Pub Date : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1205
J. Bentley, H. Breckenridge, J. Kreger, A. Goldsmith, C. Lumeng, M. Hershenson
{"title":"Rhinovirus-A1B Decreases Coronavirus NL63 Replication and Inflammation in Adult Mice","authors":"J. Bentley, H. Breckenridge, J. Kreger, A. Goldsmith, C. Lumeng, M. Hershenson","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1205","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1205","url":null,"abstract":"","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"36 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129346594","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1217
M. Longmire, J. Bodnar, J. Pantaleón García, Y. Wang, S. Evans
{"title":"Combined Toll-Like Receptor Agonist Treatment Leads to Activation of Interferon Regulatory Factor 3 in Lung Epithelial Cells In Vitro","authors":"M. Longmire, J. Bodnar, J. Pantaleón García, Y. Wang, S. Evans","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1217","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1217","url":null,"abstract":"","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"46 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121308392","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1213
Y. Palacios, L. Ramón-Luing, A. Ruiz, A. García, A. Sánchez-Monciváis, O. Barreto-Rodríguez, K. Medina-Quero, I. Buendía-Roldán, L. Chávez-Galán
{"title":"The Combined Action of Tumor Necrosis Factor and Gamma Interferon Leads to Inflammation-Cell Death Axis and Senescence in Covid-19 Patients","authors":"Y. Palacios, L. Ramón-Luing, A. Ruiz, A. García, A. Sánchez-Monciváis, O. Barreto-Rodríguez, K. Medina-Quero, I. Buendía-Roldán, L. Chávez-Galán","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1213","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1213","url":null,"abstract":"","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"104 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132986432","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1196
M. Numata-Nakamura, R. Bowen, D. Voelker
Background: A novel coronavirus (SARS-CoV-2) has led to the worldwide spread of pandemic proportions and currently no effective therapy is available. The minor pulmonary surfactant lipids, palmitoyl-oleoyl-phosphatidylglycerol (POPG), and phosphatidylinositol (PI), are potent regulators of inflammatory processes, and are effective as anti-viral agents for multiple respiratory viruses including Respiratory syncytial virus (RSV), Influenza A virus (IAV) and Rhinoviruses (RVs). Objective: The primary objectives of this study are to determine whether POPG or PI are potent against SARS-CoV-2 in vitro, using human airway epithelial cells, and examine the potency of PI against SARS-CoV-2 in vivo, in a hamster model. Methods: We examined efficacies of POPG or PI against SARS-CoV-2 (USA WA/2020) in human bronchial epithelial cells, and nasal epithelial cells from healthy control subjects differentiated by ALI cultures. We quantified SARS-CoV-2 replication by quantitative plaque assays and qRT-PCR. We determined the potency of PI against SARS-CoV- 2 in golden Syrian hamster as in vivo model for SARS-CoV-2 infection. Results: We examined the efficacies of POPG and PI using primary human tracheal and nasal epithelial cells, differentiated in ALI culture. Cells were treated with POPG (10mg/ml) and PI (4mg/ml) added to apical media alone for 16hrs. Subsequently, cells were infected with SARS-CoV-2 at m.o.i = 0.02, for 48hrs, harvested for RNA extraction and qRT-PCR. SARS-CoV-2 replicated in tracheal cells with a 106-fold increase in mRNA. POPG and PI reduced viral mRNA expression by 70% and 85%, respectively (subject numbers n=3). In nasal epithelia, SARS-CoV-2 mRNA expression increased 105 -fold compared to sham infected cultures. Both POPG and PI attenuated the increase in viral mRNA expression by 70% - 82% (subject numbers n=6). We determined the PI effect in an in vivo study in hamsters. Hamsters were challenged with 103 pfu of SARS-CoV-2, either with, or without PI (2mg/hamster) administered intranasally. Hamsters were harvested at Day 3, and lungs were processed for histopathology. Pharyngeal swabs were used to examine viral burden by plaque assays. PI reduced plaque numbers compared to viral infection alone groups at day1 (Virus alone: 2.4±2.7(X104pfu/ml), Virus+PI: 0.9±2.1(X106pfu/ml), p<0.05). PI reduced lung histopathology score at day 3 (Virus alone: 28.0±15.6, Virus+PI: 6.7±7.0, p<0.05). Conclusions: POPG and PI significantly reduced SARS-CoV2 replication in human differentiated airway epithelial cells. PI inhibited SARS-CoV-2 infection in vivo in hamsters. These findings suggest that inhalation of POPG, or PI might be effective as novel anti-viral compounds for treating and preventing SARSCoV- 2 infection.
