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

被引:11
作者
Li, Gaojian [1 ]
Shu, Jinqi [1 ]
Jin, Jing [1 ]
Shu, Jianhong [1 ,2 ]
Feng, Huapeng [1 ]
Chen, Jian [1 ]
He, Yulong [1 ,2 ]
机构
[1] Zhejiang Sci Tech Univ, Dept Biopharm, Coll Life Sci & Med, Hangzhou 310018, Peoples R China
[2] Shaoxing Biomed Res Inst, Zhejiang Sci Tech Univ, Shaoxing 312000, Peoples R China
关键词
Mycoplasma hyopneumoniae; reverse vaccinology; multi-epitope vaccine design; immunization of mice and piglets; immune response evaluation; B-CELL EPITOPES; PROTEIN-STRUCTURE; PREDICTION; SURFACE; ADHESIN; SERVER; IMMUNIZATION; LIPOPROTEIN; ADJUVANTS; ANTIGENS;
D O I
10.3390/ijms23147899
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页数:22
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