Abstract

Introduction: Avian influenza strains are responsible for approximately 650,000 deaths worldwide annually. H5N6 is a highly pathogenic virus that has already caused 30 acute respiratory distress syndrome-related deaths since 2014. The M2 proton channel is one of the virus? main antigens of importance. Aims: In this study we aimed to identify B- and T cell epitopes in the M2 protein of the H5N6 virus using in silico methodology. Methods: The M2 sequence in FASTA format was obtained from the Uniprot database. The tertiary structure of M2 was modeled using the PHYRE2 software. The antigenicity of M2 was calculated through the Vaxijen v.2.0 server. Linear as well as non-linear B-cell epitopes were found using the ElliPro tool of the IEDB database. MHC class I and II epitopes were identified using the Vaxitop tool of the Vaxign system. MHC I were further tested for their immunogenicity using the IEDB immunogenicity tool, while MHC II were tested for their ability to produce IFN-? using the IFNepitope server. The antigenicity of the predicted B- and T-cell epitopes was re-examined via Vaxijen and peptides that were identified as potential antigens were individually tested for the risk of allergenicity and toxicity as well as the possibility of inducing autoimmunity via the AllerTOP v. 2.0, Toxin Pred and Peptide Match tools respectively. Results: Our in-silico analysis resulted in the identification of 2 linear B-cell epitopes and 3 T-cell epitopes which fulfill prerequisites for vaccine design. Conclusions: Our findings suggest that M2 is a promising vaccine target against H5N6 yet, validation experiments are required to complement the immunoinformatic analysis.