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[Objective] Mycoplasmoides pneumoniae is a major pathogen responsible for community-acquired pneumonia worldwide, and the aggravating antibiotic resistance underscores the urgent need to develop an effective vaccine. [Methods] According to the critical pathogenic mechanisms of M. pneumoniae, 13 key adhesins and the community-acquired respiratory distress syndrome (CARDS) toxin were selected as candidate antigens. Reverse vaccinology and immunoinformatics approaches were adopted to predict and screen cytotoxic T lymphocyte (CTL) epitopes, helper T lymphocyte (HTL) epitopes, and linear B-cell epitopes. The screened epitopes were then connected via specific linkers. Two adjuvants, human beta-defensin 2 (HBD-2) and 50S ribosomal protein L7/L12, were introduced at the N-terminus to assemble the complete vaccine sequence. Furthermore, the physicochemical and biological properties of the constructed vaccine sequence were comprehensively analyzed. [Results] A total of 15 CTL epitopes, 10 HTL epitopes, and 14 linear B-cell epitopes were obtained after screening. All the epitopes and adjuvants were ligated by specific linkers, and a final multi-epitope vaccine sequence consisting of 746 amino acid residues was successfully constructed. The computational analysis indicated that the vaccine construct exhibited favorable physicochemical properties, antigenicity, solubility, and a safety profile. Molecular docking results showed that this construct could bind to Toll-like receptor 2 (TLR2) and TLR4 stably, which was further confirmed with the results of molecular dynamics simulations. The immune simulation and epitope coverage analysis indicated that the vaccine could elicit potent humoral and cellular immune responses and provide potential protection for up to 99.74% of the global population. [Conclusion] Through systematic computational design and evaluation, this study develops a multi-epitope mRNA candidate vaccine with promising application prospects, laying a theoretical foundation for subsequent experimental validation and development.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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