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In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification

Mycoplasma bovis is a major pathogen, responsible for bovine respiratory diseases worldwide. The present lack of effective control measures leaves cattle owners at considerable perpetual risk of M. bovis outbreaks. In this study, we identified M. bovis secreted immunogenic proteins in silico as pote...

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Autores principales: Shirani, Ihsanullah, Zhang, Hui, Zhao, Gang, Lu, Siyi, Marawan, Marawan A, Dawood, Ali, Chen, Yingyu, Chen, Xi, Chen, Jianguo, Hu, Changmin, Chen, Huanchun, Guo, Aizhen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559824/
https://www.ncbi.nlm.nih.gov/pubmed/32967149
http://dx.doi.org/10.3390/pathogens9090770
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author Shirani, Ihsanullah
Zhang, Hui
Zhao, Gang
Lu, Siyi
Marawan, Marawan A
Dawood, Ali
Chen, Yingyu
Chen, Xi
Chen, Jianguo
Hu, Changmin
Chen, Huanchun
Guo, Aizhen
author_facet Shirani, Ihsanullah
Zhang, Hui
Zhao, Gang
Lu, Siyi
Marawan, Marawan A
Dawood, Ali
Chen, Yingyu
Chen, Xi
Chen, Jianguo
Hu, Changmin
Chen, Huanchun
Guo, Aizhen
author_sort Shirani, Ihsanullah
collection PubMed
description Mycoplasma bovis is a major pathogen, responsible for bovine respiratory diseases worldwide. The present lack of effective control measures leaves cattle owners at considerable perpetual risk of M. bovis outbreaks. In this study, we identified M. bovis secreted immunogenic proteins in silico as potential candidates for novel diagnostic agents and vaccines. We used immunoinformatics to analyze 438 M. bovis proteins previously identified with a label-free proteomics analysis of virulent M. bovis HB0801 (P1) and its attenuated P150 strains. The subcellular localization of these proteins was preliminarily screened and 59 proteins were found to be secreted extracellular proteins. Twenty-seven of these proteins contained a large number of predictive T-cell epitopes presented by major histocompatibility complex (MHC) class I and II molecules. Twenty-two of these 27 proteins had a high number of conformational B-cell epitopes, predicted from the corresponding 3D structural templates, including one unique to P1, two unique to P150, and 19 common to both strains. Five proteins were selected for further validation, and two of these, MbovP274 and MbovP570, were successfully expressed and purified. Both were confirmed to be secretory and highly immunogenic proteins that induced a mouse antibody response, reacted with cattle serum positive for M. bovis infection, and significantly increased the production of interleukin 8 (IL-8), IL-12 and interferon γ (IFN-γ) during the secretion of these three cytokines by both M. bovis mutants of these genes. These results should be useful in the development of novel immunological agents against M. bovis infection.
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spelling pubmed-75598242020-10-29 In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification Shirani, Ihsanullah Zhang, Hui Zhao, Gang Lu, Siyi Marawan, Marawan A Dawood, Ali Chen, Yingyu Chen, Xi Chen, Jianguo Hu, Changmin Chen, Huanchun Guo, Aizhen Pathogens Article Mycoplasma bovis is a major pathogen, responsible for bovine respiratory diseases worldwide. The present lack of effective control measures leaves cattle owners at considerable perpetual risk of M. bovis outbreaks. In this study, we identified M. bovis secreted immunogenic proteins in silico as potential candidates for novel diagnostic agents and vaccines. We used immunoinformatics to analyze 438 M. bovis proteins previously identified with a label-free proteomics analysis of virulent M. bovis HB0801 (P1) and its attenuated P150 strains. The subcellular localization of these proteins was preliminarily screened and 59 proteins were found to be secreted extracellular proteins. Twenty-seven of these proteins contained a large number of predictive T-cell epitopes presented by major histocompatibility complex (MHC) class I and II molecules. Twenty-two of these 27 proteins had a high number of conformational B-cell epitopes, predicted from the corresponding 3D structural templates, including one unique to P1, two unique to P150, and 19 common to both strains. Five proteins were selected for further validation, and two of these, MbovP274 and MbovP570, were successfully expressed and purified. Both were confirmed to be secretory and highly immunogenic proteins that induced a mouse antibody response, reacted with cattle serum positive for M. bovis infection, and significantly increased the production of interleukin 8 (IL-8), IL-12 and interferon γ (IFN-γ) during the secretion of these three cytokines by both M. bovis mutants of these genes. These results should be useful in the development of novel immunological agents against M. bovis infection. MDPI 2020-09-21 /pmc/articles/PMC7559824/ /pubmed/32967149 http://dx.doi.org/10.3390/pathogens9090770 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Shirani, Ihsanullah
Zhang, Hui
Zhao, Gang
Lu, Siyi
Marawan, Marawan A
Dawood, Ali
Chen, Yingyu
Chen, Xi
Chen, Jianguo
Hu, Changmin
Chen, Huanchun
Guo, Aizhen
In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification
title In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification
title_full In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification
title_fullStr In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification
title_full_unstemmed In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification
title_short In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification
title_sort in silico identification of novel immunogenic secreted proteins of mycoplasma bovis from secretome data and experimental verification
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559824/
https://www.ncbi.nlm.nih.gov/pubmed/32967149
http://dx.doi.org/10.3390/pathogens9090770
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