A loss of function mutation in SOCS2 results in increased inflammatory response of macrophages to TLR ligands and Staphylococcus aureus

IntroductionThe role of suppressor of cytokine signaling (SOCS)2 in anti-infective bacterial immunity has been poorly investigated compared to other members of the SOCS family.MethodsWe characterized the previously identified loss of function R96C point mutation of SOCS2 using a genome-edited mouse model that resumes the phenotype of Socs2 knockout mice. The response of macrophages to TLR-ligands and Staphylococcus aureus was examined.Results and discussionConversely to previously published data using human monocyte-derived macrophages, the stimulation of bone-marrow-derived macrophages with various TLR ligands did not show any difference according to the SOCS2 variant. Upregulation of IL-6 and TNF-α pro-inflammatory cytokines production was only seen when the SOCS2 expression was promoted by the culture of macrophages in the presence of GM-CSF. Furthermore, we showed that the SOCS2 point mutation is associated with heightened STAT5 phosphorylation in a short time frame upon GM-CSF incubation. In mice, recruitment of neutrophil and F4/80int Ly6C+ inflammatory macrophage, as well as IFN-γ and IL-10 concentrations, are significantly increased upon S. aureus peritoneal infection. Altogether, these data support the idea that by lowering the pro-inflammatory environment, SOCS2 favors better control of bacterial burden during a systemic infection caused by S. aureus

Epidémiologie moléculaire et évolution de l'entérovirus A71 et interactions génétiques avec les autres entérovirus de l'espèce A responsables de la maladie pied-main-bouche.

Hand-Foot and Mouth Disease (HFMD) and Herpangina are two benign pediatric diseases caused by Enteroviruses (EV), especially enterovirus A species serotypes (EV-A). Infections caused by the EV-A71 serotype are monitored in countries of South East Asia because they are associated with severe neurological symptoms in young children and may be fatal (cardiopulmonary failure). Infections caused by the other EV-A serotypes, e.g. coxsackievirus A16 (CV-A16), rarely induce severe symptoms. In Europe, EV-A71 HFMD cases are not notifiable because this virus does not cause large-scale epidemics. The overall objective of this thesis was to study the EV-A epidemiology in Europe and we used a Bayesian phylogenetic approach to analyze 500 viral strains. We show a discontinued circulation of two EV-A71 populations (C1 and C2 subgenogroups), which explains the rare outbreaks in Europe. The epidemiology of this virus is characterized by transportation events of viral strains between European countries and sporadically between Europe and Asia (C4 and B5 subgenogroups). Intertypic genetic recombination occur rarely among circulating EV-A71 populations and does not contribute significantly to their genetic diversity. We found that genetic mechanism was related to the emergence of a new CV-A16 subgenogroup, which is circulating in France since 2011. In comparison with EV-A71, a number of serotypes (CV-A2, CV-A4, and CV-A6) are more frequently involved in intertypic recombination events. The structural and functional constraints are possible factors involved in the slow mutation fixation in the EV-A71 capsid proteins as determined by analyses of molecular selection. Neurovirulence, the recent and repeated introductions of variants “Asian” strains, and the diversity of genogroups in Africa and India call for strengthened surveillance of EV-A71 infections among European countries.La maladie pied-main-bouche (PMB) et l’herpangine sont deux maladies pédiatriques bénignes causées par les entérovirus (EV), en particulier les sérotypes de l’espèce A (EV-A). Le sérotype EV-A71 fait l’objet d’une surveillance dans les pays du Sud Est de l’Asie car il est associé à des atteintes neurologiques sévères chez les très jeunes enfants, parfois mortelles (défaillance cardio-pulmonaire). Les infections causées par les autres EV-A tel que le coxsackievirus A16 (CV-A16) provoquent rarement des atteintes sévères. En Europe, les cas de maladie PMB causés par l’EV-A71 ne font pas l’objet d’une déclaration obligatoire, car ce virus ne cause pas d’épidémies de grande ampleur. L’objectif général de la thèse était d’étudier l’épidémiologie des EV-A en Europe et nous avons utilisé une approche phylogénétique bayésienne pour analyser un échantillon de 500 souches. Nous montrons la circulation discontinue de l’EV-A71 de deux populations virales principales (sous génogroupes C1 et C2), ce qui explique la rareté des épidémies en Europe. L’épidémiologie de ce virus est aussi caractérisée par des transports de souches entre les pays Européens et sporadiquement entre l’Europe et l’Asie (sous génogroupes B5 et C4). La recombinaison génétique intertypique survient rarement parmi les populations d’EV-A71 en circulation et ne contribue pas significativement à leur diversité génétique. Cependant, ce mécanisme génétique est relié à l’émergence d’un sous génogroupe CV-A16 qui circule en France depuis 2011. Comparés à l’EV-A71, les sérotypes CV-A2, CV-A4, CV-A6 sont plus fréquemment sujets à des événements de recombinaison intertypiques. L’analyse de la sélection à l’échelle moléculaire indique que la fixation des mutations dans les protéines de capside de l’EV-A71 est lente, probablement à cause des contraintes structurales et fonctionnelles. La surveillance des infections à EV-A71 en Europe devrait être renforcée à cause de la neurovirulence de ce virus, de l’introduction récente et répétée de souches variantes « asiatiques » et de l’existence d’une grande diversité de génogroupes en Afrique et en Inde encore peu explorée