{"title":"Pulmonary Surfactant Phospholipids as Novel Anti-Virals Against SARS-CoV-2 Infection","authors":"M. Numata-Nakamura, R. Bowen, D. Voelker","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1196","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1196","url":null,"abstract":"Background: A novel coronavirus (SARS-CoV-2) has led to the worldwide spread of pandemic proportions and currently no effective therapy is available. The minor pulmonary surfactant lipids, palmitoyl-oleoyl-phosphatidylglycerol (POPG), and phosphatidylinositol (PI), are potent regulators of inflammatory processes, and are effective as anti-viral agents for multiple respiratory viruses including Respiratory syncytial virus (RSV), Influenza A virus (IAV) and Rhinoviruses (RVs). Objective: The primary objectives of this study are to determine whether POPG or PI are potent against SARS-CoV-2 in vitro, using human airway epithelial cells, and examine the potency of PI against SARS-CoV-2 in vivo, in a hamster model. Methods: We examined efficacies of POPG or PI against SARS-CoV-2 (USA WA/2020) in human bronchial epithelial cells, and nasal epithelial cells from healthy control subjects differentiated by ALI cultures. We quantified SARS-CoV-2 replication by quantitative plaque assays and qRT-PCR. We determined the potency of PI against SARS-CoV- 2 in golden Syrian hamster as in vivo model for SARS-CoV-2 infection. Results: We examined the efficacies of POPG and PI using primary human tracheal and nasal epithelial cells, differentiated in ALI culture. Cells were treated with POPG (10mg/ml) and PI (4mg/ml) added to apical media alone for 16hrs. Subsequently, cells were infected with SARS-CoV-2 at m.o.i = 0.02, for 48hrs, harvested for RNA extraction and qRT-PCR. SARS-CoV-2 replicated in tracheal cells with a 106-fold increase in mRNA. POPG and PI reduced viral mRNA expression by 70% and 85%, respectively (subject numbers n=3). In nasal epithelia, SARS-CoV-2 mRNA expression increased 105 -fold compared to sham infected cultures. Both POPG and PI attenuated the increase in viral mRNA expression by 70% - 82% (subject numbers n=6). We determined the PI effect in an in vivo study in hamsters. Hamsters were challenged with 103 pfu of SARS-CoV-2, either with, or without PI (2mg/hamster) administered intranasally. Hamsters were harvested at Day 3, and lungs were processed for histopathology. Pharyngeal swabs were used to examine viral burden by plaque assays. PI reduced plaque numbers compared to viral infection alone groups at day1 (Virus alone: 2.4±2.7(X104pfu/ml), Virus+PI: 0.9±2.1(X106pfu/ml), p<0.05). PI reduced lung histopathology score at day 3 (Virus alone: 28.0±15.6, Virus+PI: 6.7±7.0, p<0.05). Conclusions: POPG and PI significantly reduced SARS-CoV2 replication in human differentiated airway epithelial cells. PI inhibited SARS-CoV-2 infection in vivo in hamsters. These findings suggest that inhalation of POPG, or PI might be effective as novel anti-viral compounds for treating and preventing SARSCoV- 2 infection.","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"10 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124222584","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1197
S. Rollins, A. Hume, C. Simone-Roach, Y. Tang, C. Chin, D.-Y. Chen, M. Prakash, F. Hawkins, T. Schlaeger, B. Raby, M. Saeed, D. Kotton, E. Mühlberger, R. Wang
Rationale There is a lack of knowledge of how CFTR-deficient airway epithelium intrinsically responds to SARS-CoV-2. Though prior work has demonstrated altered CF airway expression of viral entry factors, it is unknown whether these alterations are protective and whether they reflect host genetic variation or secondary response of chronic inflammation. We address this gap by infecting induced pluripotent stem cell (iPSC)-derived airways from CF patients and syngeneic CFTR-corrected controls with SARS-CoV-2 and assessing differential susceptibility to infection and inflammatory and anti-viral response. MethodsCF (F508del homozygous) and syngeneic CFTR-corrected (CRISPR-Cas9) iPSC- were differentiated into airway epithelium cultured at airliquid interface (ALI) by a directed differentiation protocol that generates a pure population of major and rare airway cell-types. After 21 days in ALI culture, the iPSC-airway were infected with either mock or SARS-CoV-2 (isolate USA-WA1/2020) with MOI of 4, and harvested at 0, 1, 3 days post infection (dpi) for RT-PCR and immune-stainingResultsBoth CF and CFTR-corrected iPSC-airway express viral entry factors of ACE2 