Revue des maladies respiratoires最新文献

{"title":"Characterization an in vitro 3D alveolar model in COPD","authors":"A. Bodin ,&nbsp;A. Lan ,&nbsp;P. Bellaud ,&nbsp;V. Lagente ,&nbsp;T. Victoni","doi":"10.1016/j.rmr.2024.01.021","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.021","url":null,"abstract":"<div><h3>Introduction</h3><p>New interest emerges on organoids for studying lung diseases, mainly chronic obstructive pulmonary disease (COPD). COPD is lung disease a characterize by chronic inflammation and excessive remodeling in small airway. The complexity of this disease is now widely recognized, cellular-based research remains highly challenging because of the lack of suitable experimental models that recapitulate between interaction of cells. Alveolar organoids could enable us to assess the involvement of several type cells in response to damage from cigarette smoke, one major risk factor of COPD. The aim of this study is to characterize an in vitro 3D alveolar model from mouse lungs.</p></div><div><h3>Methods</h3><p>We used progenitor cells isolated by filtration from pieces of mouse lung and cultivated in Matrigel. We cultivated first part of lung organoids in conditioned expansion medium for ten days. After another part of organoids were differentiated in conditioned differentiation alveolar medium during ten days. Then, we assessed several markers of alveolar cells differentiated, fibroblast markers, or stem cell markers gene expression by RT-qPCR in the two conditions.</p></div><div><h3>Results</h3><p>The markers used are as follow: (1) aquaporin 5 (Aqp5), homeodomain-only protein homeobox (Hopx), advanced glycation endproduct-specific receptor (Ager) and podoplanin (Pdpn) for alveolar type 1 (AT1); (2) surfactant protein b (Sfptb), surfactant protein c (Sfptc) for alveolar type 2 (AT2); (3) platelet-derived growth factor alpha (Pdgfrα) for fibroblast; (4) secretoglobin family 1A member 1 (Scgb1a1) and surfactant protein c (Sfptc) for stem cells; (5) epithelial cell adhesion molecule (Epcam) as markers of differentiated airway and alveolar epithelium. Epcam, Sfptb and Aqp5 gene expression was increased in both conditions compared to the single cell group. On the other hand, aquaporin 5 expression was greater in the differentiated medium condition compared with organoids exposed to undifferentiated medium. We also observed an increase in the expression of the Sfptc marker in the differentiated condition. Futhermore, fibroblast and stem cells appear to be poorly represented in organoids, indeed the following markers platelet-derived growth factor alpha (Pdgfrα) and secretoglobin family 1A member 1 (Scgb1a1) are weakly expressed.</p></div><div><h3>Conclusion</h3><p>Our results show alveolar organoids derived mouse lung present both AT1 and AT2 cells. So, these organoids can used as multicellular experimental models for studying cigarette smoke damage.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 191"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sputum as a new reflective tool of local immune response in lung cancer","authors":"M. Ferreira ,&nbsp;M. Pronost ,&nbsp;L. Guillot ,&nbsp;D. Sizaret ,&nbsp;A. Gey ,&nbsp;E. Tartour ,&nbsp;T. Secher ,&nbsp;N. Heuzé-Vourc’h","doi":"10.1016/j.rmr.2024.01.032","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.032","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;The management of metastatic non-small cell lung cancer (NSCLC) has been revolutionized by immune checkpoint inhibitors (ICI). However, it would be valuable to refine ICI treatment with predictive combined biomarkers models. The tumor microenvironment is profoundly affecting ICI response in NSCLC. Tissue-resident memory T cells (TRM) are a non-circulating lymphocyte population encountered in peripheral tissues with both memory and effector functions. TRM infiltration in lung tumor is correlated with favourable ICI response in NSCLC. But it is not possible to perform iterative lung biopsies to monitor TRM during treatment. The objectives of this study were to evaluate the feasibility of a longitudinal collection of sputum in NSCLC patients and to evaluate TRM cells in comparison with lung biopsies.