Molecular epidemiology and evolution of enterovirus A71 and genetic interactions with others enterovirus A species responsive of Hand-Foot and Mouth Disease

La maladie pied-main-bouche (PMB) et l’herpangine sont deux maladies pédiatriques bénignes causées par les entérovirus (EV), en particulier les sérotypes de l’espèce A (EV-A). Le sérotype EV-A71 fait l’objet d’une surveillance dans les pays du Sud Est de l’Asie car il est associé à des atteintes neurologiques sévères chez les très jeunes enfants, parfois mortelles (défaillance cardio-pulmonaire). Les infections causées par les autres EV-A tel que le coxsackievirus A16 (CV-A16) provoquent rarement des atteintes sévères. En Europe, les cas de maladie PMB causés par l’EV-A71 ne font pas l’objet d’une déclaration obligatoire, car ce virus ne cause pas d’épidémies de grande ampleur. L’objectif général de la thèse était d’étudier l’épidémiologie des EV-A en Europe et nous avons utilisé une approche phylogénétique bayésienne pour analyser un échantillon de 500 souches. Nous montrons la circulation discontinue de l’EV-A71 de deux populations virales principales (sous génogroupes C1 et C2), ce qui explique la rareté des épidémies en Europe. L’épidémiologie de ce virus est aussi caractérisée par des transports de souches entre les pays Européens et sporadiquement entre l’Europe et l’Asie (sous génogroupes B5 et C4). La recombinaison génétique intertypique survient rarement parmi les populations d’EV-A71 en circulation et ne contribue pas significativement à leur diversité génétique. Cependant, ce mécanisme génétique est relié à l’émergence d’un sous génogroupe CV-A16 qui circule en France depuis 2011. Comparés à l’EV-A71, les sérotypes CV-A2, CV-A4, CV-A6 sont plus fréquemment sujets à des événements de recombinaison intertypiques. L’analyse de la sélection à l’échelle moléculaire indique que la fixation des mutations dans les protéines de capside de l’EV-A71 est lente, probablement à cause des contraintes structurales et fonctionnelles. La surveillance des infections à EV-A71 en Europe devrait être renforcée à cause de la neurovirulence de ce virus, de l’introduction récente et répétée de souches variantes « asiatiques » et de l’existence d’une grande diversité de génogroupes en Afrique et en Inde encore peu explorée.Hand-Foot and Mouth Disease (HFMD) and Herpangina are two benign pediatric diseases caused by Enteroviruses (EV), especially enterovirus A species serotypes (EV-A). Infections caused by the EV-A71 serotype are monitored in countries of South East Asia because they are associated with severe neurological symptoms in young children and may be fatal (cardiopulmonary failure). Infections caused by the other EV-A serotypes, e.g. coxsackievirus A16 (CV-A16), rarely induce severe symptoms. In Europe, EV-A71 HFMD cases are not notifiable because this virus does not cause large-scale epidemics. The overall objective of this thesis was to study the EV-A epidemiology in Europe and we used a Bayesian phylogenetic approach to analyze 500 viral strains. We show a discontinued circulation of two EV-A71 populations (C1 and C2 subgenogroups), which explains the rare outbreaks in Europe. The epidemiology of this virus is characterized by transportation events of viral strains between European countries and sporadically between Europe and Asia (C4 and B5 subgenogroups). Intertypic genetic recombination occur rarely among circulating EV-A71 populations and does not contribute significantly to their genetic diversity. We found that genetic mechanism was related to the emergence of a new CV-A16 subgenogroup, which is circulating in France since 2011. In comparison with EV-A71, a number of serotypes (CV-A2, CV-A4, and CV-A6) are more frequently involved in intertypic recombination events. The structural and functional constraints are possible factors involved in the slow mutation fixation in the EV-A71 capsid proteins as determined by analyses of molecular selection. Neurovirulence, the recent and repeated introductions of variants “Asian” strains, and the diversity of genogroups in Africa and India call for strengthened surveillance of EV-A71 infections among European countries

Thymus-specific serine protease, a protease that shapes the CD4 T cell repertoire

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Dynamique des microbiomes des voies respiratoires supérieures et inférieures associée à l'asthme sévère du nourrisson