and TMPRSS2, and are permissive to SARS-CoV-2 infection. CF iPSC-airway exhibited significantly increase in SARS-CoV-2 nucleocapsid protein (N) transcript at 1 dpi, accompanied by increases in IFN2, RSAD2, and CXCL10 at 3 dpi, compared to its CFTR-corrected counter-part. There are no baseline significant differences in ACE2, TMPRSS2, TP63, NGFR, MUC5B, MUC5AC, SCGB1A1, FOXJ1, FOXI1 expression between CF and CFTR-corrected iPSC-airway before SARS-CoV-2 infection. ConclusionsOur preliminary studies indicate increased early SARS-CoV-2 infection in CFTR-deficient epithelium with accompanied subsequent rise in anti-viral and inflammatory response compared to its genetically controlled CFTR-corrected counterpart. Future studies are aimed at assessing differential CF epithelial kinetics of SARS-CoV-2 viral entry and replication, morphological changes, global transcriptomic response, and how treatment with CFTRmodulator would alter the epithelial response. Ultimately, we aim to establish a reductionist, physiologically relevant model system that is coupled with gene-editing technology to study intrinsic CF epithelial response to SARS-CoV-2, which would generate insights to aid practice guidelines for CF patients, and open future directions to evaluate gene-specific mechanisms of airway response to pathogens. (Figure Presented).
{"title":"Induced Pluripotent Stem Cell Derived Airway to Assess Cystic Fibrosis Intrinsic Epithelial Response to SARS-CoV-2​","authors":"S. Rollins, A. Hume, C. Simone-Roach, Y. Tang, C. Chin, D.-Y. Chen, M. Prakash, F. Hawkins, T. Schlaeger, B. Raby, M. Saeed, D. Kotton, E. Mühlberger, R. Wang","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1197","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1197","url":null,"abstract":"Rationale There is a lack of knowledge of how CFTR-deficient airway epithelium intrinsically responds to SARS-CoV-2. Though prior work has demonstrated altered CF airway expression of viral entry factors, it is unknown whether these alterations are protective and whether they reflect host genetic variation or secondary response of chronic inflammation. We address this gap by infecting induced pluripotent stem cell (iPSC)-derived airways from CF patients and syngeneic CFTR-corrected controls with SARS-CoV-2 and assessing differential susceptibility to infection and inflammatory and anti-viral response. MethodsCF (F508del homozygous) and syngeneic CFTR-corrected (CRISPR-Cas9) iPSC- were differentiated into airway epithelium cultured at airliquid interface (ALI) by a directed differentiation protocol that generates a pure population of major and rare airway cell-types. After 21 days in ALI culture, the iPSC-airway were infected with either mock or SARS-CoV-2 (isolate USA-WA1/2020) with MOI of 4, and harvested at 0, 1, 3 days post infection (dpi) for RT-PCR and immune-stainingResultsBoth CF and CFTR-corrected iPSC-airway express viral entry factors of ACE2 and TMPRSS2, and are permissive to SARS-CoV-2 infection. CF iPSC-airway exhibited significantly increase in SARS-CoV-2 nucleocapsid protein (N) transcript at 1 dpi, accompanied by increases in IFN2, RSAD2, and CXCL10 at 3 dpi, compared to its CFTR-corrected counter-part. There are no baseline significant differences in ACE2, TMPRSS2, TP63, NGFR, MUC5B, MUC5AC, SCGB1A1, FOXJ1, FOXI1 expression between CF and CFTR-corrected iPSC-airway before SARS-CoV-2 infection. ConclusionsOur preliminary studies indicate increased early SARS-CoV-2 infection in CFTR-deficient epithelium with accompanied subsequent rise in anti-viral and inflammatory response compared to its genetically controlled CFTR-corrected counterpart. Future studies are aimed at assessing differential CF epithelial kinetics of SARS-CoV-2 viral entry and replication, morphological changes, global transcriptomic response, and how treatment with CFTRmodulator would alter the epithelial response. Ultimately, we aim to establish a reductionist, physiologically relevant model system that is coupled with gene-editing technology to study intrinsic CF epithelial response to SARS-CoV-2, which would generate insights to aid practice guidelines for CF patients, and open future directions to evaluate gene-specific mechanisms of airway response to pathogens. (Figure Presented).","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"15 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123624409","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1202
J. Campos-Gómez, Q. Li, S. S. Hussain, J.E. Peabody Lever, C. Fernandez Petty, H. Kim, S. Rowe
