Leveraging the power of computational immunology to develop a novel couplet mRNA vaccine concatenating conserved epitopes of monkeypox virus antigens
DOI:
https://doi.org/10.24193/subbbiol.2026.1.05Keywords:
epitope, immunoinformatics, monkeypox, Mpox, mRNA vaccinesAbstract
The monkeypox (Mpox) disease was declared as a Public Health Emergency of International Concern (PHEIC) in 2022 and in 2024. Currently, there are no Mpox-specific vaccines and antiviral drugs for the treatment of Mpox. This study developed a novel couplet mRNA vaccine through immunoinformatics tools and databases. Highly conserved sequences of nine Mpox antigens were identified and utilized for T-cell and B-cell epitope mapping. Using data obtained from experimentally validated epitopes, the potential immunogenicity of predicted T-cell epitopes was evaluated. Peptides that may cause cross-reactivity, toxicity, and allergic reactions were excluded. The population coverage of candidate epitopes was also estimated. Epitopes were arranged and adjoined to form the mRNA constructs, aiming to direct its processing either to exogenous or endogenous pathways, while maximizing cleavage scores to produce the desired peptides. Conservancy analysis revealed that the nine Mpox target antigens A29L, A35R, A42R, B6R, E8L, F13L, H3L, L1R, and M1R are highly conserved. After the exclusion of potentially harmful and non-immunogenic peptides, 559 CD4+ and 104 CD8+ T-cell epitopes remained. The couplet mRNA constructs encode potentially immunogenic T-cell epitopes and accessible B-cell epitopes of multiple Mpox antigens that can be expressed and processed to generate the desired peptide sequences in humans. Estimated population coverage for mTcPox and mBThPox constructs are 77.67% and 81.81%, respectively. This study developed a novel couplet mRNA Mpox vaccine design containing highly conserved, immunogenic epitopes, with acceptable safety profile. Further in vitro, animal, and clinical studies are anticipated for human applications.
Article history: Received 9 August 2025; Revised 2 December 2025;
Accepted 16 March 2026; Available online 25 June 2026.
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