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Seismic inversion is applied to generate physical property models (P-wave velocity and numerical attenuation) for four profiles in the Nechako-Chilcotin plateau region of south-central British Columbia, Canada. A newly developed method that combines three-dimensional (3-D) travel-time inversion and 2.5-dimensional (2.5-D) visco-acoustic full-waveform inversion was applied to generate the geophysical models from vibroseis data acquired along the preexisting crooked roads. These models are useful for the characterization of rock types in terms of their positions and thicknesses, which may be used in conjunction with geological ground truth to infer the extent of lithostratigraphic units in the subsurface. The velocity structures also may be used for future reprocessing of the seismic reflection data to derive improved images based on the better near-surface velocity models. The subsurface geology of the Nechako-Chilcotin plateau region is complex, resulting from multiple stages of tectonic compression and extension, contemporaneous with the deposition of sediments and volcanic material. Several basin structures are identified from the joint interpretation of the waveform tomography velocity models and post-stack time migration images. The combination of these results enables the extrapolation and characterization of geological structures to ~3 km depth, particularly within the Cenozoic volcanic units that dominate near-surface stratigraphy. Based on the seismic profiles, a fence-diagram geological interpretation that extends to ~3 km depth illustrates the complex structure of the Jurassic to Neogene stratigraphic sequence. Des modeles de proprietes physiques (vitesse des ondes P et attenuation numerique) sont generes par inversion sismique le long de quatre profiles dans la region du plateau de Nechako-Chilcotin au sud-est de la Colombie Britannique. Une methode recemment developpee combine l'inversion tridimensionnelle (3-D) des temps de trajet et une inversion 2.5dimensionnelle (2.5-D) des formes d'ondes viscoacoustiques. Cette methode est utilisee pour generer les modeles geophysiques a partir de donnees sismiques acquises a partir d'une source vibroseis le long de routes curvilignes. De tels modeles sont utiles afin de caracteriser la position et l'epaisseur des successions rocheuses; ainsi, combines a des donnees geologiques de surfaces, ces modeles permettent de deduire l'etendue des unites lithostratigraphiques en sous-surface. Les structures imagees par les modeles de vitesse peuvent aussi etre utilisees pour le retraitement de donnees de sismique reflexion: elles permettent d'obtenir des profils ameliores fondes sur des meilleurs modeles de vitesse en proche surface. La geologie du plateau de Nechako-Chilcotin est complexe, et resulte d'evenements tectoniques successifs de compression et extension durant le depot de materiel sedimentaire et volcanique. Plusieurs bassins sont identifies a partir de l'interpretation conjointe de modeles tomographiques et d'images de migration en temps. La combinaison de ces resultats permet d'extrapoler et de caracteriser les structures geologiques jusqu'a 3 km de profondeur, particulierement au sein des unites volcaniques d'age Cenozoique qui dominent la stratigraphie de proche surface. Une section geologique interpretative entre 0 et 3 km de profondeur est construite a partir des profils sismiques et illustre la complexite structurale de la succession d'age Jurassique a Neogene., Introduction Interest in the Mesozoic (mainly Cretaceous) sedimentary units of the Nechako-Chilcotin plateau has been economically motivated by the potential for hydrocarbon development (Hannigan et al. 1994; Hayes et al. [...]

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Infection with Leptospira spp. is common in Réunion, a tropical island in the Indian Ocean. However, respiratory coinfections between strains of Leptospira spp. and other microorganisms are rarely described. Here, we describe the first reported case of coinfection between Leptospira spp. and Chlamydia pneumoniae, responsible for refractory acute respiratory distress syndrome requiring extracorporeal membrane oxygenation with a favorable outcome. In a case of leptospirosis with severe respiratory illness, testing for respiratory coinfection, especially with atypical pathogens, could explain the seriousness of the clinical condition and lead to specific treatment.

Porcine reproductive and respiratory syndrome (PRRS) has an extensive impact on pig production. The causative virus (PRRSV) is divided into two species, PRRSV-1 (European origin) and PRRSV-2 (North American origin). Within PRRSV-1, PRRSV-1.3 strains, such as Lena, are more pathogenic than PRRSV-1.1 strains, such as Flanders 13 (FL13). To date, the molecular interactions of PRRSV with primary lung mononuclear phagocyte (MNP) subtypes, including conventional dendritic cells types 1 (cDC1) and 2 (cDC2), monocyte-derived DCs (moDC), and pulmonary intravascular macrophages (PIM), have not been thoroughly investigated. Here, we analyze the transcriptome profiles of in vivo FL13-infected parenchymal MNP subpopulations and of in vitro FL13- and Lena-infected parenchymal MNP. The cell-specific expression profiles of in vivo sorted cells correlated with their murine counterparts (AM, cDC1, cDC2, moDC) with the exception of PIM. Both in vivo and in vitro , FL13 infection altered the expression of a low number of host genes, and in vitro infection with Lena confirmed the higher ability of this strain to modulate host response. Machine learning (ML) and gene set enrichment analysis (GSEA) unraveled additional relevant genes and pathways modulated by FL13 infection that were not identified by conventional analyses. GSEA increased the cellular pathways enriched in the FL13 data set, but ML allowed a more complete comprehension of functional profiles during FL13 in vitro infection. Data indicates that cellular reprogramming differs upon Lena and FL13 infection and that the latter might keep antiviral and inflammatory macrophage/DC functions silent. Although the slow replication kinetics of FL13 likely contribute to differences in cellular gene expression, the data suggest distinct mechanisms of interaction of the two viruses with the innate immune system during early infection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Crisci, Moroldo, Vu Manh, Mohammad, Jourdren, Urien, Bouguyon, Bordet, Bevilacqua, Bourge, Pezant, Pléau, Boulesteix, Schwartz, Bertho and Giuffra.)

