In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification
Open Access
- 20 September 2020
- Vol. 9 (9), 770
- https://doi.org/10.3390/pathogens9090770
Abstract
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.This publication has 36 references indexed in Scilit:
- Improved Method for Linear B-Cell Epitope Prediction Using Antigen’s Primary SequencePLOS ONE, 2013
- In Silico Identification of Epitopes in Mycobacterium avium subsp. paratuberculosis Proteins That Were Upregulated under Stress ConditionsClinical and Vaccine Immunology, 2012
- Comparative Geno-Plasticity Analysis of Mycoplasma bovis HB0801 (Chinese Isolate)PLOS ONE, 2012
- PSORTb 3.0: improved protein subcellular localization prediction with refined localization subcategories and predictive capabilities for all prokaryotesBioinformatics, 2010
- Prediction of Lipoprotein Signal Peptides in Gram-Positive Bacteria with a Hidden Markov ModelJournal of Proteome Research, 2008
- Characterization ofFrancisella tularensisOuter Membrane ProteinsJournal of Bacteriology, 2007
- Cytokine mRNA profiling of peripheral blood mononuclear cells from trypanotolerant and trypanosusceptible cattle infected withTrypanosoma congolensePhysiological Genomics, 2006
- Feature-based prediction of non-classical and leaderless protein secretion"Protein Engineering, Design and Selection", 2004
- Reliable prediction of T‐cell epitopes using neural networks with novel sequence representationsProtein Science, 2003
- Immortalization and characterization of bovine peritoneal macrophages transfected with SV40 plasmid DNAVeterinary Immunology and Immunopathology, 1995