International audienceThe human respiratory tract is colonized by microorganisms which constitute the respiratory microbiota. This microbiota is involved in the protection of the host against microbial infections. Its dynamics is regulated by mechanisms such as respiratory air flows, contributing to the inhalation of microorganisms, which is balanced by the host's immune system. An imbalance of these microbial communities, called dysbiosis, seems to be involved in the occurrence of respiratory pathologies such as asthma. This pathology affects about 25% of infants and its pathophysiology suggests a microbiota damage, associated with an inflammation. Major aims of this project are to characterize the global composition of the respiratory microbiome of infants with severe asthma, to follow its evolution over time and to determine the inflammatory profile of these patients. The cohort will include 100 patients from University Hospitals of Rouen and Caen. Nasopharyngeal and bronchoalveolar samples from these patients will be sequenced by metatranscriptomic approach and will allow to describe the respiratory microbiome composition. Inflammatory markers will also be analyzed through flow cytometry and ELISA to identify links between inflammation profiles involved in severe asthma and microbiome composition

"Microbiosthme" : Upper and lower airway microbiomes analysis in infants with severe asthma

International audienceThe human respiratory tract is colonized by microorganisms which constitute the respiratory microbiota. This microbiota is involved in the protection of the host against microbial infections. Its dynamics is regulated by mechanisms such as respiratory air flows, contributing to the inhalation of microorganisms, which is balanced by the host's immune system. An imbalance of these microbial communities, called dysbiosis, seems to be involved in the occurrence of respiratory pathologies such as asthma. This pathology affects about 25% of infants and its pathophysiology suggests a microbiota damage, associated with an inflammation. Major aims of this project are to characterize the global composition of the respiratory microbiome of infants with severe asthma, to follow its evolution over time and to determine the inflammatory profile of these patients. The cohort will include 100 patients from University Hospitals of Rouen and Caen. Nasopharyngeal and bronchoalveolar samples from these patients will be sequenced by metatranscriptomic approach and will allow to describe the respiratory microbiome composition. Inflammatory markers will also be analyzed through flow cytometry and ELISA to identify links between inflammation profiles involved in severe asthma and microbiome composition

Tissue-Specific Factors Differentially Regulate the Expression of Antigen-Processing Enzymes During Dendritic Cell Ontogeny

International audienceDendritic cells (DCs) form a collection of antigen-presenting cells (APCs) that are distributed throughout the body. Conventional DCs (cDCs), which include the cDC1 and cDC2 subsets, and plasmacytoid DCs (pDCs) constitute the two major ontogenically distinct DC populations. The pDCs complete their differentiation in the bone marrow (BM), whereas the cDC subsets derive from pre-committed BM precursors, the pre-cDC, that seed lymphoid and non-lymphoid tissues where they further differentiate into mature cDC1 and cDC2. Within different tissues, cDCs express distinct phenotype and function. Notably, cDCs in the thymus are exquisitely efficient at processing and presenting antigens in the class II pathway, whereas in the spleen they do so only upon maturation induced by danger signals. To appraise this functional heterogeneity, we examined the regulation of the expression of distinct antigen-processing enzymes during DC ontogeny. We analyzed the expression of cathepsin S (CTSS), cathepsin L (CTSL), and thymus-specific serine protease (TSSP), three major antigen-processing enzymes regulating class II presentation in cDC, by DC BM precursors and immature and mature cDCs from the spleen and thymus. We found that pre-cDCs in the BM express relatively high levels of these different proteases. Then, their expression is modulated in a tissue-specific and subset-specific manner with immature and mature thymic cDCs expressing overall higher levels than immature splenic cDCs. On the other hand, the TSSP expression level is selectively down-regulated in spleen pDCs, whereas CTSS and CTSL are both increased in thymic and splenic pDCs. Hence, tissue-specific factors program the expression levels of these different proteases during DC differentiation, thus conferring tissue-specific function to the different DC subsets

Etude préliminaire du microbiote urinaire bactérien de femmes non ménopausées

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Citrulline protects human retinal pigment epithelium from hydrogen peroxide and iron/ascorbate induced damages

International audienceAbstract Oxidative stress plays an important role in the ageing of the retina and in the pathogenesis of retinal diseases such as age‐related macular degeneration (ARMD). Hydrogen peroxide is a reactive oxygen species generated by the photo‐excited lipofuscin that accumulates during ageing in the retinal pigment epithelium (RPE), and the age‐related accumulation of lipofuscin is associated with ARMD. Iron also accumulates with age in the RPE that may contribute to ARMD as an important source of oxidative stress. The aim of this work was to investigate the effects of L‐Citrulline (CIT), a naturally occurring amino acid with known antioxidant properties, on oxidative stressed cultured RPE cells. Human RPE (ARPE‐19) cells were exposed to hydrogen peroxide (H 2 O 2 ) or iron/ascorbate (I/A) for 4 h, either in the presence of CIT or after 24 h of pretreatment. Here, we show that supplementation with CIT protects ARPE‐19 cells against H 2 O 2 and I/A. CIT improves cell metabolic activity, decreases ROS production, limits lipid peroxidation, reduces cell death and attenuates IL‐8 secretion. Our study evidences that CIT is able to protect human RPE cells from oxidative damage and suggests potential protective effect for the treatment of retinal diseases associated with oxidative stress