity could help to improve AIT vaccine formulations. Objective: We sought to investigate the potential influence of alum in the tolerogenic properties imprinted by PM at the molecular level. Methods: Flow cytometry, ELISAs, cocultures, intracellular staining and suppression assays were performed to assess alum and PM effects in human dendritic cells (DCs). BALB/c mice were immunized with PM alone or adsorbed to alum. Allergen-specific antibodies, splenocyte cytokine production and splenic forkhead box P3 (FOXP3)+ regulatory T (Treg) cells were quantified. Metabolic and immune pathways were also studied in human DCs. Results: Alum decreases PD-L1 expression and IL-10 production induced by PM in human DCs and increases pro-inflammatory cytokine production. Alum impairs PM-induced functional FOXP3+ Treg cells and promotes Th1/Th2/Th17 responses. Subcutaneous immunization of mice with PM plus alum inhibits in vivo induction of Treg cells promoted by PM without altering the capacity to induce functional allergen-specific blocking antibodies. Alum inhibits mTOR activation and alters metabolic reprogramming by shifting glycolytic pathways and inhibiting reactive oxygen species (ROS) production in PM-activated DCs, impairing their capacity to generate functional Treg cells. Conclusion: We uncover novel mechanisms by which alum impairs the tolerogenic properties induced by PM, which might well contribute to improve the formulation of novel vaccines for AIT., Ministerio de Economía y Competitividad, Centre for Industrial Technological Development, Depto. de Bioquímica y Biología Molecular, Fac. de Ciencias Químicas, TRUE, pub

Acetaminophen (APAP) hepatotoxicity induces endoplasmic reticulum (ER) stress which triggers the unfolded protein response (UPR) in hepatocytes. However, the mechanisms underlying ER stress remain poorly understood, thus reducing the options for exploring new pharmacological therapies for patients with hyperacute liver injury. Eight-to-twelve-week-old C57BL/6J Xbp1-floxed (Xbp1f/f) and hepatocyte-specific knockout Xbp1 mice (Xbp1∆hepa) were challenged with either high dose APAP [500 mg/kg] and sacrificed at early (1-2 h) and late (24 h) stages of hepatotoxicity. Histopathological examination of livers, immunofluorescence and immunohistochemistry, Western blot, real time (RT)-qPCR studies and transmission electron microscopy (TEM) were performed. Pharmacological inhibition of XBP1 using pre-treatment with STF-083010 [STF, 75 mg/kg] and autophagy induction with Rapamycin [RAPA, 8 mg/kg] or blockade with Chloroquine [CQ, 60 mg/kg] was also undertaken in vivo. Cytoplasmic expression of XBP1 coincided with severity of human and murine hyperacute liver injury. Transcriptional and translational activation of the UPR and sustained activation of JNK1/2 were major events in APAP hepatotoxicity, both in a human hepatocytic cell line and in a preclinical model. Xbp1∆hepa livers showed decreased UPR and JNK1/2 activation but enhanced autophagy in response to high dose APAP. Additionally, blockade of XBP1 splicing by STF, mitigated APAP-induced liver injury and without non-specific off-target effects (e.g., CYP2E1 activity). Furthermore, enhanced autophagy might be responsible for modulating CYP2E1 activity in Xbp1∆hepa animals. Genetic and pharmacological inhibition of Xbp1 specifically in hepatocytes ameliorated APAP-induced liver injury by enhancing autophagy and decreasing CYP2E1 expression. These findings provide the basis for the therapeutic restoration of ER stress and/or induction of autophagy in patients with hyperacute liver injury., Ministerio de Economía, Comercio y Empresa, German Research Foundation, Ministerio de Ciencia, Innovación y Universidades, Fundación Científica de la Asociación Española Contra el Cancer, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Department of Medicine. Division of Liver Diseases. Mount Sinai School of Medicine. New York, USA, Liver disease is a major cause of illness and death worldwide. A central component in the complex network leading to the development of alcoholic liver disease is the activation of Kupffer cells by endotoxin and other soluble mediators. Alcohol consumption induces a state of "leaky gut increasing plasma and liver endotoxin levels. When Kupffer cells become activated, they interact with a complex of proteins located on the extracellular membrane signaling to produce a wide array of soluble factors, including cytokines, chemokines, growth factors, cyclooxygenase and lipoxygenase metabolites, and reactive oxygen species such as superoxide anion, hydrogen peroxide, and nitric oxide, all of which provide physiologically diverse and pivotal paracrine effects on all other liver cell types and, ultimately, liver injury. Kupffer cells are also central to the liver homeostatic response to injury as upon cellular degenerative changes, they immediately respond to the insult and release mediators to orchestrate inflammatory and reparative responses. Thus, the homeostatic responses are initiated by Kupffer cell-derived mediators at the cellular level and underlie the liver s defense and reparative mechanisms against injury. In order to understand better the role of Kupffer cells in the onset of liver injury, animal models in which Kupffer cells are inactivated, and cell culture settings (e.g. co-cultures) are being used with promising results that advance our understanding of alcoholic liver disease., National Institute of Diabetes and Digestive and Kidney Diseases (US Public Health Service), Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Extracellular vesicles are potent modulators of immunity. This Research Topic provided a platform for organizing the recent findings and future perspectives on EV’s biology and functions in various immune-related processes. We hope that he published results and conclusions will become an impulse for new research discoveries to overcome some of the current drawbacks for EV-based therapies in a way that will determine the future directions of clinical practice., Depto. de Biología Celular, Fac. de Ciencias Biológicas, TRUE, pub

La estimulación de células presentadoras de antígeno con LPS de Porphyromonas gingivalis da lugar a la producción de ciertas citocinas proinflamatorias y antiinflamatorias. Sin embargo, las vías de señalización que regulan estos procesos no son del todo conocidas. En el presente estudio se investigó el papel de la vía fosfatidilinositol 3 quinasa (PI3K)-Akt en la regulación de la producción de IL-10, IL-12 p40 e IL-12 p70 inducida por LPS de P. gingivalis en monocitos humanos. El LPS de P. gingivalis activó selectivamente la vía PI3K-Akt a través del receptor TLR2, y la inhibición de esta vía dio lugar a una abrogación de la fosforilación de ERK1/2, mientras que la activación de las quinasas p38 y JNK 1/2 no se vio afectada. El análisis de la producción de citoquinas tras la estimulación de monocitos con LPS de P. gingivalis reveló que la inhibición de la vía PI3K regulaba diferencialmente la síntesis de IL-10 e IL-12. Se suprimía la producción de IL-10, mientras que los niveles de IL-12 aumentaban. La inhibición de la activación de la vía PI3K-Akt dio lugar a un aumento pronunciado de NF-kB p65 que era independiente de la degradación de IkB-alfa. Además, la capacidad de la vía PI3K-Akt para modular la producción de IL-10 e IL-12 parece estar mediada por la supresión selectiva de la actividad de ERK1/2, ya que el tratamiento con el inhibidor de MEK1 PD98059 producía efectos similares a la wortmanina y LY294002, regulando diferencialmente la producción de IL-10 e IL-12 en monocitos estimulados por LPS de P gingivalis. Estos estudios proporcionan una nueva visión de cómo el acoplamiento de la vía PI3K-Akt por parte del LPS de P. gingivalis afecta a la inducción de citocinas inmunorreguladoras clave que controlan tanto los aspectos cualitativos como cuantitativos de la inmunidad innata y adaptativa., Depto. de Biología Celular, Fac. de Medicina, TRUE, pub

Los receptores tipo Toll (TLR) median la activación celular por parte de los microbios y los productos microbianos. Para estudiar el papel de los TLR en el desarrollo de las respuestas inmunitarias del huésped contra las micobacterias, se infectaron ratones de tipo salvaje y deficientes en TLR2 o TLR4 con el bacilo no patógeno Mycobacterium bovis Calmette-Guerin (BCG). Dos semanas después de la exposición intraperitoneal con BCG, había pocos bacilos presentes en los pulmones de los ratones de tipo salvaje y TLR4(-/-), mientras que las cargas bacterianas eran diez veces mayores en los pulmones de los ratones TLR2(-/-) infectados. El desafío con BCG in vitro indujo fuertemente la secreción de citoquinas proinflamatorias por parte de los macrófagos de los ratones de tipo salvaje y TLR4 (-/-), pero no por los macrófagos TLR2 (-/-). Por el contrario, el