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;p&gt;This prospective and exploratory study included 9 patients with a NSCLC treated in first line with pembrolizumab. Before the first ICI infusion and every treatment day until 12-month follow-up, patients received a 15-minute aerosol of isotonic saline solution during which they expectorate. To recover sputum cells, samples were clarified from mucus, stirred, and filtered. TRM cells were quantified by flow cytometry, as CD&lt;sub&gt;45&lt;/sub&gt; +CD&lt;sub&gt;3&lt;/sub&gt; +CD&lt;sub&gt;8&lt;/sub&gt; +CD&lt;sub&gt;49&lt;/sub&gt; +CD&lt;sub&gt;69&lt;/sub&gt; +CD&lt;sub&gt;103&lt;/sub&gt;+ or CD&lt;sub&gt;103&lt;/sub&gt;− cells. Spectral multiplex in situ immunofluorescence was used to quantify TRM cells in lung tissue biopsies collected before ICI treatment.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;p&gt;Between May 2021 and September 2022. Nine patients were included: median age was 62 (42–83) years and all patients had smoking history. All patients were able to provide a sputum at each visit and the median follow-up duration was 6.9&lt;!--&gt; &lt;!--&gt;months. We were able to identify longitudinally TRM cells in sputum: 80 adequate sputum samples (97.6%) were analyzed by flow cytometry. In order to evaluate the representativeness of TRM cells in sputum, we compared TRM cell proportion determined in lung biopsies by immunofluorescence (method 1) and in the first sputum, by flow cytometry (method 2). The Bland-Altman analysis showed that differences between the 2 methods fitted in the 95% of agreement and bias were close to zero. Thus, there was an agreement between the measurement methods. Moreover, calculation of Pearson coefficient showed a &lt;em&gt;P&lt;/em&gt;-value&lt;!--&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0.05 for both CD&lt;sub&gt;103&lt;/sub&gt;+ and CD&lt;sub&gt;103&lt;/sub&gt;− cells, suggesting that the correlation between the two methods was significant.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;p&gt;Our results indicate that sputum offer a simple and non-invasive indirect estimate of the immune cell infiltration in the pulmonary tissue and allow analysis of TRM cell dynamics during ICI treatment in NSCLC. Sputum collection might help understanding evolution of local immune response during NSCLC and ICI treatment, extending immunological studies to","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 196-197"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rôle de la dysfonction du facteur d’angiogenèse VEGF dans la sénescence des cellules endothéliales pulmonaires","authors":"J. Jacquet ,&nbsp;E. Marcos ,&nbsp;L. Lipskaia ,&nbsp;V. Gros ,&nbsp;E. Born ,&nbsp;N. Vienney ,&nbsp;A. Houssaini ,&nbsp;C. Jourdan Le Saux ,&nbsp;S. Adnot ,&nbsp;L. Boyer","doi":"10.1016/j.rmr.2024.01.035","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.035","url":null,"abstract":"<div><h3>Introduction</h3><p>Le vieillissement vasculaire entraîne une raréfaction des microvaisseaux. Le mécanisme proposé est une insuffisance de signalisation du facteur de croissance endothélial vasculaire (VEGF) dont l’action est perturbée par une production accrue d’une forme soluble de son récepteur VEGFR1 (sVEGFR1) qui l’empêche de se fixer à son récepteur actif, le VEGFR1 exprimé par les CEs. Les CEs des microvaisseaux pulmonaires (PCEs) sont nécessaires à la fonction des poumons et nécessitent le signal de survie du VEGF, en l’absence duquel elles peuvent rentrer en sénescence, caractérisée par un arrêt de la division cellulaire et la libération de facteurs nocifs pour les poumons. Nous avons montré une accumulation de PCEs sénescentes au cours de l’hypertension (HTP) et de l’emphysème pulmonaires.</p></div><div><h3>Méthode</h3><p>Évaluer l’insuffisance de signalisation du VEGF dans la sénescence des PCEs liée à deux pathologies, l’hypertension (HTP) et l’emphysème pulmonaires.