DNA vaccination is an attractive technology, based on its well-established manufacturing process, safety profile, adaptability to rapidly combat pandemic pathogens, and stability at ambient temperature; however an optimal delivery method of DNA remains to be determined. As pigs are a relevant model for humans, we comparatively evaluated the efficiency of vaccine DNA delivery in vivo to pigs using dissolvable microneedle patches, intradermal inoculation with needle (ID), surface electroporation (EP), with DNA associated or not to cationic poly-lactic-co-glycolic acid nanoparticles (NPs). We used a luciferase encoding plasmid (pLuc) as a reporter and vaccine plasmids encoding antigens from the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), a clinically-significant swine arterivirus. Patches were successful at inducing luciferase expression in skin although at lower level than EP. EP induced the cutaneaous recruitment of granulocytes, of MHC2 pos CD172A pos myeloid cells and type 1 conventional dendritic cells, in association with local production of IL-1β, IL-8 and IL-17; these local responses were more limited with ID and undetectable with patches. The addition of NP to EP especially promoted the recruitment of the MHC2 pos CD172A pos CD163 int and CD163 neg myeloid subsets. Notably we obtained the strongest and broadest IFNγ T-cell response against a panel of PRRSV antigens with DNA + NPs delivered by EP, whereas patches and ID were ineffective. The anti-PRRSV IgG responses were the highest with EP administration independently of NPs, mild with ID, and undetectable with patches. These results contrast with the immunogenicity and efficacy previously induced in mice with patches. This study concludes that successful DNA vaccine administration in skin can be achieved in pigs with electroporation and patches, but only the former induces local inflammation, humoral and cellular immunity, with the highest potency when NPs were used. This finding shows the importance of evaluating the delivery and immunogenicity of DNA vaccines beyond the mouse model in a preclinical model relevant to human such as pig and reveals that EP with DNA combined to NP induces strong immunogenicity., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) induces reproductive disorders in sows and respiratory illnesses in growing pigs and is considered as one of the main pathogenic agents responsible for economic losses in the porcine industry worldwide. Modified live PRRSV vaccines (MLVs) are very effective vaccine types against homologous strains but they present only partial protection against heterologous viral variants. With the goal to induce broad and cross-protective immunity, we generated DNA vaccines encoding B and T antigens derived from a European subtype 1 strain that include T-cell epitope sequences known to be conserved across strains. These antigens were expressed either in a native form or in the form of vaccibodies targeted to the endocytic receptor XCR1 and CD11c expressed by different types of antigen-presenting cells (APCs). When delivered in skin with cationic nanoparticles and surface electroporation, multiple DNA vaccinations as a stand-alone regimen induced substantial antibody and T-cell responses, which were not promoted by targeting antigens to APCs. Interestingly, a DNA-MLV prime-boost strategy strongly enhanced the antibody response and broadened the T-cell responses over the one induced by MLV or DNA-only. The anti-nucleoprotein antibody response induced by the DNA-MLV prime-boost was clearly promoted by targeting the antigen to CD11c and XCR1, indicating a benefit of APC-targeting on the B-cell response. In conclusion, a DNA-MLV prime-boost strategy, by enhancing the potency and breadth of MLV vaccines, stands as a promising vaccine strategy to improve the control of PRRSV in infected herds.

Swine lymph nodes (LN) present an inverted structure compared to mouse and human, with the afferent lymph diffusing from the center to the periphery. This structure, also observed in close and distant species such as dolphins, hippopotamus, rhinoceros, and elephants, is poorly described, nor are the LN macrophage populations and their relationship with B cell follicles. B cell maturation occurs mainly in LN B cell follicles with the help of LN macrophage populations endowed with different antigen delivery capacities. We identified three macrophage populations that we localized in the inverted LN spatial organization. This allowed us to ascribe porcine LN MΦ to their murine counterparts: subcapsular sinus MΦ, medullary cord MΦ and medullary sinus MΦ. We identified the different intra and extrafollicular stages of LN B cells maturation and explored the interaction of MΦ, drained antigen and follicular B cells. The porcine reproductive and respiratory syndrome virus (PRRSV) is a major porcine pathogen that infects tissue macrophages (MΦ). PRRSV is persistent in the secondary lymphoid tissues and induces a delay in neutralizing antibodies appearance. We observed PRRSV interaction with two LN MΦ populations, of which one interacts closely with centroblasts. We observed BCL6 up-regulation in centroblast upon PRRSV infection, leading to new hypothesis on PRRSV inhibition of B cell maturation. This seminal study of porcine LN will permit fruitful comparison with murine and human LN for a better understanding of normal and inverted LN development and functioning.