{"title":"Mucociliary Transport Augmenting Agents Block SARS-Cov-2 Replication in Human Bronchial Epithelial Cells In Vitro","authors":"J. Campos-Gómez, Q. Li, S. S. Hussain, J.E. Peabody Lever, C. Fernandez Petty, H. Kim, S. Rowe","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1202","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1202","url":null,"abstract":"","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128281588","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1206
K. Rubey, A. J. Paris, A. Mukhitov, S. Lin, J. Myerson, J. Brenner, G. Worthen, V. Krymskaya, J. Heimall
{"title":"Neutrophil-Avid Nanocarriers, a Future Potent Autosomal Dominant Hyper Ig-E Syndrome STAT3 Deficiency Therapeutic","authors":"K. Rubey, A. J. Paris, A. Mukhitov, S. Lin, J. Myerson, J. Brenner, G. Worthen, V. Krymskaya, J. Heimall","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1206","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1206","url":null,"abstract":"","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"9 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134538183","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1214
G. Gupta, S.-H. Kim, H. Shin, L. Wei, I. Rosas, N. Kaminski, L. Sharma, C. D. Dela Cruz, Min-Jong Kang
{"title":"Cigarette Smoke Exposure or Nigericin/LPS Stimulation Induces MAVS Aggregation, Which Is Further Augmented in NLRX1 Deficiency","authors":"G. Gupta, S.-H. Kim, H. Shin, L. Wei, I. Rosas, N. Kaminski, L. Sharma, C. D. Dela Cruz, Min-Jong Kang","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1214","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1214","url":null,"abstract":"","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"108 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123172084","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1212
K. Shepardson, J. Roemer, A. Rynda-Apple
RATIONALE: Recently, there has been an increased incidence of invasive pulmonary aspergillosis (IPA), caused by the human fungal pathogen Aspergillus fumigatus (Af), occurring in patients infected with influenza or SARS-CoV-2. Along with the recently described involvement of type I interferon (IFN) signaling in increased Af susceptibility during viral infection in mice, this strongly indicates that anti-viral immune responses, such as type I IFNs, create an environment permissive to fungal infection. Supporting this, we found that type I IFN signaling, via the type I IFN receptor 2 (IFNAR2) of IFNAR1/2, contributes to regulation of susceptibility to and damage from influenza in mice, while others have found that IFNAR2 expression correlates with SARS-CoV-2 infection severity. As clinical outcome to Af is associated with host tissue damage, this suggests that IFNAR2's regulation of damage response during pulmonary infection may control the immune status of the lung, via tissue damage, allowing for fungal infection to occur. METHODS: We are utilizing a murine pulmonary infection model, to identify distinct roles for IFNAR2 and IFNAR1 and type I IFN signaling in regulating both damage and clearance during IPA. We employed proteomic, histological, and molecular approaches to determine the components and extent of the damage response. RESULTS: We found that absence of IFNAR2 (Ifnar2-/- mice) resulted in increased damage, weight loss, and morbidity early during Af infection compared to WT and Ifnar1-/- mice. Additionally, we also found that both WT and Ifnar1-/- mice had decreased Af clearance early during infection compared to Ifnar2-/- mice and that this difference in killing of Af required in vivo interactions/signaling. However, as Af infection progressed we found that although Ifnar2-/- mice cleared Af early, this did not prevent invasive hyphal growth from occurring. This invasive growth in the Ifnar2-/- mice was found to be associated with increased damage and cell death in the Af lesions within the lung. Importantly, our results suggest that this IFNAR2 damage response is being mediated by distinct type I IFNs, specifically IFNβ. CONCLUSIONS: Together, our results begin to establish a role for IFNAR2 in regulation of the host damage response to Af and suggests that the type of type I IFN signaling may contribute to a permissive environment allowing for Af infection to occur. Understanding the mechanisms involved in IFNAR regulation of damage and anti-fungal immunity could inform design of better treatments aimed at minimizing damage in patients with IPA or controlling pulmonary tissue damage.