transporte intracelular, el crecimiento bacteriano intracelular y la supresión del crecimiento bacteriano intracelular in vitro por interferón-gamma (IFN-gamma) fueron similares en los macrófagos de las tres cepas de ratón, lo que sugiere que el crecimiento de BCG en los pulmones de los ratones TLR2 (-/-) fue consecuencia de una inmunidad adaptativa defectuosa. La estimulación antigénica de los esplenocitos de ratones infectados de tipo salvaje y TLR4(-/-) indujo la proliferación de células T in vitro, mientras que las células T de ratones TLR2(-/-) no lograron proliferar. Inesperadamente, las células T CD4(+) activadas de ambas cepas de ratón deficientes en TLR secretaron poco IFN-gamma in vitro en comparación con las células T de los ratones control. El papel de TLR4 en el control del crecimiento bacteriano y la producción de IFN-gamma in vivo se observó solo cuando los ratones estaban infectados con un mayor número de BCG. Por lo tanto, TLR2 y TLR4 parecen regular distintos aspectos de la respuesta inmune del huésped contra BCG., Depto. de Biología Celular, Fac. de Medicina, TRUE, pub

The SARS-CoV-2 pandemic has a huge impact on public health and global economy, meaning an enormous scientific, political, and social challenge. Studying how infection or vaccination triggers both cellular and humoral responses is essential to know the grade and length of protection generated in the population. Nowadays, scientists and authorities around the world are increasingly concerned about the arrival of new variants, which have a greater spread, due to the high mutation rate of this virus. The aim of this review is to summarize the different techniques available for the study of the immune responses after exposure or vaccination against SARS-CoV-2, showing their advantages and limitations, and proposing suitable combinations of different techniques to achieve extensive information in these studies. We wish that the information provided here will helps other scientists in their studies of the immune response against SARS-CoV-2 after vaccination with new vaccine candidates or infection with upcoming variants., Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Cholestatic liver diseases including primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) are associated with active hepatic fibrogenesis, which can ultimately lead to the development of cirrhosis. However, the exact relationship between the development of liver fibrosis and the progression of cholestatic liver disease remains elusive. Periductular fibroblasts located around the bile ducts seem biologically different from hepatic stellate cells (HSCs). The fibrotic events in these clinical conditions appear to be related to complex crosstalk between immune/inflammatory mechanisms, cytokine signalling, and perturbed homeostasis between cholangiocytes and mesenchymal cells. Several animal models including bile duct ligation (BDL) and the Mdr2-knockout mice have improved our understanding of mechanisms underlying chronic cholestasis. In the present review, we aim to elucidate the mechanisms of fibrosis in order to help to identify potential diagnostic and therapeutic targets., Cholestatic liver diseases including primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) are associated with active hepatic fibrogenesis, which can ultimately lead to the development of cirrhosis. However, the exact relationship between the development of liver fibrosis and the progression of cholestatic liver disease remains elusive. Periductular fibroblasts located around the bile ducts seem biologically different from hepatic stellate cells (HSCs). The fibrotic events in these clinical conditions appear to be related to complex crosstalk between immune/inflammatory mechanisms, cytokine signalling, and perturbed homeostasis between cholangiocytes and mesenchymal cells. Several animal models including bile duct ligation (BDL) and the Mdr2knockout mice have improved our understanding of mechanisms underlying chronic cholestasis. In the present review, we aim to elucidate the mechanisms of fibrosis in order to help to identify potential diagnostic and therapeutic targets., Ministerio de Economía, Comercio y Empresa, German Research Foundation, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Background: Infiltrating B and T cells have been observed in several tumor tissues, including pancreatic ductal adenocarcinoma (PDAC). The majority