Amino sequence of H5N1 AIV(GD/1/96) NP was parsed into octamers or nonamers according to the peptide-binding motifs of MHC class I molecules belonging to the B4, B12, B15, and B19 haplotypes. The 3D structures of those peptides and the MHC class I molecules were predicted using homology modelling method and then the binding affinity between each peptide and MHC class I molecule was analysed using molecular docking. Those peptides, which have correct binding conformation and high binding affinity, were predicted to be potential T cell epitopes in chicken. Predicted peptides 9 of 25 were synthesized and used to stimulate splenic lymphocytes collected from NP DNA vaccineimmunized SPF chickens. MHC class I molecule-restricted potential T-cell epitopes were confirmed by detecting of CD8 + T cell proliferation and interferon (IFN-c) expression.