</p></div><div><h3>Résultats</h3><p>Les poumons des souris âgées par rapport aux jeunes souris ont une densité capillaire plus faible se manifestant par une diminution du nombre de PCEs identifiées en immunohistochimie par marquage de l’ICAM1 ou de l’isolectine B4. Le traitement des souris par l’inhibiteur du récepteur du VEGF Sugen amplifie ces effets et augmente le nombre de PCEs sénescentes marquées p16, en diminuant le réseau capillaire pulmonaire, en détériorant l’hémodynamique pulmonaire et en induisant de l’emphysème. L’expression pulmonaire de sVEGFR1 chez les souris développant une HTP hypoxique est augmentée et aggravée par le Sugen, de même que l’expression pulmonaire de deux facteurs modulant l’épissage alternatif du VEGFR1 et la formation de sVEGFR1, le domaine Jumonji contenant la protéine 6 (Jmjd6) et le TNFSF15. Nous montrons in vitro qu’un traitement par le sVEGFR1 de PCEs en culture issues de patients (étude Costemcells) entraîne une augmentation de la sénescence cellulaire. Pour contrer la forte susceptibilité des PCEs à la sénescence dans l’HTP et l’emphysème, nous avons généré un nouveau modèle murin qui surexprime le VEGF selon un système Tet On inductible et spécifique au tissu. Ces souris sont en cours d’étude, croisées avec des souris exprimant la Cre-recombinase dans le foie (souris Alb-Cre), afin d’assurer un apport contrôlé de VEGF du foie vers les poumons.</p></div><div><h3>Conclusion</h3><p>L’augmentation du sVEGFR1 circulant au cours du vieillissement et des pathologies pulmonaires tels que l’HTP et l’emphysème pourrait être responsable de la sénescence des PCEs. Contrecarrer l’insuffisance de signalisation du VEGF devrait permettre de réduire la sévérité de ces pathologies.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 198"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Implication des protéines JAK2 et STAT-1/3/5 dans la sarcoïdose","authors":"T. Iskounen ,&nbsp;R. Hindre ,&nbsp;F. Jeny ,&nbsp;O. Frenoy ,&nbsp;M.N. Triba ,&nbsp;J.F. Bernaudin ,&nbsp;D. Valeyre ,&nbsp;M. Kambouchner ,&nbsp;H. Nunes ,&nbsp;C. Planès ,&nbsp;V. Besnard","doi":"10.1016/j.rmr.2024.01.069","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.069","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;La sarcoïdose est une maladie systémique d’étiologie inconnue caractérisée par la formation de granulomes immuns, principalement dans le poumon. L’analyse génomique du sang périphérique a montré que la voie de signalisation JAK/STAT est significativement représentée dans la sarcoïdose. Notre objectif était de déterminer l’expression des membres de la voie JAK/STAT au cours du développement des granulomes chez des patients présentant différents stades de sévérité de la pathologie.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Méthodes&lt;/h3&gt;&lt;p&gt;Une étude rétrospective a été réalisée sur des biopsies de patients diagnostiqués pour la sarcoïdose. Les formes activées des protéines JAKs et STATs ont été étudiées par immunodétection (histochimie, fluorescence, Hyperion) sur des biopies : poumon (alvéolaire) (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;10), peau (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;13), ganglions lymphatiques (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;12), foie (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;9) et glandes salivaires (n&lt;!--&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;8). Des tests statistiques ont été réalisés pour comparer l’activation des protéines JAKs et STATs dans les granulomes de différents organes, et étudier leurs relations avec le stade évolutif du granulome (naissant/floride/« naked »/fibreux).