{"title":"Type I IFN Signaling Controls Damage and Clearance During Pulmonary Aspergillus Fumigatus Infection","authors":"K. Shepardson, J. Roemer, A. Rynda-Apple","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1212","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1212","url":null,"abstract":"RATIONALE: Recently, there has been an increased incidence of invasive pulmonary aspergillosis (IPA), caused by the human fungal pathogen Aspergillus fumigatus (Af), occurring in patients infected with influenza or SARS-CoV-2. Along with the recently described involvement of type I interferon (IFN) signaling in increased Af susceptibility during viral infection in mice, this strongly indicates that anti-viral immune responses, such as type I IFNs, create an environment permissive to fungal infection. Supporting this, we found that type I IFN signaling, via the type I IFN receptor 2 (IFNAR2) of IFNAR1/2, contributes to regulation of susceptibility to and damage from influenza in mice, while others have found that IFNAR2 expression correlates with SARS-CoV-2 infection severity. As clinical outcome to Af is associated with host tissue damage, this suggests that IFNAR2's regulation of damage response during pulmonary infection may control the immune status of the lung, via tissue damage, allowing for fungal infection to occur. METHODS: We are utilizing a murine pulmonary infection model, to identify distinct roles for IFNAR2 and IFNAR1 and type I IFN signaling in regulating both damage and clearance during IPA. We employed proteomic, histological, and molecular approaches to determine the components and extent of the damage response. RESULTS: We found that absence of IFNAR2 (Ifnar2-/- mice) resulted in increased damage, weight loss, and morbidity early during Af infection compared to WT and Ifnar1-/- mice. Additionally, we also found that both WT and Ifnar1-/- mice had decreased Af clearance early during infection compared to Ifnar2-/- mice and that this difference in killing of Af required in vivo interactions/signaling. However, as Af infection progressed we found that although Ifnar2-/- mice cleared Af early, this did not prevent invasive hyphal growth from occurring. This invasive growth in the Ifnar2-/- mice was found to be associated with increased damage and cell death in the Af lesions within the lung. Importantly, our results suggest that this IFNAR2 damage response is being mediated by distinct type I IFNs, specifically IFNβ. CONCLUSIONS: Together, our results begin to establish a role for IFNAR2 in regulation of the host damage response to Af and suggests that the type of type I IFN signaling may contribute to a permissive environment allowing for Af infection to occur. Understanding the mechanisms involved in IFNAR regulation of damage and anti-fungal immunity could inform design of better treatments aimed at minimizing damage in patients with IPA or controlling pulmonary tissue damage.","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"12 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125289020","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 : 2022-05-01DOI: 10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1193
N. Bartlett, T. Williams, S. Loo, J. Girkin
Rationale We have previously reported blocking the IL-25 receptor (IL-17RB) prevented viral increased allergic airways inflammation and this was associated with reduced lung viral load. To investigate IL-25 regulation of airway anti-viral immunity we hypothesised that IL-25 directly inhibits airway epithelial cell (AEC) type I/III interferon expression and antibody blockade of IL-25 in vivo boosts lung interferon expression and reduces lung viral load in parallel with reduced type 2 airway inflammation. Methods In vitro Immunofluorescence was used to visualise epithelial IL-25 and IL- 17RB proteins in endobronchial biopsies from patients with asthma and healthy subjects and in AEC differentiated at ALI. AEC from n = 14 donors with asthma were differentiated at the air-liquid interface (ALI) and infected with RV-A1, MOI=0.1. A subset of AECs was treated with anti-IL-25 mAb (LNR125) before infecting with RV-A1 or human coronavirus 229E. Differentiated AEC from healthy donors were treated with recombinant IL-25 protein and infected with RV-A1. Nanostring immune transcriptomic data expressed as digital mRNA counts for exact copy number or was expressed as log2 fold change ratio against -log10 Bejamini-Yekutieli-corrected p-values. In vivo 6- 8-week-old, BALB/c mice sensitised and intranasally challenged daily for 3 days with ovalbumin to induced allergic airways disease. A single subcutaneous injection of 250 μg LNR125 was administered during ovalbumin challenge. Mice were then infected i.n. with RV-A1, 6 hours after final allergen challenge. On day 1 and day 7 post-infection, BAL were collected, lung lobe tissue was collected for viral RNA and cytokine expression. Results IL-25 and IL-17RB were constitutively expressed at the apical surface of airway epithelium in biopsies and AEC cultures. RV infection increased IL-25 expression by AEC from asthmatic donors. LNR125 treatment reduced IL-25 mRNA and significantly increased RV induced IFN-β a and IFN-λ protein expression and this was confirmed by Nanostring transcriptomic analyses which also identified down-regulated type-2 immune genes CCL26 (eotaxin 3) and IL1RL1(IL-33 receptor). LN125 treatment also increased IFN-λ expression by 229E-infected differentiated AECs. IL-25 treatment increased viral load associated with 50% reduced expression of IFN-β and CXCL10 and 75% reduced IFN-λ. Allergen challenged, RV-infected mice treated with LNR125 had significantly increased BAL IFN-β protein and 60% reduction in lung viral load associated with reduced IL-25, IL-4, IL-5 and IL-13 BAL proteins compared to controls. Conclusion IL-25-induced inflammation combined with suppression of AEC anti-viral immunity identify IL-25 as a central mediator of viral asthma exacerbations and therefore a target for mAb-based treatment.