known PDAC risk factors point to a chronic inflammatory process leading to different forms of immunological infiltration. Understanding pancreatic tumor infiltration may lead to improved knowledge of this devastating disease. Methods: We extracted the immunoglobulins (IGs) and T cell receptors (TCRs) from RNA-sequencing of 144 PDAC from TCGA and 180 pancreatic normal tissue from GTEx. We used Shannon entropy to find differences in IG/TCR diversity. We performed a clonotype analysis considering the IG clone definition (same V and J segments, same CDR3 length, and 90% nucleotide identity between CDR3s) to study differences among the tumor samples. Finally, we performed an association analysis to find host and tumor factors associated with the IG/TCR. Results: PDAC presented a richer and more diverse IG and TCR infiltration than normal pancreatic tissue. A higher IG infiltration was present in heavy smokers and females and it was associated with better overall survival. In addition, specific IG clonotypes classified samples with better prognosis explaining 24% of the prognosis phenotypic variance. On the other hand, a larger TCR infiltration was present in patients with previous history of diabetes and was associated with lower nonantigen load. Conclusions: Our findings support PDAC subtyping according to its immune repertoire landscape with a potential impact on the understanding of the inflammatory basis of PDAC risk factors as well as the design of treatment options and prognosis monitoring, AACRAstraZeneca, Fondo de InvestigacionesSanitarias (FIS), Instituto de Salud Carlos III, Spain, Pancreatic Cancer Collective (PCC), Lustgarten Foundation & Stand-Up to Cancer, Depto. de Estadística y Ciencia de los Datos, Fac. de Estudios Estadísticos, TRUE, pub

Background: Experimental autoimmune myocarditis (EAM), a mouse model of post-infectious cardiomyopathy, reflects mechanisms of inflammatory cardiomyopathy in humans. EAM is characterized by an infiltration of inflammatory cells into the myocardium that can be followed by myocyte fibrosis, edema, and necrosis, leading to ventricular wall dysfunction and heart failure. Different data indicate that CD69 exerts an important immunoregulatory effect in vivo. However, the possible role of CD69 in autoimmune myocarditis has not been studied. Methods and results: We have explored the role of the leukocyte regulatory molecule CD69 in the inflammation that leads to cardiac dysfunction after myocardial injury in EAM. We have found that after induction of EAM, the draining lymph nodes from CD69-deficient mice developed an exacerbated Th17 inflammatory response, resulting in increases in the numbers of infiltrating leukocytes in the myocardium. In the chronic phase of EAM, transthoracic echocardiography revealed a significantly reduced left ventricular fractional shortening and a decreased ejection fraction in CD69-deficient mice, indicative of an impaired cardiac contractility. This condition was accompanied by a greater extent of myocardial fibrosis, an elevated number of sinus pauses on ECG, and an enhanced ratio of heart weight to body weight in CD69-/- mice. Moreover, both bone marrow transplantation and adoptive transfer of Th17 cells isolated from immunized CD69-/- mice with EAM into naive wild-type recipients reproduced the severity of the disease, demonstrating that CD69 exerts its function within the lymphocyte compartment. Conclusion: Our findings indicate that CD69 negatively regulates heart-specific Th17 responses, cardiac inflammation, and heart failure progression in EAM., Ministerio de Ciencia e Innovacion, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Regulatory T cells (Tregs) control immune responses and are essential to maintain immune homeostasis and self-tolerance. Hence, it is no coincidence that autoimmune and chronic inflammatory disorders are associated with defects in Tregs. These diseases have currently no cure and are treated with palliative drugs such as immunosuppressant and immunomodulatory agents. Thereby, there is a great interest in developing medical interventions against these diseases based on enhancing Treg cell function and numbers. Here, we give an overview of Treg cell ontogeny and function, paying particular attention to mucosal Tregs. We review some notable approaches to enhance immunomodulation by Tregs with therapeutic purposes including adoptive Treg cell transfer therapy and discuss relevant clinical trials for inflammatory bowel disease. We next introduce ways to expand mucosal Tregs in vivo using microbiota and dietary products that have been the focus of clinical trials in various autoimmune and chronic-inflammatory diseases., Comunidad Autonoma de Madrid, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Respiratory disorders caused by allergy have been associated to bronchiolar inflammation leading to life-threatening airway narrowing. However, whether airway allergy causes alveolar dysfunction contributing to the pathology of allergic asthma remains unaddressed. To explore whether airway allergy causes alveolar dysfunction that might contribute to the pathology of allergic asthma, alveolar structural and functional alterations were analyzed during house dust mite (HDM)-induced airway allergy in mice, by flow cytometry, light and electron microscopy, monocyte transfer experiments, assessment of intra-alveolarly-located cells, analysis of alveolar macrophage regeneration in Cx3cr1cre:R26-yfp chimeras, analysis of surfactant-associated proteins, and study of lung surfactant biophysical properties by captive bubble surfactometry. Our results demonstrate that HDM-induced airway allergic reactions caused severe alveolar dysfunction, leading to alveolar macrophage death, pneumocyte hypertrophy and surfactant dysfunction. SP-B/C proteins were reduced in allergic lung surfactant, that displayed a reduced efficiency to form surface-active films, increasing the risk of atelectasis. Original alveolar macrophages were replaced by monocyte-derived alveolar macrophages, that persisted at least two months after the resolution of allergy. Monocyte to alveolar macrophage transition occurred through an intermediate stage of pre-alveolar macrophage and was paralleled with translocation into the alveolar space, Siglec-F upregulation, and downregulation of CX3CR1. These data support that the severe respiratory disorders caused by asthmatic reactions not only result from bronchiolar inflammation, but additionally from alveolar dysfunction compromising an efficient gas exchange., Ministerio de Economía y Competitividad, Ministerio de Ciencia e Innovación, Comunidad de Madrid, Depto. de Bioquímica y Biología Molecular, Fac. de Ciencias Químicas, TRUE, pub

Type 3 Innate lymphoid cells (ILC3s) have been described as tissue-resident cells and characterized throughout the body, especially in mucosal sites and classical first barrier organs such as skin, gut and lungs, among others. A significant part of the research has focused on their role in combating pathogens, mainly extracellular pathogens, with the gut as the principal organ. However, some recent discoveries in the field have unveiled their activity in other organs, combating intracellular pathogens and as part of the response to viruses. In this review we have compiled the latest studies on the role of ILC3s and the molecular mechanisms involved in defending against different microbes at the mucosal surface, most of these studies have made use of conditional transgenic mice. The present review therefore attempts to provide an overview of the function of ILC3s in infections throughout the body, focusing on their specific activity in different organs., Programa Ramón y Cajal, Ministerio de Ciencia, Innovación e Universidades, Fondo Europeo de Desarrollo Regional, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Department of Medicine, Division of Liver Diseases, Mount Sinai School of Medicine, New York, New York, Background & Aims: Liver injury leads to generation of reactive oxygen and nitrogen species, which can react to produce peroxynitrite (ONOO). We investigated whether ONOO and its metabolites modulate extracellular matrix remodeling. Methods: Stellate cells (HSC) were incubated with pure ONOO or SIN-1 (a ONOO donor). Western blot, nuclear in vitro transcription, Northern blot, qPCR, and promoter transactivation analysis for COL1A1 and COL1A2 were carried out. Rats were fed alcohol or injected with CCl4 to cause alcohol-induced liver injury and an early fibrogenic response. Results: HSC incubated with ONOO or SIN-1 showed similar viability, proliferation, COL1A1 and COL1A2 transcription rates, and mRNA levels as controls. There was a time- and dose-dependent down-regulation of collagen I and alfa-Sma proteins and up-regulation of MMP1 and TNF , indicating decreased HSC activation. These