&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Résultats&lt;/h3&gt;&lt;p&gt;La cohorte regroupait deux profils d’évolution variable : sarcoïdose résolue (50 %) ou persistante (50 %). Seule pJAK2 a été détectée, principalement dans les lymphocytes CD3+ et peu dans la lignée myéloïde. Dans les lymphocytes, seule une localisation nucléaire pJAK2 a été observée, suggérant l’activation d’une voie non canonique de JAK2. La localisation nucléaire de pJAK2 a été confirmée par colocalisation avec DAPI/histone H3. pJAK2 est principalement colocalisée avec l’histone H3K4me, associée à une activité transcriptionnelle intense. Seules les formes actives de pSTAT1, pSTAT3 et pSTAT5 ont été détectées dans les granulomes, principalement dans les cellules de la lignée myéloïde. Les formes actives pJAK2 et pSTAT1/pSTAT3/pSTAT5 ont des niveaux d’expression variables dans les biopsies analysées. PJAK2 est exprimée quasi-systématiquement dans toutes les biopsies, alors que pSTAT1, pSTAT3 et pSTAT5 sont exprimés au moins dans la moitié des biopsies. L’expression des 4 protéines est la plus importante dans les biopsies pulmonaires (AUC&lt;!--&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0,83) et cutanées (AUC&lt;!--&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0,89) et est en moyenne associée à une persistance de la maladie (COS&lt;!--&gt; &lt;!--&gt;≥&lt;!--&gt; &lt;!--&gt;5, &lt;em&gt;p&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0,001). Deux signatures ont été identifiées dans les lymphocytes/lignée myéloïde : pJAK2/pSTAT1-pSTAT5 (poumon ; (AUC&lt;!--&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0,84)) et pJAK2/pSTAT3-pSTAT5 (peau ; [AUC&lt;!--&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0,80]). L’analyse des stades évolutifs du granulome a montré une corrélation positive significative entre l’expression des formes activées de pSTAT1(&lt;em&gt;p&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0,05), pSTAT3 (&lt;em&gt;p&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0,05), pSTAT5 (&lt;em&gt;","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 215-216"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition de Gp96 comme stratégie anti-fibrotique dans la fibrose pulmonaire idiopathique et son suivi par l’imagerie SPECT in vivo de la Fibroblast Activation Protein","authors":"A. Dias ,&nbsp;H. Sikner ,&nbsp;A.G. Garnier ,&nbsp;J. Tanguy ,&nbsp;A. Bouchard ,&nbsp;M. Moreau ,&nbsp;M. Claron ,&nbsp;O. Burgy ,&nbsp;F. Goirand ,&nbsp;P. Bonniaud ,&nbsp;B. Collin ,&nbsp;P.S. Bellaye","doi":"10.1016/j.rmr.2024.01.071","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.071","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;La fibrose pulmonaire idiopathique (FPI) est une maladie évolutive caractérisée par une production et un dépôt excessif de matrice extracellulaire (MEC) entraînant une insuffisance respiratoire chez les patients. Sous l’influence de cytokines pro-fibrotiques telles que le TGF-β1, les cellules pulmonaires vont acquérir un phénotype myofibroblastique caractérisé par l’expression de la &lt;em&gt;Fibroblast Activation Protein&lt;/em&gt; (FAP) et une surproduction de MEC. Les options de traitement de la FPI sont limitées et ne peuvent que retarder la progression de la maladie sans l’arrêter. La Gp96 est une protéine chaperonne du réticulum endoplasmique surexprimée au cours de la fibrose et favorisant l’expression membranaire de nombreux récepteurs impliqués dans la fibrogenèse. L’inhibition de Gp96 apparait comme une stratégie thérapeutique intéressante dans la FPI et nous émettons l’hypothèse que l’imagerie &lt;em&gt;in vivo&lt;/em&gt; des myofibroblastes via une sonde ciblant FAP pourrait permettre de suivre l’efficacité de ce type de traitement.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Méthodes&lt;/h3&gt;&lt;p&gt;Des souris C57BL/6 (mâles, 8 semaines) ont reçu une administration unique intratrachéale de bléomycine (BLM, 1,5&lt;!--&gt; &lt;!--&gt;mg/kg) ou de NaCl (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;14) comme contrôle, puis ont été traitées ou non par un inhibiteur de Gp96 (PU-WS13, 12,5&lt;!--&gt; &lt;!