Pub Date : 2020-05-01DOI: 10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2964
J. Londino, J. Adair, R. K. Mallampalli
{"title":"Interferon Lambda Signaling in Macrophages Is Necessary for the Antiviral Response to Influenza","authors":"J. Londino, J. Adair, R. K. Mallampalli","doi":"10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2964","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2964","url":null,"abstract":"","PeriodicalId":188042,"journal":{"name":"B29. INFECTION AND IMMUNE INTERPLAY IN LUNG INJURY","volume":"41 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141208652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-05-01DOI: 10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2978
Timothy Wang, Jerilyn Gray, A. Nadeem, I. Lang, W. Zacharias, H. Deshmukh
We demonstrated that disruption of intestinal commensal bacteria with antibiotics (ABX) severely diminished the levels of cytokine interleukin (IL)-22 in the newborn lung and rendered the ABX-exposed newborn mice susceptible post challenge with Streptococcus pneumoniae (S. pneumoniae). These experimental data suggest that colonization by intestinal commensal bacteria is vital for newborn’s lung defense against respiratory pathogens. Pregnant mouse dams were treated with ABX, and newborn mice were challenged with intratracheal S. pneumoniae (on postnatal day 5). Lungs were harvested and stained. AT2 cells and fibroblast harvested from ABX-treated or ABX-free newborn mice were cultured in matrigel to generate alveolar organoids. After harvest, the organoids were stained. To test the role of IL-22 signaling, Il22rafl/fl mice were bred with SftpcCreERT2 mice. The progeny was treated with tamoxifen and then challenged with S. pneumoniae. We found increased alveolar injury, loss of AT1 cells and reduced the frequency of proliferating AT2 cells in ABX-treated newborn mice compared to ABX-free newborn mice post-challenge with S. pneumoniae. Reconstitution of intestinal commensal bacteria or treatment with recombinant IL-22 restored the numbers of proliferating AT2 cells, reduced the injury score, and improved survival in ABX-treated newborn mice. Organoids derived ABX-exposed newborns demonstrated reduced growth and diminished AT1 differentiation. Using a developmentally appropriate and clinically relevant model, we report that intestinal commensal bacteria promote AT2 self-renewal and AT1 differentiation in an IL-22 dependent fashion, thus preparing the newborn to fight potentially fatal respiratory pathogens.
{"title":"Intestinal Commensal Bacteria Promote AT2 Self-Renewal and AT1 Differentiation in an IL-22 Dependent Fashion and Prepare the Newborn to Fight Potentially Fatal Respiratory Pathogens","authors":"Timothy Wang, Jerilyn Gray, A. Nadeem, I. Lang, W. Zacharias, H. Deshmukh","doi":"10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2978","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2978","url":null,"abstract":"\u0000 We demonstrated that disruption of intestinal commensal bacteria with antibiotics (ABX) severely diminished the levels of cytokine interleukin (IL)-22 in the newborn lung and rendered the ABX-exposed newborn mice susceptible post challenge with Streptococcus pneumoniae (S. pneumoniae). These experimental data suggest that colonization by intestinal commensal bacteria is vital for newborn’s lung defense against respiratory pathogens. Pregnant mouse dams were treated with ABX, and newborn mice were challenged with intratracheal S. pneumoniae (on postnatal day 5). Lungs were harvested and stained. AT2 cells and fibroblast harvested from ABX-treated or ABX-free newborn mice were cultured in matrigel to generate alveolar organoids. After harvest, the organoids were stained. To test the role of IL-22 signaling, Il22rafl/fl mice were bred with SftpcCreERT2 mice. The progeny was treated with tamoxifen and then challenged with S. pneumoniae. We found increased alveolar injury, loss of AT1 cells and reduced the frequency of proliferating AT2 cells in ABX-treated newborn mice compared to ABX-free newborn mice post-challenge with S. pneumoniae. Reconstitution of intestinal commensal bacteria or treatment with recombinant IL-22 restored the numbers of proliferating AT2 cells, reduced the injury score, and improved survival in ABX-treated newborn mice. Organoids derived ABX-exposed newborns demonstrated reduced growth and diminished AT1 differentiation. Using a developmentally appropriate and clinically relevant model, we report that intestinal commensal bacteria promote AT2 self-renewal and AT1 differentiation in an IL-22 dependent fashion, thus preparing the newborn to fight potentially fatal respiratory pathogens.","PeriodicalId":188042,"journal":{"name":"B29. INFECTION AND IMMUNE INTERPLAY IN LUNG INJURY","volume":"8 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133835678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-05-01DOI: 10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2962
P. Estèves, B. Allard, A. Celle, I. Dupin, E. Maurat, O. Ousova, M. Thumerel, T. Lassere, R. Marthan, P. Girodet, P. Berger, T. Trian
