African swine fever virus infection in Classical swine fever subclinically infected wild boars

Recently moderate-virulence classical swine fever virus (CSFV) strains have been proven capable of generating postnatal persistent infection (PI), defined by the maintenance of viremia and the inability to generate CSFV-specific immune responses in animals. These animals also showed a type I interferon blockade in the absence of clinical signs. In this study, we assessed the infection generated in 7-week-old CSFV PI wild boars after infection with the African swine fever virus (ASFV). The wild boars were divided in two groups and were infected with ASFV. Group A comprised boars who were CSFV PI in a subclinical form and Group B comprised pestivirus-free wild boars. Some relevant parameters related to CSFV replication and the immune response of CSFV PI animals were studied. Additionally, serum soluble factors such as IFN-α, TNF-α, IL-6, IL-10, IFN-γ and sCD163 were analysed before and after ASFV infection to assess their role in disease progression.This research was supported by grants AGL2013–48998 and AGL2015–66907 from the Spanish government. S.M and A.C ha

Development of a framework for fire risk assessment using remote sensing and geographic information system technologies

Forest fires play a critical role in landscape transformation, vegetation succession, soil degradation and air quality. Improvements in fire risk estimation are vital to reduce the negative impacts of fire, either by lessen burn severity or intensity through fuel management, or by aiding the natural vegetation recovery using post-fire treatments. This paper presents the methods to generate the input variables and the risk integration developed within the Firemap project (funded under the Spanish Ministry of Science and Technology) to map wildland fire risk for several regions of Spain. After defining the conceptual scheme for fire risk assessment, the paper describes the methods used to generate the risk parameters, and presents proposals for their integration into synthetic risk indices. The generation of the input variables was based on an extensive use of geographic information system and remote sensing technologies, since the project was intended to provide a spatial and temporal assessment of risk conditions. All variables were mapped at 1 km2 spatial resolution, and were integrated into a web-mapping service system. This service was active in the summer of 2007 for semi-operational testing of end-users. The paper also presents the first validation results of the danger index, by comparing temporal trends of different danger components and fire occurrence in the different study regions.The Firemap project was funded by the Spanish Ministry of Science and Education (CGL2004-060490C04-01/CLI) through the Environment and Climate programPeer reviewe

Peri-operative red blood cell transfusion in neonates and infants: NEonate and Children audiT of Anaesthesia pRactice IN Europe: A prospective European multicentre observational study

BACKGROUND: Little is known about current clinical practice concerning peri-operative red blood cell transfusion in neonates and small infants. Guidelines suggest transfusions based on haemoglobin thresholds ranging from 8.5 to 12 g dl-1, distinguishing between children from birth to day 7 (week 1), from day 8 to day 14 (week 2) or from day 15 (≥week 3) onwards. OBJECTIVE: To observe peri-operative red blood cell transfusion practice according to guidelines in relation to patient outcome. DESIGN: A multicentre observational study. SETTING: The NEonate-Children sTudy of Anaesthesia pRactice IN Europe (NECTARINE) trial recruited patients up to 60 weeks' postmenstrual age undergoing anaesthesia for surgical or diagnostic procedures from 165 centres in 31 European countries between March 2016 and January 2017. PATIENTS: The data included 5609 patients undergoing 6542 procedures. Inclusion criteria was a peri-operative red blood cell transfusion. MAIN OUTCOME MEASURES: The primary endpoint was the haemoglobin level triggering a transfusion for neonates in week 1, week 2 and week 3. Secondary endpoints were transfusion volumes, 'delta haemoglobin' (preprocedure - transfusion-triggering) and 30-day and 90-day morbidity and mortality. RESULTS: Peri-operative red blood cell transfusions were recorded during 447 procedures (6.9%). The median haemoglobin levels triggering a transfusion were 9.6 [IQR 8.7 to 10.9] g dl-1 for neonates in week 1, 9.6 [7.7 to 10.4] g dl-1 in week 2 and 8.0 [7.3 to 9.0] g dl-1 in week 3. The median transfusion volume was 17.1 [11.1 to 26.4] ml kg-1 with a median delta haemoglobin of 1.8 [0.0 to 3.6] g dl-1. Thirty-day morbidity was 47.8% with an overall mortality of 11.3%. CONCLUSIONS: Results indicate lower transfusion-triggering haemoglobin thresholds in clinical practice than suggested by current guidelines. The high morbidity and mortality of this NECTARINE sub-cohort calls for investigative action and evidence-based guidelines addressing peri-operative red blood cell transfusions strategies. TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT02350348

A first update on mapping the human genetic architecture of COVID-19

peer reviewe

CD200R family receptors are expressed on porcine monocytes and modulate the production of IL-8 and TNF-α triggered by TLR4 or TLR7 in these cells