--&gt;mg/kg, gavage quotidien, &lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;17) ou du NaCl (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;16, gavage quotidien) de J&lt;sub&gt;8&lt;/sub&gt; à J&lt;sub&gt;22&lt;/sub&gt;. La fibrose a été suivie longitudinalement par imagerie CT à J&lt;sub&gt;8&lt;/sub&gt;, J&lt;sub&gt;15&lt;/sub&gt;et J&lt;sub&gt;22&lt;/sub&gt;. À J&lt;sub&gt;22&lt;/sub&gt;, les poumons et les liquides de lavage bronchoalvéolaires (BAL) ont été récupérés pour quantification du collagène (Rouge Sirius, Sircol et Ashcroft score), immunomarquage de FAP (IHC) et dosage du TGF-β1 (ELISA). &lt;em&gt;In vitro&lt;/em&gt;, des cellules A549 stimulées au TGF-β1 (2&lt;!--&gt; &lt;!--&gt;ng/mL, 24&lt;!--&gt; &lt;!--&gt;h) ont été traitées ou non par du PUWS13 (25/50&lt;!--&gt; &lt;!--&gt;μM, 24&lt;!--&gt; &lt;!--&gt;h) pour étudier l’expression de la protéine FAP (WB) et les interactions protéine-protéine entre Gp96 et FAP (Proximity ligation assay, PLA). Une sonde d’imagerie spécifique de FAP a été mise au point à partir d’un fragment d’anticorps anti-FAP couplé à un agent chélateur (DOTAGA) pour le radiomarquage à l’indium-111 pour l’imagerie par tomographie par émission monophotonique (SPECT). La sonde &lt;sup&gt;111&lt;/sup&gt;In-DOTAGA-Fab-FAP a été injectée par voie intraveineuse à des souris C57BL/6 (100&lt;!--&gt; &lt;!--&gt;μL, 10MBq, 25&lt;!--&gt; &lt;!--&gt;μg). Les souris ont subi des imageries SPECT à 1&lt;!--&gt; &lt;!--&gt;h, 4&lt;!--&gt; &lt;!--&gt;h et 24&lt;!--&gt; &lt;!--&gt;h post-injection à J&lt;sub&gt;7&lt;/sub&gt;et J&lt;sub&gt;15&lt;/sub&gt;. Des régions d’intérêt 3D ont été tracées sur le volume pulmonaire total (Vivoquant) afin de quantifier la fibrose (image CT) et l’accumulation de la sonde au niveau pulmonaire (SPECT) dans les différents groupes de souris (1/NaCl, 2/BLM et 3/BLM&lt;!--&gt; &lt;!--&gt;+&lt;!--&gt; &lt;!--&gt;PU-WS13).&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Résult","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 216-217"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of immunological biomarkers of macrophages related to specific compartmentalization of in lung and liver in mouse model of septic shock","authors":"A. Bodin ,&nbsp;C. Slek ,&nbsp;M. Magnin ,&nbsp;L. Commin ,&nbsp;A. Corlu ,&nbsp;V. Lagente ,&nbsp;C. Aninat ,&nbsp;J.M. Bonnet ,&nbsp;B. Allaouchiche ,&nbsp;V. Louzier ,&nbsp;T. Victoni","doi":"10.1016/j.rmr.2024.01.054","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.054","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;Sepsis is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection. The complex pathophysiology of sepsis is associated with pro- and anti-inflammatory response, a pro-coagulant state, endothelial dysfunction and tissue hypoxia. These mechanisms lead to progressive multi organ failure. Although this is a systemic process, the pathophysiological of sepsis differs from organ to organ, and from organ to peripheral blood. Our hypothesis to explain this compartmentalization of responses is a distinct population of resident tissue macrophages, as well as a distinct migration of monocytes in target organ. Indeed, the macrophages and monocytes can start the clinical syndrome of sepsis via transcription of genes involved in inflammation. Moreover, macrophages can induce endothelial injury by release reactive oxygen species. Also hypoxia decreases expression of M1 polarization markers and increases the M2 marker. Identification of biomarkers related to specific organs (beyond the blood) will improve the understanding specific organ failure. In this way, the aim of this study was compared to the systemic inflammatory response with the lung and the liver, two organs most affected during sepsis as well as understanding the role of macrophages in this compartmentalization. For this, a murine polymicrobial sepsis model induced by caecal ligation and puncture (CLP) was used.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;p&gt;Moderate sepsis was induced by the CLP in C57BL/6 male mice (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;63), divided into 4 groups: Basal, Sham 1 day and 5 days, CLP 1 days and 5 days. Then, we analyzed histological changes, cytokine profile (by ELISA and PCR), oxidative imbalance (expression of SOD, CAT and iNOS) and polarization markers (CD&lt;sub&gt;86&lt;/sub&gt;and CD&lt;sub&gt;206&lt;/sub&gt;) by immunohistochemistry.