Porcine Reproductive and Respiratory Syndrome virus (PRRSV) is an arterivirus responsible for highly contagious infection and huge economic losses in pig industry. Two species, PRRSV-1 and PRRSV-2 are distinguished, PRRSV-1 being more prevalent in Europe. PRRSV-1 can further be divided in subtypes. PRRSV-1.3 such as Lena are more pathogenic than PRRSV-1.1 such as Lelystad or Flanders13. PRRSV-1.3 viruses trigger a higher Th1 response than PRRSV-1.1, although the role of the cellular immune response in PRRSV clearance remains ill defined. The pathogenicity as well as the T cell response inductions may be differentially impacted according to the capacity of the virus strain to infect and/or activate DCs. However, the interactions of PRRSV with in vivo -differentiated-DC subtypes such as conventional DC1 (cDC1), cDC2, and monocyte-derived DCs (moDC) have not been thoroughly investigated. Here, DC subpopulations from Lena in vivo infected pigs were analyzed for viral genome detection. This experiment demonstrates that cDC1, cDC2, and moDC are not infected in vivo by Lena. Analysis of DC cytokines production revealed that cDC1 are clearly activated in vivo by Lena. In vitro comparison of 3 Europeans strains revealed no infection of the cDC1 and cDC2 and no or little infection of moDC with Lena, whereas the two PRRSV-1.1 strains infect none of the 3 DC subtypes. In vitro investigation of T helper polarization and cytokines production demonstrate that Lena induces a higher Th1 polarization and IFNγ secretion than FL13 and LV. Altogether, this work suggests an activation of cDC1 by Lena associated with a Th1 immune response polarization.

Lung inflammation is frequently involved in respiratory conditions and it is strongly controlled by mononuclear phagocytes (MNP). We previously studied porcine lung MNP and described a new population of cells presenting all the features of alveolar macrophages (AM) except for their parenchymal location, that we named AM-like cells. Herein we showed that AM-like cells are macrophages phagocytosing blood-borne particles, in agreement with a pulmonary intravascular macrophages (PIM) identity. PIM have been described microscopically long time ago in species from the Laurasiatheria superorder such as bovine, swine, cats or cetaceans. We observed that PIM were more inflammatory than AM upon infection with the porcine reproductive and respiratory syndrome virus (PRRSV), a major swine pathogen. Moreover, whereas PRRSV was thought to mainly target AM, we observed that PIM were a major producer of virus. The PIM infection was more correlated with viremia in vivo than AM infection. Finally like AM, PIM-expressed genes were characteristic of an embryonic monocyte-derived macrophage population, whose turnover is independent of bone marrow-derived hematopoietic precursors. This last observation raised the interesting possibility that AM and PIM originate from the same lung precursor.

In human medicine, bronchoalveolar lavage is the main non-traumatic procedure allowing an insight into the respiratory Dendritic Cells (DC) and Macrophages populations. However, it has never been demonstrated in a relevant model that alveolar DC subpopulations were comparable to their interstitial counterparts. In a precedent work we observed that respiratory pig DC and Macrophages were more similar to the human ones than to the mouse ones. In the present work, thanks to our animal model, we were able to collect the rare bronchoalveolar DC and compare them to their interstitial counterparts. We observed that DC presented very similar gene-expression patterns in the alveolar and interstitial compartments, validating the study of human bronchoalveolar DC as surrogate of their interstitium counterparts., Competing Interests: The authors have declared that no competing interests exist.

Previous studies have demonstrated that angiotensin-II (A-II) increases the human adrenocorticotropin receptor (hMC2R) gene expression in adrenal cells. In the present study, we have characterized two activator protein-1 (AP-1)-binding sites involved in the A-II stimulation of hMC2R gene transcription. Vectors containing different fragments of the hMC2R gene promoter inserted upstream of the luciferase gene, have been constructed. After transfection of H295R cells with these constructs and treatment of the cells by A-II during 48 h, maximal stimulation of the luciferase activity was obtained using the construct p(-263/+22)luc. Using progressively deleted constructs, three regions responsible for the A-II-stimulated transcription of hMC2R have been delineated. Inside these regions, two sequences displayed some homology with an AP-1 binding element (AP-108 and AP-203). Mutation of either AP-108 or AP-203 site induced a decrease of A-II-stimulated luciferase activity by 40% and 25%, respectively. Gel-shift analysis showed protein binding to these sites which was increased by an A-II treatment (maximum obtained after 3 h). Moreover, A-II could rapidly increase mRNA levels of several factors belonging to the Fos and Jun families which may be components of the AP-1 complex.