CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection

BackgroundCigarette smoke (CS) exposure increases the frequency and severity of respiratory influenza A virus (IAV) infections in humans and increases mortality in mice. There is evidence that Cytochrome P450 family 1 subfamily B member 1 (CYP1B1) enhances lung injury during hyperoxic exposure in an...

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Main Authors: Wenxin Wu, Jeremy S. Alexander, J. Leland Booth, Magdalena Chlebicz, Craig A. Miller, Chao Xu, Susan Kovats, Jordan P. Metcalf
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1600025/full
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author Wenxin Wu
Jeremy S. Alexander
J. Leland Booth
Magdalena Chlebicz
Craig A. Miller
Chao Xu
Susan Kovats
Susan Kovats
Jordan P. Metcalf
Jordan P. Metcalf
Jordan P. Metcalf
author_facet Wenxin Wu
Jeremy S. Alexander
J. Leland Booth
Magdalena Chlebicz
Craig A. Miller
Chao Xu
Susan Kovats
Susan Kovats
Jordan P. Metcalf
Jordan P. Metcalf
Jordan P. Metcalf
author_sort Wenxin Wu
collection DOAJ
description BackgroundCigarette smoke (CS) exposure increases the frequency and severity of respiratory influenza A virus (IAV) infections in humans and increases mortality in mice. There is evidence that Cytochrome P450 family 1 subfamily B member 1 (CYP1B1) enhances lung injury during hyperoxic exposure in animal models. We used an in-vivo mouse model to assess the hypothesis that CYP1B1 modifies innate immune responses to CS and alters survival during IAV infection.MethodsWe measured CYP1B1 induction in bronchial epithelial cells in smokers and nonsmokers obtained by bronchoscopy by whole transcriptome analysis. To determine whether CYP1B1 knockout (CYP KO) improves mortality and reduces lung injury in CS-exposed mice, we compared the survival rates, host immune responses, and lung-to-body weight ratio of CYP KO mice with the wild-type (WT) mice following challenge with IAV with or without CS exposure.ResultsCYP1B1 is one of the most highly upregulated genes in human lung epithelia derived from cigarette smokers. In CS-exposed mice, CYP1B1 knockout (CYP KO) significantly increased survival during IAV infection. In both nonsmoking (NS) and CS mice, CYP KO significantly enhanced IAV-induced increases in total immune cell numbers in bronchoalveolar lavage fluid (BALF) without causing additional lung injury. CYP KO caused a more rapid IFN-γ response to IAV infection in lungs from both NS- and CS-exposed mice. Specifically, we found that IFN-γ was significantly increased in BALF and in the lung at day 5 post-infection (p.i.) in these mice. Flow cytometry showed that innate lymphocytes produced early IFN-γ in the lungs of KO mice. We confirmed the importance of IFN-γ for CYP KO survival by adding IFN-γ antibody to IAV-infected CYP KO mice. IFN-γ antibody treatment of CS-exposed CYP KO mice completely abolished the improved survival rate seen in KO mice without IFN-γ antibody treatment. However, IFN-γ administration to CS-exposed WT mice did not increase survival rates as seen in CS-exposed CYP KO mice after IAV infection.ConclusionsOur results demonstrate that CYP KO in mice protects against CS-enhanced susceptibility of smokers during influenza infection. IFN-γ is required but not sufficient for the protection of CS-exposed CYP KO mice against lethal IAV infection.
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spelling doaj-art-88ca7be300224df79a250972ba7bbcf42025-08-20T03:16:08ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.16000251600025CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infectionWenxin Wu0Jeremy S. Alexander1J. Leland Booth2Magdalena Chlebicz3Craig A. Miller4Chao Xu5Susan Kovats6Susan Kovats7Jordan P. Metcalf8Jordan P. Metcalf9Jordan P. Metcalf10Pulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United StatesPulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United StatesPulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United StatesArthritis & Clinical Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, United StatesDepartment of Veterinary Pathobiology, College of Veterinary Medicine, Oklahoma State University, Oklahoma State University, Stillwater, OK, United StatesDepartment of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United StatesArthritis & Clinical Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, United StatesDepartment of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United StatesPulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United StatesDepartment of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United StatesVeterans Affairs Medical Center, Oklahoma City, OK, United StatesBackgroundCigarette smoke (CS) exposure increases the frequency and severity of respiratory influenza A virus (IAV) infections in humans and increases mortality in mice. There is evidence that Cytochrome P450 family 1 subfamily B member 1 (CYP1B1) enhances lung injury during hyperoxic exposure in animal models. We used an in-vivo mouse model to assess the hypothesis that CYP1B1 modifies innate immune responses to CS and alters survival during IAV infection.MethodsWe measured CYP1B1 induction in bronchial epithelial cells in smokers and nonsmokers obtained by bronchoscopy by whole transcriptome analysis. To determine whether CYP1B1 knockout (CYP KO) improves mortality and reduces lung injury in CS-exposed mice, we compared the survival rates, host immune responses, and lung-to-body weight ratio of CYP KO mice with the wild-type (WT) mice following challenge with IAV with or without CS exposure.ResultsCYP1B1 is one of the most highly upregulated genes in human lung epithelia derived from cigarette smokers. In CS-exposed mice, CYP1B1 knockout (CYP KO) significantly increased survival during IAV infection. In both nonsmoking (NS) and CS mice, CYP KO significantly enhanced IAV-induced increases in total immune cell numbers in bronchoalveolar lavage fluid (BALF) without causing additional lung injury. CYP KO caused a more rapid IFN-γ response to IAV infection in lungs from both NS- and CS-exposed mice. Specifically, we found that IFN-γ was significantly increased in BALF and in the lung at day 5 post-infection (p.i.) in these mice. Flow cytometry showed that innate lymphocytes produced early IFN-γ in the lungs of KO mice. We confirmed the importance of IFN-γ for CYP KO survival by adding IFN-γ antibody to IAV-infected CYP KO mice. IFN-γ antibody treatment of CS-exposed CYP KO mice completely abolished the improved survival rate seen in KO mice without IFN-γ antibody treatment. However, IFN-γ administration to CS-exposed WT mice did not increase survival rates as seen in CS-exposed CYP KO mice after IAV infection.ConclusionsOur results demonstrate that CYP KO in mice protects against CS-enhanced susceptibility of smokers during influenza infection. IFN-γ is required but not sufficient for the protection of CS-exposed CYP KO mice against lethal IAV infection.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1600025/fullinfluenza virusCYP1B1innatesmokingIFN-γinflammation
spellingShingle Wenxin Wu
Jeremy S. Alexander
J. Leland Booth
Magdalena Chlebicz
Craig A. Miller
Chao Xu
Susan Kovats
Susan Kovats
Jordan P. Metcalf
Jordan P. Metcalf
Jordan P. Metcalf
CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection
Frontiers in Immunology
influenza virus
CYP1B1
innate
smoking
IFN-γ
inflammation
title CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection
title_full CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection
title_fullStr CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection
title_full_unstemmed CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection
title_short CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection
title_sort cyp1b1 knockout enhanced ifn γ production is required but not sufficient for protection of cigarette smoke exposed mice against lethal influenza virus infection
topic influenza virus
CYP1B1
innate
smoking
IFN-γ
inflammation
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1600025/full
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