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;p&gt;Respiratory and liver failure was confirmed by histology and also by a decrease of pressure oxygen in arterial blood (PaO&lt;sub&gt;2&lt;/sub&gt;) and increase of bilirubin level after 5 days of CLP. Moreover CLP induced a plasma increase in the level of TNFα, IL-6, IL-10, KC and CCL2 in the first 24&lt;!--&gt; &lt;!--&gt;hours after CLP, but with a progressive decrease at 5 days. On the other hand, our original results show that the level of some of the cytokines in the liver and the lung differ from the systemic level during sepsis. Indeed, there are increase in CCL2 and its receptor CCR2 in the lung compared to the liver, whereas in the liver rather a decrease in the expression of CX3CL1/CX3CR1, that is not altered in the lung. We were also able to highlight a more marked oxidative imbalance in the liver than in the lung. Also, we observed that CLP increases the expression of M1 CD&lt;sub&gt;86&lt;/sub&gt; and M2 CD&lt;sub&gt;206&lt;/sub&gt; markers in the liver and lung at 5 days, but with a total number of CD&lt;sub&gt;86&lt;/sub&gt;-labeled cells, three times greater in the liver.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;p&gt;These results","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 207-208"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Thorough assessment of primary cell cultures for murine and human alveolar macrophages","authors":"C. David ,&nbsp;D. Brea ,&nbsp;A. Cezard ,&nbsp;V. Vasseur ,&nbsp;E. Barsac ,&nbsp;B. Briard ,&nbsp;M. Ferreira ,&nbsp;S. Adam-Marchand ,&nbsp;M. Si-Tahar ,&nbsp;A. Guillon","doi":"10.1016/j.rmr.2024.01.060","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.060","url":null,"abstract":"<div><h3>Introduction</h3><p>Alveolar macrophages (AM) are the first-line lung defenders, but appropriate cell culture models are still limited for their study. The uses of freshly isolated macrophages are hampered by limited quantities or difficult access to human donors. Indeed, research on AM is often restricted to cells differentiated in vitro from bone-marrow progenitors or blood monocytes; however, such cells do not present the specific features of tissue-specialized macrophages, like AM. To overcome these issues, Max Planck Institute (MPI) cells were characterized as a nontransformed, self-renewing AM-like macrophage murine model. Yet, MPI cannot fully reproduce AM characteristics because they developed outside the alveolar niche. To optimize the primary cell cultures of mouse AM (mAM), recent works have proposed strategies to expand and maintain these cells <em>ex vivo</em> from bronchoalveolar lavage (BAL). The functional impact of the amplification process is unclear. Moreover, the heterogeneity of AM population, which is constantly remodeled during the lifespan, is poorly recapitulated in laboratory mice, therefore limiting extrapolation of mAM physiopathology to human. Here, we propose a comprehensive comparison of mice and human AM (hAM) culture models and give insight into the translational value of different mice AM models.</p></div><div><h3>Methods</h3><p><em>Ex vivo</em> primary mAM were isolated from BAL of 3–5 mice and expanded for 3–4 weeks per passage. hAM were freshly isolated and maintained from BAL of uninfected patients. Immunophenotypes of MPI, mAM and hAM were compared. We evaluated the cells production of inflammatory mediators (IL-6, TNF-α, IL-1β) in response to different pro-inflammatory agonists such as LPS, poly (I: C), flagellin, Pam³CSK4 and curdlane. We induced cell death using LPS/ATP and measured real-time cell death using IncuCyte SX5 cell-live analysis.