Viral infection of the bronchial epithelium (BE), particularly with rhinovirus (RV) has been implicated in the vast majority of exacerbations in severe asthma. Preventing these exacerbations remains a major unmet need. Bronchial remodeling in severe asthma has been characterized by increased bronchial smooth muscle (BSM) mass, which had a poor prognostic value, since higher BSM mass was associated with increased exacerbation rate. However, the role of asthmatic BSM in RV infections of the BE has never been demonstrated. We thus hypothesized that asthmatic BSM increased both BE susceptibility and response to RV infection. We then enrolled 19 severe asthmatic patients from the COBRA cohort, and 37 control subjects. We designed cell biologic, transcriptomic and proteomic approaches both in vitro using coculture model of BE with BSM cells and ex vivo using patients’ biopsies. BE cells were cultured and differentiated in air-liquid interface. RV particle number within BE cells was measured using digital PCR. Proteomic and transcriptomic data were analyzed using IPA (Qiagen). Cytokines concentrations in the supernatant were assessed using ELISA assays. Finally, PkR pathway was analyzed by western blotting. Severe asthmatic patients were similar to control subjects in terms of sex ratio, age and BMI. Not surprisingly asthmatic patients presented a lower FEV-1. We found that asthmatic BSM cells co-cultured with BE cells increased RV replication and level of IL6 in the supernatant compare to that co-cultured with control BSM cells. Large scaled proteomic and transcriptomic analysis highlighted CCL-20 production by asthmatic BSM cells, as a potential candidate to specifically increase the RV infection of the BE cells. Blocking CCL-20 in the co-culture supernatant with asthmatic BSM cells reduced the number of RV particles within the BE cells. Moreover, such a co-culture decreased the PkR pathway activation after RV infection. Indeed, we showed a decreased level of the activated form of PkR and eIF2-alpha. Finally, a direct stimulation of BE cells with CCL-20 reproduced both the increased number of RV particles and the down-regulation of the PkR pathway. In conclusion, these results clearly demonstrated that asthmatic BSM increased the BE response to RV, through an increase of CCL-20 production by BSM, which in turn, down-regulated PKR response of the BE leading to increase RV replication and particles number in BE. To the best of our knowledge, it is the very first time that such a direct bottom-up effect of asthmatic BSM on BE has been demonstrated.
{"title":"Bronchial Smooth Muscle of Severe Asthmatic Patients Decreases Rhinovirus Replication Within the Epithelium Through a CCL-20/PkR/eIF2alpha-Dependent Pathway","authors":"P. Estèves, B. Allard, A. Celle, I. Dupin, E. Maurat, O. Ousova, M. Thumerel, T. Lassere, R. Marthan, P. Girodet, P. Berger, T. Trian","doi":"10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2962","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2962","url":null,"abstract":"Viral infection of the bronchial epithelium (BE), particularly with rhinovirus (RV) has been implicated in the vast majority of exacerbations in severe asthma. Preventing these exacerbations remains a major unmet need. Bronchial remodeling in severe asthma has been characterized by increased bronchial smooth muscle (BSM) mass, which had a poor prognostic value, since higher BSM mass was associated with increased exacerbation rate. However, the role of asthmatic BSM in RV infections of the BE has never been demonstrated. We thus hypothesized that asthmatic BSM increased both BE susceptibility and response to RV infection. We then enrolled 19 severe asthmatic patients from the COBRA cohort, and 37 control subjects. We designed cell biologic, transcriptomic and proteomic approaches both in vitro using coculture model of BE with BSM cells and ex vivo using patients’ biopsies. BE cells were cultured and differentiated in air-liquid interface. RV particle number within BE cells was measured using digital PCR. Proteomic and transcriptomic data were analyzed using IPA (Qiagen). Cytokines concentrations in the supernatant were assessed using ELISA assays. Finally, PkR pathway was analyzed by western blotting. Severe asthmatic patients were similar to control subjects in terms of sex ratio, age and BMI. Not surprisingly asthmatic patients presented a lower FEV-1. We found that asthmatic BSM cells co-cultured with BE cells increased RV replication and level of IL6 in the supernatant compare to that co-cultured with control BSM cells. Large scaled proteomic and transcriptomic analysis highlighted CCL-20 production by asthmatic BSM cells, as a potential candidate to specifically increase the RV infection of the BE cells. Blocking CCL-20 in the co-culture supernatant with asthmatic BSM cells reduced the number of RV particles within the BE cells. Moreover, such a co-culture decreased the PkR pathway activation after RV infection. Indeed, we showed a decreased level of the activated form of PkR and eIF2-alpha. Finally, a direct stimulation of BE cells with CCL-20 reproduced both the increased number of RV particles and the down-regulation of the PkR pathway. In conclusion, these results clearly demonstrated that asthmatic BSM increased the BE response to RV, through an increase of CCL-20 production by BSM, which in turn, down-regulated PKR response of the BE leading to increase RV replication and particles number in BE. To the best of our knowledge, it is the very first time that such a direct bottom-up effect of asthmatic BSM on BE has been demonstrated.","PeriodicalId":188042,"journal":{"name":"B29. INFECTION AND IMMUNE INTERPLAY IN LUNG INJURY","volume":"33 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132607041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：