Development of a Multi-Epitope Vaccine for Mycoplasma hyopneumoniae and Evaluation of Its Immune Responses in Mice and Piglets

Abstract

Mycoplasma hyopneumoniae (Mhp), the primary pathogen causing Mycoplasma pneumonia of swine (MPS), brings massive economic losses worldwide. Genomic variability and post-translational protein modification can enhance the immune evasion of Mhp, which makes MPS prone to recurrent outbreaks on farms, even with vaccination or other treatments. The reverse vaccinology pipeline has been developed as an attractive potential method for vaccine development due to its high efficiency and applicability. In this study, a multi-epitope vaccine for Mhp was developed, and its immune responses were evaluated in mice and piglets. Genomic core proteins of Mhp were retrieved through pan-genome analysis, and four immunodominant antigens were screened by host homologous protein removal, membrane protein screening, and virulence factor identification. One immunodominant antigen, AAV27984.1 (membrane nuclease), was expressed by E. coli and named rMhp597. For epitope prioritization, 35 B-cell-derived epitopes were identified from the four immunodominant antigens, and 10 MHC-I and 6 MHC-II binding epitopes were further identified. The MHC-I/II binding epitopes were merged and combined to produce recombinant proteins MhpMEV and MhpMEVC6His, which were used for animal immunization and structural analysis, respectively. Immunization of mice and piglets demonstrated that MhpMEV could induce humoral and cellular immune responses. The mouse serum antibodies could detect all 11 synthetic epitopes, and the piglet antiserum suppressed the nuclease activity of rMhp597. Moreover, piglet serum antibodies could also detect cultured Mhp strain 168. In summary, this study provides immunoassay results for a multi-epitope vaccine derived from the reverse vaccinology pipeline, and offers an alternative vaccine for MPS.

Novel antigenic proteins of Mycoplasma synoviae as potential vaccine candidates

“…Nucleases of mycoplasmas are considered to play an important role in growth and pathogenesis 29 . In Mycoplasma hyopneumoniae, a nuclease (Mhp597) was screened as an immunodominant antigen, using reverse vaccinology pipeline genomic core proteins 30 . GrpE plays an important role in the survival of Mycoplasma during cellular stresses such as heat shock and hypoxia a well-known HSP70 cofactor 31,32 .…”

Conserved Domains in Variable Surface Lipoproteins A-G of Mycoplasma hyorhinis May Serve as Probable Multi-Epitope Candidate Vaccine: Computational Reverse Vaccinology Approach

“…Selected B and T cell epitopes stimulate innate and adaptive immunity, resulting in protective memory. Recently, the application of computational methods for vaccine design has been the preferred approach by researchers [ 35 , 36 , 37 , 38 ]. The proposed candidate vaccine, by using a computational reverse vaccinology approach, can be evaluated through animal immunization assay and has proved to be an effective strategy.…”

“…The proposed candidate vaccine, by using a computational reverse vaccinology approach, can be evaluated through animal immunization assay and has proved to be an effective strategy. A study conducted by Li and colleagues used the computational approach to propose the vaccine candidate and successfully evaluated its efficacy in mice models [ 37 ]. Implementing a computational reverse vaccinology approach and its success in in vivo experiments are supported by several studies [ 39 , 40 , 41 , 42 ].…”

Infection of Trichinella spiralis Affects the Reproductive Capacity of ICR/CD-1 Male Mice by Reducing the Urine Pheromone Contents and Sperm Quality

“…The serum separation was performed every 10 days and all serum samples were stored at −80 °C until use. The ELISA was performed with the previously published method [ 54 ]. In brief, the excretory-secretory antigen was diluted to 1 μg/mL with the antigen coating buffer (0.05 mol/L phosphate buffer solution, pH 9.6), and a 96-well microplate was coated with 100 μL of diluted antigen at 4 °C for twelve hours.…”