</p></div><div><h3>Results</h3><p>Immunophenotypes of MPI and mAM were comparable despite lower SiglecF and CD₁₁c expression in MPI or mAM after expansion compared to freshly isolated mAM. MPI, mAM and hAM presented similar inflammatory signature but differences in the amplitude of response (exception for Flagellin that induced IL-1-β response in murine models but not in hAM). Overall, MPI had a more pro-inflammatory profile than mAM, as illustrated by a 10-fold production of IL-6 in response to LPS compared to mAM. Differences were observed regarding cell death in mice and human AM models: pyroptotic events were more rapid (1<!--> <!-->h) and important in murine AM models compared to hAM.</p></div><div><h3>Conclusion</h3><p>While clarifying the limits of MPI and <em>ex vivo</em> mAM models, our work showcased consistent inflammatory responses across AM models, albeit with differing response amplitudes. Conversely, mouse and human models exhibited variations in cell death kinetics.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 211"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of hypoxia in Cystic Fibrosis bronchial epithelial cells: Focus on CFTR and TRPA1 channels","authors":"K. Pascarel,&nbsp;J. Colas,&nbsp;T. Carrez,&nbsp;S. Mirval,&nbsp;C. Barrault,&nbsp;F. Becq,&nbsp;C. Vandebrouck","doi":"10.1016/j.rmr.2024.01.089","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.089","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;Cystic Fibrosis (CF) is an autosomal and recessive disease caused by the mutation of a gene located on the chromosome 7: &lt;em&gt;CFTR&lt;/em&gt; (Cystic Fibrosis Transmembrane conductance Regulator). Its codes for CFTR, a protein which plays a role in mucus homeostasis by transporting both chloride ions and water. The most common mutation F508del-CFTR, leads to the absence and malfunction of CFTR at the surface of epithelial cells in various organs especially in lungs. It results in the loss of the mucus clearance properties in the airways, which will cause the obstruction of bronchi and alveola over time. The oxygen (O&lt;sub&gt;2&lt;/sub&gt;) delivery, crucial for aerobic metabolism, is less effective especially for the lung's epithelial cells whose environment is gradually becoming hypoxic. The inability of lungs to realise haematosis, at tissue level is commonly named the respiratory failure.&lt;/p&gt;&lt;p&gt;Our project aims to characterise the impact of hypoxia on ion channels, especially CFTR and TRPA1 (an oxygen sensible calcium channel).&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;p&gt;Cystic Fibrosis Bronchial Epithelial (CFBE) cells-wt (WT) and CFBE F508del (DF) are cultivated in a controlled hypoxic atmosphere (1% O&lt;sub&gt;2&lt;/sub&gt;). Protein expression and quantification have been realised by western blot. CFTR activity have been measured by automated patch-clamp (whole cell recording, WCR) and Ussing chamber while the activity of TRPA1 have been recorded using the Fluo4-AM probe. TRPA1 localisation has been studied by immunostaining.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;p&gt;Here, we show that the change from normoxia (21% O&lt;sub&gt;2&lt;/sub&gt;) to hypoxia (1% O&lt;sub&gt;2&lt;/sub&gt;) is able to induce a fast cellular response with the accumulation of HIF-1α (Hypoxia Inducible Factor) in only 6&lt;!--&gt; &lt;!--&gt;hours in CFBE WT, CFBE-DF and CFBE-DF corrected by the tri-therapy Kaftrio® (Elexacaftor/Tezacaftor/Ivacaftor, ETI). We also observed, that HIF-1α accumulated is reduced in non-corrected CFBE-F508del. Regarding CFTR, our results shows that only F508del-CFTR is impacted by hypoxia at protein level and activity, despite the correction by ETI. Automated WCR patch-clamp and Ussing chamber recordings both show that Kaftrio® corrected F508del-CFTR activity decreases 24&lt;!--&gt; &lt;!--&gt;hours after hypoxia induction. F508del-CFTR currents are decreased by 43% in whole cell configuration while short-circuit current (Isc) CFTR dependent are diminished by around 48%. Concerning TRPA1, hypoxia does not impact the protein accumulation but instead decrease the channel activity by 49% in CFBE-wt, 40% in CFBE-F508del non corrected and 30% when corrected by ETI. Finally, it seems that hypoxia plays a role in TRPA1 location inducing its relocation close to the plasma membrane.