{"title":"IL-25 Inhibits Airway Anti-Viral Immunity and Promotes Virus Exacerbation of Allergic Airways Disease","authors":"N. Bartlett, T. Williams, S. Loo, J. Girkin","doi":"10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1193","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1193","url":null,"abstract":"Rationale We have previously reported blocking the IL-25 receptor (IL-17RB) prevented viral increased allergic airways inflammation and this was associated with reduced lung viral load. To investigate IL-25 regulation of airway anti-viral immunity we hypothesised that IL-25 directly inhibits airway epithelial cell (AEC) type I/III interferon expression and antibody blockade of IL-25 in vivo boosts lung interferon expression and reduces lung viral load in parallel with reduced type 2 airway inflammation. Methods In vitro Immunofluorescence was used to visualise epithelial IL-25 and IL- 17RB proteins in endobronchial biopsies from patients with asthma and healthy subjects and in AEC differentiated at ALI. AEC from n = 14 donors with asthma were differentiated at the air-liquid interface (ALI) and infected with RV-A1, MOI=0.1. A subset of AECs was treated with anti-IL-25 mAb (LNR125) before infecting with RV-A1 or human coronavirus 229E. Differentiated AEC from healthy donors were treated with recombinant IL-25 protein and infected with RV-A1. Nanostring immune transcriptomic data expressed as digital mRNA counts for exact copy number or was expressed as log2 fold change ratio against -log10 Bejamini-Yekutieli-corrected p-values. In vivo 6- 8-week-old, BALB/c mice sensitised and intranasally challenged daily for 3 days with ovalbumin to induced allergic airways disease. A single subcutaneous injection of 250 μg LNR125 was administered during ovalbumin challenge. Mice were then infected i.n. with RV-A1, 6 hours after final allergen challenge. On day 1 and day 7 post-infection, BAL were collected, lung lobe tissue was collected for viral RNA and cytokine expression. Results IL-25 and IL-17RB were constitutively expressed at the apical surface of airway epithelium in biopsies and AEC cultures. RV infection increased IL-25 expression by AEC from asthmatic donors. LNR125 treatment reduced IL-25 mRNA and significantly increased RV induced IFN-β a and IFN-λ protein expression and this was confirmed by Nanostring transcriptomic analyses which also identified down-regulated type-2 immune genes CCL26 (eotaxin 3) and IL1RL1(IL-33 receptor). LN125 treatment also increased IFN-λ expression by 229E-infected differentiated AECs. IL-25 treatment increased viral load associated with 50% reduced expression of IFN-β and CXCL10 and 75% reduced IFN-λ. Allergen challenged, RV-infected mice treated with LNR125 had significantly increased BAL IFN-β protein and 60% reduction in lung viral load associated with reduced IL-25, IL-4, IL-5 and IL-13 BAL proteins compared to controls. Conclusion IL-25-induced inflammation combined with suppression of AEC anti-viral immunity identify IL-25 as a central mediator of viral asthma exacerbations and therefore a target for mAb-based treatment.","PeriodicalId":307658,"journal":{"name":"A28. THE HOST DELIVERS","volume":"10 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125770540","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}
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