Iranian Journal of Medical Sciences
Iranian Journal of Medical Sciences

Immunoinformatic and Structural Design of an Influenza A DNA Vaccine Based on Conserved Nucleoprotein Epitopes and Heat Shock Protein 70 Adjuvant

Articles InPress

Document Type : Original Article(s)

Authors

Sanaz Sadeh 1
Amir Ghaemi 2
Nazila Arbab Soleimani 1
Majid Moghbeli 1
Golnaz BahramAli 3

1 Department of Microbiology, Damghan Branch, Islamic Azad University, Damghan, Iran

2 Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran

3 Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran

10.30476/ijms.2026.109702.4506
Abstract
Background: Influenza A is a health concern, as it constantly mutates and changes antigens. The conventional vaccine is not very effective in providing immunity. We have designed and characterized a DNA multi-epitope vaccine based on conserved regions of the influenza A nucleoprotein (NP) and have assessed the immune response.
Methods: This is a bioinformatics study carried out in 2025 at the Pasteur Institute of Iran in Tehran. Epitopes were predicted using the immune epitope database and were screened for conservancy, antigenicity, toxicity, and allergenicity using the alignment-independent antigenicity prediction server (VaxiJen), toxicity prediction tool (ToxinPred), and allergenicity prediction tool (AllergenFP).
The vaccine contains a cell-penetrating peptide (CPP) derived from the hemagglutinin protein, a vascular endothelial growth factor 183 (VEGF-183) signal peptide, a tetanus toxoid (TT) helper peptide (830-844), NP peptides, and two repeats of heat shock protein 70 (Hsp70)(407-426). The peptides are arranged using flexible and rigid linkers. The structure was predicted using the self-optimized prediction method with alignment (SOPMA) and AlphaFold2. The structure was further analyzed using high-ambiguity-driven protein–protein docking (HADDOCK) with human leukocyte antigen (HLA) class I and II and toll-like receptor (TLR) 2 and 4, followed by a 200-ns molecular dynamics (MD) simulation.
Results: The docking study showed proper interaction of the selected peptides with HLA and murine BALB/c (H-2d) major histocompatibility complex (MHC) class I and II and the whole vaccine with innate immune system receptors. In silico immune simulation indicated cytokine release and T cell activation patterns.
Conclusion: The constructed multi-epitope DNA construct was predicted to be antigenic and non-toxic, with structural stability and immunogenic potential. These computational results support prioritizing this construct for experimental expression and murine preclinical validation (immunogenicity, safety, and protection).

Highlights

Sanaz Sadeh (Google Scholar)
Amir Ghaemi (Google Scholar)

Keywords

Immunoinformatics
Influenza virus strains
Molecular dynamics
Cell-penetrating peptide
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Iranian Journal of Medical Sciences

Articles InPress, Corrected Proof
Available Online from 19 October 2026


  • Received 17 November 2025
  • Revised 09 February 2026
  • Accepted 28 March 2026

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