The inhibitory receptor CD200R1 and its paired activating receptor CD200R1L are involved in the regulation of myeloid cell immune responses. The aim of this study was to analyze their distribution, regulation by cytokines, and function in porcine monocyte subsets. We had previously observed that CD200R1 and CD200R1L genes can generate different protein isoforms through alternative mRNA splicing, therefore in this study, we explored the diversity of transcripts in monocyte subsets, and described several new splicing variants of both CD200R1 and CD200R1L, some of which could be expressed on the porcine monocyte surface. A substantial proportion of CD163- SLAII+ and most CD163+SLAII+ monocytes expressed CD200R1 and CD200R1L receptors, while CD163- SLAII- monocytes did not. CD200R1 and CD200R1L expression was down-regulated in monocytes polarized by IFN-ɣ, a cytokine that induces classical activation of macrophages, while IL-10 which gives rise to regulatory macrophages, increased the expression of CD200R1. Finally, treatment of monocyte subsets with a monoclonal antibody specific for the inhibitory CD200R1 receptor and its splicing variants enhanced TNFα and IL-8 production, induced by TLR4 or TLR7 stimulation, suggesting a modulatory role for these receptors on porcine monocyte functions

CD200R1 and CD200R1L expression is regulated during B cell development in swine and modulates the Ig production in response to the TLR7 ligand imiquimoid

The CD200R family comprises a group of paired receptors that can modulate the activation of immune cells. They are expressed both on myeloid cells and lymphocyte subsets. Here we report that the expression of these receptors on porcine B cells is tightly regulated, being mainly expressed on mature cells. The expression of the inhibitory receptors CD200R1 and/or its splicing variant CD200R1X2, either in combination or not with the activating receptor CD200R1L, is upregulated in sIgM+ effector/memory cells, and tends to decline thereafter as these cells progress to plasmablasts or switch the Ig isotype. sIgM+ naïve and primed cells only express, by contrast, the CD200R1X2 receptor. B-1 like cells also express CD200R1 isoforms, either alone or in combination with CD200R1L. Treatment of peripheral blood mononuclear cells with a monoclonal antibody specific for inhibitory receptors, enhances the IgM and IgG production induced by TLR7 stimulation suggesting a modulatory role of B cell functions of these receptors.Funding: This work was supported by grants RTA2014-00003-00-00 from INIA and PID2019-10985RB-I00 from Agencia Estatal de Innovación (Ministerio de Ciencia e Innovación of Spain).Peer reviewe

Expression of TLR4 in swine as assessed by a newly developed monoclonal antibody

Toll-like receptors (TLRs) constitute an ancient family of pattern recognition receptors for conserved microbial structures that allow rapid detection of invading pathogens, triggering immune responses. TLR4 binds lipopolysaccharides (LPS) being involved in the recognition of Gram-negative bacteria. Herein we describe the generation and characterisation of a monoclonal antibody, named 3H3, against porcine TLR4. Its specificity was confirmed by reactivity with TLR4 expressing CHO cell transfectants. On peripheral blood leukocytes TLR4 was preferentially expressed on myelomonocytic cells, with monocytes expressing higher levels than granulocytes. Staining of lung tissue sections showed that TLR4 is also expressed on epithelial cells lining the bronchial tract, a distribution consistent with a surveillance function of bacterial invasion. © 2013 Elsevier B.V

Expression of Siglec-1, -3, -5 and -10 in porcine cDC1 and cDC2 subsets from blood, spleen and lymph nodes and functional capabilities of these cells

8 Pág.Dendritic cells are professional antigen-presenting cells that play a critical role in the development of immune responses. DCs express a variety of Siglecs on their surface, which play a regulatory role modulating their activation through interaction with sialylated structures expressed by cells or pathogens. Here, we characterized the phenotype of porcine conventional dendritic cells subsets from blood, spleen and lymph nodes, emphasizing the analysis of the expression of Siglecs. Siglec-1 was detected in type 1 cDC and, at lower levels, in type 2 cDC in the spleen, being low to negative in blood and lymph node cDC. Siglec-3 and Siglec-5 were expressed in cDC1 at lower levels than in cDC2. Porcine cDCs did not express Siglec-10. cDC2 showed a higher capacity to phagocytose microspheres and to process DQ™-OVA than cDC1, but none of these functions was affected by engagement of Siglec-3 and -5 with antibodies on blood cDC.This work was supported by grant RTA2014-00003-00-00 from INIA.Peer reviewe

Porcine CLEC12B is expressed on alveolar macrophages and blood dendritic cells

10 Pág.CLEC12B is a C-type lectin-like receptor expressed on myeloid cells. In this study, we have characterized the porcine homologue of CLEC12B (poCLEC12B). To this end, we have generated constructs encoding a c-myc tagged version of the whole receptor, or its ectodomain fused to the Fc portion of human IgG1, from a cDNA clone obtained from an alveolar macrophage library, and raised monoclonal antibodies (mAb) against this molecule. Using these mAbs, poCLEC12B was found to be expressed on alveolar macrophages and, at lower levels, on blood conventional type 1 dendritic cells (cDC1) and plasmacytoid DCs. No binding was detected on monocytes, monocyte-derived macrophages or monocyte-derived DCs. Engagement of CLEC12B on alveolar macrophages with mAbs had no apparent effect on cytokine production (TNF-α, IL-8) induced by LPS. These results provide the basis for future investigations aimed to assess the role of poCLEC12B in different microbial infections and to evaluate its potential in vaccination strategies targeting DCs.This work was supported by grant AGL2015-66187-P from Ministry of Economy, Industry and Competitiveness of Spain.Peer reviewe