effects were blocked by ONOO scavengers. SIN-1 or ONOO increased nitrosylation of MMP1/MMP13 and transactivation of the MMP1, MMP13, and TNFalfa promoters. A TNFalfa neutralizing antibody or GSH-ethyl ester blocked MMP1 promoter transactivation; whereas TNFalfa or L-buthionine sulfoximine, which depletes GSH, further enhanced it. Pretreatment with SIN-1 or ONOO- reduced the TGF pro-fibrogenic response in HSC. In vivo experiments validated the protective role of ONOO- on the early fibrogenic response. However, highly activated HSC, such as myofibroblasts and HSC from chronic alcohol-fed rats, were resistant to the anti-fibrogenic actions of ONOO- due to higher levels of GSH, a ONOO- scavenger, overproduction of pro-fibrogenic TGF, and reactive oxygen species. Conclusion: ONOO- could induce a protective mechanism in HSC in early stages of liver injury., National Institute of Diabetes and Digestive and Kidney Diseases (US Public Health Service), Gobierno de Navarra, Ministerio de Educación y Ciencia, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Purpose of Review: Allergic diseases represent a major health problem of increasing prevalence worldwide. In allergy, dendritic cells (DCs) contribute to both the pathophysiology and the induction of healthy immune responses to the allergens. Different studies have reported that some common allergens contain glycans in their structure. C-type lectin receptors (CLRs) expressed by DCs recognize carbohydrate structures and are crucial in allergen uptake, presentation, and polarization of T cell responses. This review summarizes the recent literature regarding the role of CLRs in the regulation of type 2 immune responses to allergens. Recent Findings: In this review, we highlight the capacity of CLRs to recognize carbohydrates in common allergens triggering different signaling pathways involved in the polarization of CD4+ T cells towards specific Th2 responses. Under certain conditions, specific CLRs could also promote tolerogenic responses to allergens, which might well be exploited to develop novel therapeutic approaches of allergen-specific immunotherapy (AIT), the single treatment with potential disease-modifying capacity for allergic disease. At this regard, polymerized allergens conjugated to non-oxidized mannan (allergoid-mannan conjugated) are next-generation vaccines targeting DCs via CLRs that promote regulatory T cells, thus favoring allergen tolerance both in preclinical models and clinical trials. Summary: A better understanding of the role of CLRs in the development of allergy and in the induction of allergen tolerance might well pave the way for the design of novel strategies for allergic diseases., MICINN, Depto. de Bioquímica y Biología Molecular, Fac. de Ciencias Químicas, TRUE, pub

Supplementary Information accompanies this paper at https://doi.org/10.1038/s41467-019-09930-3. Competing interests: The authors declare no competing interests. Reprints and permission information is available online at http://npg.nature.com/reprintsandpermissions/ Journal peer review information: Nature Communications thanks the anonymousreviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available. Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations., Studying immune repertoire in the context of organ transplant provides important information on how adaptive immunity may contribute and modulate graft rejection. Here we characterize the peripheral blood immune repertoire of individuals before and after kidney transplant using B cell receptor sequencing in a longitudinal clinical study. Individuals who develop rejection after transplantation have a more diverse immune repertoire before transplant, suggesting a predisposition for post-transplant rejection risk. Additionally, over 2 years of follow-up, patients who develop rejection demonstrate a specific set of expanded clones that persist after the rejection. While there is an overall reduction of peripheral B cell diversity, likely due to increased general immunosuppression exposure in this cohort, the detection of specific IGHV gene usage across all rejecting patients supports that a common pool of immunogenic antigens may drive post-transplant rejection. Our findings may have clinical implications for the prediction and clinical management of kidney transplant rejection., Depto. de Estadística y Ciencia de los Datos, Fac. de Estudios Estadísticos, TRUE, pub