Identification of 60 secreted proteins for Mycoplasma bovis with secretome assay

Zubair, Muhammad; Muhamed, Shafii Abdullahi; Khan, Farhan Anwar; Zhao, Gang; Menghwar, Harish; Faisal, Muhammad Salman; Zhang, Hui; Zhu, Xifang; Rasheed, Muhammad; Chen, Yingyu; Marawan, Marawan A.; Chen, Huanchun

doi:10.1016/j.micpath.2020.104135

Cited by 16 publications

(29 citation statements)

References 33 publications

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“…Interestingly, 13 previously reported secreted proteins were included in the 59 secreted proteins predicted in the present study with immunoinformatic tools and were classified as either classical or non-classical secreted proteins [ 21 ]. Among the final 22 immunogenic secreted proteins selected, MbovP579 [ 13 ], MbovNase [ 14 ], MbovP217, MbovP674, MbovP739 (confirmed by our laboratory but data not yet reported), MbovP274 and MbovP570 were confirmed in this study, lending support to the reliability of the immunogenic secreted proteins predicted.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2020

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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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Section: Discussionmentioning

confidence: 99%

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

et al. 2020

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“…The recombinant proteins were also able to react with M. bovis -positive cattle serum, however, with most seen by rMbov570. The increased expression of some cytokines to the recombinant proteins and their mutants, including immunoregulatory IL-6, IL-12 and IFN-γ, further confirmed the possible use of the two highly immunogenic secreted proteins, MbovP274 and MbovP570, as potential protective antigens and candidates for lipoprotein-based sub-unit vaccine known to induce an extended memory immune response [ 37 , 39 ].…”

Section: Mycoplasma Bovis Vaccinesmentioning

confidence: 94%

“…Novel M. bovis secreted proteins were identified using proteomic analysis of the M. bovis secretome [ 37 ]. Of the predicted total of 246 proteins, 60 secreted M. bovis proteins were finally selected.…”

Section: Mycoplasma Bovis Vaccinesmentioning

confidence: 99%

“…Additional analysis of these proteins showed that the functions they are involved in, such as multiple cellular activities, chromatic and DNA dynamics, the actual alignment and others, may be important in terms of M. bovis virulence and the disease pathogenesis. In the context of a possible vaccine candidate, one secreted protein, MbovP0581, known as ABC transporter ATP-binding protein, appeared to exhibit high immunogenic properties based on its reactivity with sera originated from cattle suffering from M. bovis pneumonia [ 37 ].…”

Section: Mycoplasma Bovis Vaccinesmentioning

confidence: 99%

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Recent Developments in Vaccines for Bovine Mycoplasmoses Caused by Mycoplasma bovis and Mycoplasma mycoides subsp. mycoides

2021

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Two of the most important diseases of cattle are caused by mycoplasmas. Mycoplasma bovis is a world-wide bovine pathogen that can cause pneumonia, mastitis and arthritis. It has now spread to most, if not all, cattle-rearing countries. Due to its increasing resistance to antimicrobial therapy, vaccination is the principal focus of the control of infection, but effective vaccines are currently lacking. Despite being eradicated from most parts of the world, Mycoplasma mycoides subsp. mycoides, the cause of contagious bovine pleuropneumonia (CBPP), continues to plague sub-Saharan Africa, affecting at least 25 countries. Numerous new experimental vaccines have been developed over the last 20 years to improve on protection afforded by the T1/44, a live vaccine in continuous use in Africa for over 60 years, but none so far have succeeded; indeed, many have exacerbated the disease. Tools for diagnosis and control are adequate for eradication but what is necessary are resources to improve vaccine coverage to levels last seen in the 1970s, when CBPP was restricted to a few countries in Africa. This paper summarizes the results of the main studies in the field of experimental mycoplasma vaccines, reviews data on commercially available bacterin vaccines and addresses issues relating to the search for new candidates for effective vaccines to reduce economic losses in the cattle industry caused by these two mycoplasmas.

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“…To exclude that, we extracted the greatest common divisor generated from these two approaches. Following this protocol, 178 overlapped proteins were identified, and 35 secreted proteins were previously identified with the 2-DE-based proteomic assay of M. bovis (27). Therefore, compared to the label-free proteomic technique, the conventional 2-DE-based proteomic assay missed many secreted proteins.…”

Section: A Total Of 178 Secreted Proteins Were Identified In M Bovis Virulent and Its Attenuated Strainsmentioning

confidence: 99%

Comparative Secretome Analyses of Mycoplasma bovis Virulent and Attenuated Strains Revealed MbovP0145 as a Promising Diagnostic Biomarker

Zhang

et al. 2021

Front. Vet. Sci.

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Mycoplasmas are successful pathogens both in humans as well as in animals. In cattle, Mycoplasma bovis (M. bovis) is known to be responsible for serious health complications, including pneumonia, mastitis, and arthritis. However, M. bovis pathogenesis remains unclear. Secreted proteins of M. bovis could influence infection and modify host defense signaling pathways after they enter their extracellular space in the host micro-environment. Therefore, this study was aimed to compare the secretomes of M. bovis HB0801 virulent (P1) and attenuated (P150) strains and identify potential pathogenesis-related secreted proteins and biomarkers. The cells of P1 and P150 strains were grown in pleuropneumonia-like organism medium to log phase and then transferred to phosphate-buffered saline for 2 h. Then, the supernatant was analyzed by using label-free quantitative proteomics, and 477 potential secreted proteins were identified. Combined with the bioinformatics prediction, we found that 178 proteins were commonly secreted by the P1 and P150 strains, and 49 of them were encoded by mycoplasmal core genes. Additionally, 79 proteins were found to have a different abundance between the P1 and P150 strains. Among these proteins, 34 were more abundant and uniquely expressed in P1, indicating a possible association with the virulence of M. bovis. Three differentially secreted proteins, MbovP0145, MbovP0725, and MbovP0174, as well as one equally secreted protein, MbovP0481, as positive control and one protein of inner membrane, MbovP0310, as negative control were, respectively, cloned, expressed, and evaluated for antigenicity, subcellular location, and the secretion nature with their mouse antisera by western blotting and colony immunoblotting assay. Among them, MbovP0145 was confirmed to be more secreted by P1 than P150 strain, highly reactive with the antisera from naturally infected and P1 experimentally infected cattle but not with the P150 vaccinated calves, indicating its potential as a diagnostic antigen. In conclusion, these findings may represent the most extensive compilation of potentially secreted proteins in mycoplasma species and the largest number of differentially secreted proteins between the virulent and attenuated M. bovis strains to date and provide new insights into M. bovis pathogenesis and diagnosis.

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Identification of 60 secreted proteins for Mycoplasma bovis with secretome assay

Cited by 16 publications

References 33 publications

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

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

Recent Developments in Vaccines for Bovine Mycoplasmoses Caused by Mycoplasma bovis and Mycoplasma mycoides subsp. mycoides

Comparative Secretome Analyses of Mycoplasma bovis Virulent and Attenuated Strains Revealed MbovP0145 as a Promising Diagnostic Biomarker

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