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;p&gt;Our data show a reduced amount of CFTR protein accumulated in CFBE F508del corrected or not, which was not observed on the WT form. Electrophysiologic assays show a clear impact of hypoxia on F508de","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 226"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effet de l’absence de microbiote sur l’infection pulmonaire par le virus respiratoire syncytial chez le souriceau","authors":"C. Chottin ,&nbsp;C. Ferret ,&nbsp;C. Maudet ,&nbsp;V. Saint-Criq ,&nbsp;S. Riffault ,&nbsp;D. Descamps","doi":"10.1016/j.rmr.2024.01.047","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.047","url":null,"abstract":"<div><h3>Introduction</h3><p>La bronchiolite est la principale infection respiratoire chez le nourrisson, dont l’agent étiologique principal est le virus respiratoire syncytial (VRS) contre lequel il n’existe pas de vaccins. La sensibilité du nourrisson à l’infection par le VRS est intrinsèquement liée aux caractéristiques de la muqueuse pulmonaire en période périnatale qui évoluent avec la mise en place de l’immunité et la colonisation du poumon par un microbiote bactérien. Bien que décrit pour le microbiote intestinal, le rôle du microbiote pulmonaire sur la maturation de l’immunité de la muqueuse pulmonaire, et donc sur la trajectoire de la santé respiratoire reste peu décrit. Notre étude vise à évaluer si l’absence de microbiote en période périnatale modifie la réplication virale et la réponse immunitaire pulmonaire déclenchée par l’infection VRS. La compréhension des interactions précoces entre le microbiote et la muqueuse pulmonaire pourrait contribuer au développement de nouvelles approches visant à renforcer la réponse immunitaire et ainsi limiter la sévérité de l’infection par le VRS en période néonatale.</p></div><div><h3>Méthodes</h3><p>(1) Infection de souriceaux C57BL/6 dépourvus de flore (animaux axéniques) ou issus de géniteurs colonisés par une flore bactérienne (animaux colonisés) par une souche de VRS recombinant exprimant la luciférase (VRS-Luc).</p><p>(2) Mesure du niveau de réplication virale (bioluminescence et qPCR) et de la réponse immunitaire pulmonaire (infiltration cellulaire dans les lavages bronchoalvéolaires [LBA] et qPCR sur les gènes dépendants des interférons de type 1 [ISG]) à j₁, j₂ et j₄ post-infection.</p></div><div><h3>Résultats</h3><p>Les animaux axéniques montrent une augmentation du niveau de réplication virale entre j₁ et j₄ post-infection par détection de bioluminescence et par qPCR. À l’inverse, les animaux colonisés présentent une diminution du signal entre j₁ et j₄ post-infection. L’infiltration cellulaire dans les LBA des animaux axéniques est augmentée par rapport à celle quantifiée dans les animaux colonisés. L’expression des ISG (IRF7, ISG15 ou OAS) analysée par qPCR est augmentée à j₄ post-infection chez les animaux colonisés, alors que celle des animaux colonisés est induite dès j₁ post-infection.</p></div><div><h3>Conclusion</h3><p>L’absence de flore bactérienne s’accompagne de différences chez le souriceau dans la cinétique de réplication virale et dans la mise en place de la réponse immunitaire lors d’une infection par le VRS. La caractérisation des cellules immunitaires de la muqueuse pulmonaire est prévue afin d’identifier un partenaire cellulaire important dans la défense antivirale influencé par la présence d’un microbiote en période néonatale.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 204"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Surdose en cannabinoïdes de synthèse : attention aux complications respiratoires aiguës","authors":"M. Underner ,&nbsp;J. Perriot ,&nbsp;G. Peiffer ,&nbsp;N. Jaafari ,&nbsp;T. Urban","doi":"10.1016/j.rmr.2024.02.005","DOIUrl":"10.1016/j.rmr.2024.02.005","url":null,"abstract":"","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 262-263"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140068639","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}