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Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2
Haemophilus parasuis is a commensal organism of the upper respiratory tract of pigs, but virulent strains can cause Glässer’s disease, resulting in significant economic losses to the swine industry. OmpP2 is an outer membrane protein of this organism that shows considerable heterogeneity between vir...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10189045/ https://www.ncbi.nlm.nih.gov/pubmed/37207184 http://dx.doi.org/10.3389/fcimb.2023.1192651 |
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author | Wu, Jingbo Nan, Wenjin Peng, Guoliang Hu, Honghui Xu, Chongbo Huang, Jianqiang Xiao, Zhengzhong |
author_facet | Wu, Jingbo Nan, Wenjin Peng, Guoliang Hu, Honghui Xu, Chongbo Huang, Jianqiang Xiao, Zhengzhong |
author_sort | Wu, Jingbo |
collection | PubMed |
description | Haemophilus parasuis is a commensal organism of the upper respiratory tract of pigs, but virulent strains can cause Glässer’s disease, resulting in significant economic losses to the swine industry. OmpP2 is an outer membrane protein of this organism that shows considerable heterogeneity between virulent and non-virulent strains, with classification into genotypes I and II. It also acts as a dominant antigen and is involved in the inflammatory response. In this study, 32 monoclonal antibodies (mAbs) against recombinant OmpP2 (rOmpP2) of different genotypes were tested for reactivity to a panel of OmpP2 peptides. Nine linear B cell epitopes were screened, including five common genotype epitopes (Pt1a, Pt7/Pt7a, Pt9a, Pt17, and Pt19/Pt19a) and two groups of genotype-specific epitopes (Pt5 and Pt5-II, Pt11/Pt11a, and Pt11a-II). In addition, we used positive sera from mice and pigs to screen for five linear B-cell epitopes (Pt4, Pt14, Pt15, Pt21, and Pt22). After porcine alveolar macrophages (PAMs) were stimulated with overlapping OmpP2 peptides, we found that the epitope peptides Pt1 and Pt9, and the loop peptide Pt20 which was adjacent epitopes could all significantly upregulated the mRNA expression levels of IL-1α, IL-1β, IL-6, IL-8, and TNF-α. Additionally, we identified epitope peptides Pt7, Pt11/Pt11a, Pt17, Pt19, and Pt21 and loop peptides Pt13 and Pt18 which adjacent epitopes could also upregulate the mRNA expression levels of most proinflammatory cytokines. This suggested that these peptides may be the virulence-related sites of the OmpP2 protein, with proinflammatory activity. Further study revealed differences in the mRNA expression levels of proinflammatory cytokines, including IL-1β and IL-6, between genotype-specific epitopes, which may be responsible for pathogenic differences between different genotype strains. Here, we profiled a linear B-cell epitope map of the OmpP2 protein and preliminarily analyzed the proinflammatory activities and effects of these epitopes on bacterial virulence, providing a reliable theoretical basis for establishing a method to distinguish strain pathogenicity and to screen candidate peptides for subunit vaccines. |
format | Online Article Text |
id | pubmed-10189045 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-101890452023-05-18 Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2 Wu, Jingbo Nan, Wenjin Peng, Guoliang Hu, Honghui Xu, Chongbo Huang, Jianqiang Xiao, Zhengzhong Front Cell Infect Microbiol Cellular and Infection Microbiology Haemophilus parasuis is a commensal organism of the upper respiratory tract of pigs, but virulent strains can cause Glässer’s disease, resulting in significant economic losses to the swine industry. OmpP2 is an outer membrane protein of this organism that shows considerable heterogeneity between virulent and non-virulent strains, with classification into genotypes I and II. It also acts as a dominant antigen and is involved in the inflammatory response. In this study, 32 monoclonal antibodies (mAbs) against recombinant OmpP2 (rOmpP2) of different genotypes were tested for reactivity to a panel of OmpP2 peptides. Nine linear B cell epitopes were screened, including five common genotype epitopes (Pt1a, Pt7/Pt7a, Pt9a, Pt17, and Pt19/Pt19a) and two groups of genotype-specific epitopes (Pt5 and Pt5-II, Pt11/Pt11a, and Pt11a-II). In addition, we used positive sera from mice and pigs to screen for five linear B-cell epitopes (Pt4, Pt14, Pt15, Pt21, and Pt22). After porcine alveolar macrophages (PAMs) were stimulated with overlapping OmpP2 peptides, we found that the epitope peptides Pt1 and Pt9, and the loop peptide Pt20 which was adjacent epitopes could all significantly upregulated the mRNA expression levels of IL-1α, IL-1β, IL-6, IL-8, and TNF-α. Additionally, we identified epitope peptides Pt7, Pt11/Pt11a, Pt17, Pt19, and Pt21 and loop peptides Pt13 and Pt18 which adjacent epitopes could also upregulate the mRNA expression levels of most proinflammatory cytokines. This suggested that these peptides may be the virulence-related sites of the OmpP2 protein, with proinflammatory activity. Further study revealed differences in the mRNA expression levels of proinflammatory cytokines, including IL-1β and IL-6, between genotype-specific epitopes, which may be responsible for pathogenic differences between different genotype strains. Here, we profiled a linear B-cell epitope map of the OmpP2 protein and preliminarily analyzed the proinflammatory activities and effects of these epitopes on bacterial virulence, providing a reliable theoretical basis for establishing a method to distinguish strain pathogenicity and to screen candidate peptides for subunit vaccines. Frontiers Media S.A. 2023-05-03 /pmc/articles/PMC10189045/ /pubmed/37207184 http://dx.doi.org/10.3389/fcimb.2023.1192651 Text en Copyright © 2023 Wu, Nan, Peng, Hu, Xu, Huang and Xiao https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular and Infection Microbiology Wu, Jingbo Nan, Wenjin Peng, Guoliang Hu, Honghui Xu, Chongbo Huang, Jianqiang Xiao, Zhengzhong Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2 |
title | Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2 |
title_full | Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2 |
title_fullStr | Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2 |
title_full_unstemmed | Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2 |
title_short | Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2 |
title_sort | screening of linear b-cell epitopes and its proinflammatory activities of haemophilus parasuis outer membrane protein p2 |
topic | Cellular and Infection Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10189045/ https://www.ncbi.nlm.nih.gov/pubmed/37207184 http://dx.doi.org/10.3389/fcimb.2023.1192651 |
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