Chimeric protein consisting of 3M2e and HSP as a universal influenza vaccine candidate: from in silico analysis to preliminary evaluation
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The 23-amino acid ectodomain of influenza virus M2 protein (M2e) is highly conserved among human influenza virus variants and represents an attractive target for developing a universal vaccine. Although this peptide has limited potency and low immunogenicity, the degree of M2e density has been shown to be a critical factor influencing the magnitude of epitope-specific responses. The aim of this study was to design a chimer protein consisting of three tandem repeats of M2e peptide sequence fused to the Leishmania major HSP70 gene and evaluate its characteristics and immunogenicity. The structure of the deduced protein and its stability, aliphatic index, biocomputed half-life and the anticipated immunogenicity were analyzed by bioinformatics software. The oligonucleotides encoding 3M2e and chimer 3M2e-HSP70 were expressed in Escherichia coli and affinity purified. The immunogenicity of the purified recombinant proteins was preliminary examined in mouse model. It was predicted that fusion of HSP70 to the C-terminal of 3M2e peptide led to increased stability, hydropathicity, continuous B cell epitopes and antigenic propensity score of chimer protein. Also, the predominant 3M2e epitopes were not hidden in the chimer protein. The initial in vivo experiment showed that 3M2e-HSP chimer protein stimulates specific immune responses. In conclusion, the results of the current study suggest that 3M2e-HSP chimer protein would be an effective universal subunit vaccine candidate against influenza infection.
KeywordsFusion protein Influenza virus M2e HSP70 in silico Universal vaccine
We would like to thank Dr. Masoumeh Tavassoti Kheiri and Dr. Reza Ahangari Cohan for kindly editing the manuscript.
Fatemeh Fotouhi, Behrokh Farahmand and Najmeh Taheri designed the construct, performed the analysis and wrote the manuscript. Hadiseh Shokouhi evaluated immune responses. Fatemeh Fotouhi supervised this work. Hoorieh Soleimanjahi reviewed and approved the final manuscript.
This work was funded by Grant No. 759 from Pasteur Institute of IRAN.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving in animals participant
Pasteur Institute of Iran, Ethic code: IR.PII.REC.1394.39.
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