Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolytica

Mannheimia haemolytica (M. haemolytica) cause mastitis in sheep, acute sepsis in newborn lambs, and co-infections with various pathogens, leading to bovine respiratory disease syndrome (BRDS), these infections have resulted in significant economic losses to both domestic and international farming in...

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Main Authors: Ke Shang, Yuanji Gao, Jiangbo Du, Chang Liu, Jinglei Dai, Junfeng Zhang, Yanyan Jia, Zuhua Yu, Songbiao Chen, Zhongyu Liu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Cellular and Infection Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcimb.2025.1578027/full
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author Ke Shang
Ke Shang
Ke Shang
Yuanji Gao
Yuanji Gao
Jiangbo Du
Chang Liu
Chang Liu
Jinglei Dai
Jinglei Dai
Junfeng Zhang
Junfeng Zhang
Yanyan Jia
Yanyan Jia
Zuhua Yu
Zuhua Yu
Songbiao Chen
Songbiao Chen
Zhongyu Liu
author_facet Ke Shang
Ke Shang
Ke Shang
Yuanji Gao
Yuanji Gao
Jiangbo Du
Chang Liu
Chang Liu
Jinglei Dai
Jinglei Dai
Junfeng Zhang
Junfeng Zhang
Yanyan Jia
Yanyan Jia
Zuhua Yu
Zuhua Yu
Songbiao Chen
Songbiao Chen
Zhongyu Liu
author_sort Ke Shang
collection DOAJ
description Mannheimia haemolytica (M. haemolytica) cause mastitis in sheep, acute sepsis in newborn lambs, and co-infections with various pathogens, leading to bovine respiratory disease syndrome (BRDS), these infections have resulted in significant economic losses to both domestic and international farming industries. An in-depth understanding of the pathogenic mechanisms of M. haemolytica is crucial for the prevention and control of this disease. Outer membrane vesicles (OMVs) play a vital role in bacterial pathogenesis, serving as key mediators of interactions between Gram-negative bacteria and their hosts. However, the specific role of OMVs in the pathogenic process of M. haemolytica remains poorly understood. To address this, we isolated OMVs from the Mannheimia haemolytica Type A5 strain (MH-5) using ultracentrifugation and subsequently characterized their secretory properties, protein composition, and immunogenicity through electron microscopy, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and cellular experiments. The electron microscopy results indicated that the MH-5 strain secreted OMVs under natural growth conditions. Proteomic and bioinformatics analyses revealed that these OMVs contained 282 proteins, with significant enrichment in proteins related to immunity, iron metabolism, and catalytic activity. Cellular experiments demonstrated that, compared to the control group, the OMVs group exhibited a significant increase in the mRNA expression of IL-1β, IL-6, and TNF-α, with secretion levels increasing in a dose-dependent manner, thereby enhancing the inflammatory response. These findings lay the groundwork for further exploration of the role of OMVs in the pathogenesis of M. haemolytica and provide insights for the development of effective vaccines and antibiotics against this pathogen.
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spelling doaj-art-e73de5cdafa649b2a966373659bfe5ba2025-08-20T02:22:36ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-06-011510.3389/fcimb.2025.15780271578027Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolyticaKe Shang0Ke Shang1Ke Shang2Yuanji Gao3Yuanji Gao4Jiangbo Du5Chang Liu6Chang Liu7Jinglei Dai8Jinglei Dai9Junfeng Zhang10Junfeng Zhang11Yanyan Jia12Yanyan Jia13Zuhua Yu14Zuhua Yu15Songbiao Chen16Songbiao Chen17Zhongyu Liu18The 989th Hospital of the Joint Logistics Support Force of Chinese People’s Liberation Army, Luoyang, ChinaLaboratory of Functional Microbiology and Animal Health, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, ChinaLuoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang, ChinaLaboratory of Functional Microbiology and Animal Health, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, ChinaLuoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang, ChinaThe 989th Hospital of the Joint Logistics Support Force of Chinese People’s Liberation Army, Luoyang, ChinaLaboratory of Functional Microbiology and Animal Health, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, ChinaLuoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang, ChinaLaboratory of Functional Microbiology and Animal Health, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, ChinaLuoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang, ChinaThe 989th Hospital of the Joint Logistics Support Force of Chinese People’s Liberation Army, Luoyang, ChinaCollege of Medical Technology and Engineering, Henan University of Science and Technology, Luoyang, ChinaLaboratory of Functional Microbiology and Animal Health, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, ChinaLuoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang, ChinaLaboratory of Functional Microbiology and Animal Health, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, ChinaLuoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang, ChinaLaboratory of Functional Microbiology and Animal Health, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, ChinaLuoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang, ChinaThe 989th Hospital of the Joint Logistics Support Force of Chinese People’s Liberation Army, Luoyang, ChinaMannheimia haemolytica (M. haemolytica) cause mastitis in sheep, acute sepsis in newborn lambs, and co-infections with various pathogens, leading to bovine respiratory disease syndrome (BRDS), these infections have resulted in significant economic losses to both domestic and international farming industries. An in-depth understanding of the pathogenic mechanisms of M. haemolytica is crucial for the prevention and control of this disease. Outer membrane vesicles (OMVs) play a vital role in bacterial pathogenesis, serving as key mediators of interactions between Gram-negative bacteria and their hosts. However, the specific role of OMVs in the pathogenic process of M. haemolytica remains poorly understood. To address this, we isolated OMVs from the Mannheimia haemolytica Type A5 strain (MH-5) using ultracentrifugation and subsequently characterized their secretory properties, protein composition, and immunogenicity through electron microscopy, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and cellular experiments. The electron microscopy results indicated that the MH-5 strain secreted OMVs under natural growth conditions. Proteomic and bioinformatics analyses revealed that these OMVs contained 282 proteins, with significant enrichment in proteins related to immunity, iron metabolism, and catalytic activity. Cellular experiments demonstrated that, compared to the control group, the OMVs group exhibited a significant increase in the mRNA expression of IL-1β, IL-6, and TNF-α, with secretion levels increasing in a dose-dependent manner, thereby enhancing the inflammatory response. These findings lay the groundwork for further exploration of the role of OMVs in the pathogenesis of M. haemolytica and provide insights for the development of effective vaccines and antibiotics against this pathogen.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1578027/fullMannheimia haemolytica type A5outer membrane vesiclesproteomicsinflammatory responsein vitro test
spellingShingle Ke Shang
Ke Shang
Ke Shang
Yuanji Gao
Yuanji Gao
Jiangbo Du
Chang Liu
Chang Liu
Jinglei Dai
Jinglei Dai
Junfeng Zhang
Junfeng Zhang
Yanyan Jia
Yanyan Jia
Zuhua Yu
Zuhua Yu
Songbiao Chen
Songbiao Chen
Zhongyu Liu
Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolytica
Frontiers in Cellular and Infection Microbiology
Mannheimia haemolytica type A5
outer membrane vesicles
proteomics
inflammatory response
in vitro test
title Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolytica
title_full Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolytica
title_fullStr Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolytica
title_full_unstemmed Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolytica
title_short Proteomic analysis of outer membrane vesicles derived from the type A5 Strain of Mannheimia haemolytica
title_sort proteomic analysis of outer membrane vesicles derived from the type a5 strain of mannheimia haemolytica
topic Mannheimia haemolytica type A5
outer membrane vesicles
proteomics
inflammatory response
in vitro test
url https://www.frontiersin.org/articles/10.3389/fcimb.2025.1578027/full
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