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.
Highlights
Mycoplasma bovis (M. bovis) belongs to the genus Mycoplasma in the class Mollicutes and is the smallest self-replicating organism that lacks a cell wall [1]
Of the 438 proteins previously identified with label-free proteomic analysis, 59 were found to be extracellular secreted proteins. Among these 59 proteins, 27 contained a high number of predicted T-cell epitopes presented by Pathogens 2020, 9, x FOR PEER REVIEW
22 of these 27 proteins contained a high number of conformational B-cell epitopes predicted from the corresponding three-dimensional (3D) structural templates
Summary
Mycoplasma bovis (M. bovis) belongs to the genus Mycoplasma in the class Mollicutes and is the smallest self-replicating organism that lacks a cell wall [1]. It is a major infectious agent of beef and dairy cattle, causing several clinical conditions, including pneumonia, mastitis, arthritis, etc. M. bovis pneumonia has been reported widely in various parts of China, with dramatic economic losses [2,3]. Like other Mycoplasma species, M. bovis is intrinsically resistant to a wide variety of antimicrobial drugs because it lacks cell wall, a trait that confers tolerance of various effective chemotherapeutic agents commonly used in veterinary medicine. The development of safe and effective vaccines is the best way to control mycoplasma-related illnesses, save animal lives, and minimize the losses attributable to these diseases
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