title: Exploration of the BF2*15 major histocompatibility complex class I binding motif and identification of cytotoxic T lymphocyte epitopes from the H5N1 influenza virus nucleoprotein in chickens.
abstract: The binding motif of BF2*15 major histocompatibility complex (MHC) class I was explored by analyzing the interaction between an infectious bronchitis virus octapeptide and BF2*15, and the cytotoxic T lymphocyte (CTL) epitope from the nucleoprotein (NP) of H5N1 virus was identified using experimental methods. Computational methods, including homology modeling, molecular dynamics simulation, and molecular docking analysis, were used. The recombinant plasmid pCAGGS-NP was constructed, and NP expression was confirmed by indirect immunofluorescence and Western blot in transfected 293T cells. Antibodies against NP in pCAGGS-NP-inoculated specific-pathogen-free chickens were detected by enzyme-linked immunosorbent assay (ELISA). Interferon γ (IFN-γ) mRNA was quantified, and IFN-γ production was evaluated using quantitative reverse transcription PCR and capture ELISA, respectively. CD8(+) T-lymphocyte proliferation was detected using flow cytometric analysis. The BF2*15 MHC class I binding motif "x-Arg/Lys-x-x-x-Arg/Lys" was explored. Quantification of chicken IFN-γ mRNA, evaluation of IFN-γ production, and measurement of CD8(+) T-lymphocyte proliferation confirmed that the peptide NP67-74 of H5N1 was the BF2*15 MHC-class-I-restricted CTL epitope.
authors: School of Social Development and Public Policy, China Institute of Health, Beijing Normal University, 19 Xinjiekou Wai St., Beijing, 100875, People's Republic of China.
authors: Weijun
authors: W
authors: Zhang
authors: Shandong Key Lab of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, Shandong, People's Republic of China.
authors: Qinghua
authors: Q
authors: Huang
authors: Weifang Engineering Vocational College, Qingzhou, 262500, Shandong, People's Republic of China.
authors: Mei
authors: M
authors: Lu
authors: College of Animal Science, Shandong Agricultural University, Tai'an, 271018, Shandong, People's Republic of China.
authors: Fengzhu
authors: F
authors: Zhu
authors: Shandong Key Lab of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, Shandong, People's Republic of China.
authors: Yan-Yan
authors: YY
authors: Huang
authors: Shandong Key Lab of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, Shandong, People's Republic of China.
authors: Shao-Hua
authors: SH
authors: Yang
authors: College of Animal Science, Shandong Agricultural University, Tai'an, 271018, Shandong, People's Republic of China.
authors: Zhengjie
authors: Z
authors: Kong
authors: Shandong Key Lab of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, Shandong, People's Republic of China.
authors: Xiu-Mei
authors: XM
authors: Zhang
authors: Shandong Key Lab of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, Shandong, People's Republic of China. xcttaian2002@163.com.
authors: Chuan-Tian
authors: CT
authors: Xu
mesh_terms: D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002645:Chickens; D018984:Epitopes, T-Lymphocyte / Q000378:metabolism*; D015235:HLA-B Antigens / Q000378:metabolism*; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D062105:Molecular Docking Simulation; D019590:Nucleocapsid Proteins; D011485:Protein Binding; D016601:RNA-Binding Proteins / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014758:Viral Core Proteins / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D015235:HLA-B Antigens; C048259:HLA-Bf antigen; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins