Qiuxia Wang1,2,3, Zhihao Sun1,2,3, Jingzhi Li4, Tao Qin1,2,3, Hongwei Ma4, Sujuan Chen5,6,7,8, Daxin Peng9,10,11,12, Xiufan Liu1,2,13. 1. College of Veterinary Medicine, Yangzhou University, 48 East Wenhui Road, Yangzhou, Jiangsu, 225009, People's Republic of China. 2. Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225009, Jiangsu, People's Republic of China. 3. Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Yangzhou, 225009, Jiangsu, People's Republic of China. 4. Division of Nanobiomedicine, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, 215000, People's Republic of China. 5. College of Veterinary Medicine, Yangzhou University, 48 East Wenhui Road, Yangzhou, Jiangsu, 225009, People's Republic of China. Chensj@yzu.edu.cn. 6. Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225009, Jiangsu, People's Republic of China. Chensj@yzu.edu.cn. 7. Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Yangzhou, 225009, Jiangsu, People's Republic of China. Chensj@yzu.edu.cn. 8. Joint Laboratory Safety of International Cooperation of Agriculture & Agricultural-Products, Yangzhou, Jiangsu, 225009, People's Republic of China. Chensj@yzu.edu.cn. 9. College of Veterinary Medicine, Yangzhou University, 48 East Wenhui Road, Yangzhou, Jiangsu, 225009, People's Republic of China. pengdx@yzu.edu.cn. 10. Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225009, Jiangsu, People's Republic of China. pengdx@yzu.edu.cn. 11. Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Yangzhou, 225009, Jiangsu, People's Republic of China. pengdx@yzu.edu.cn. 12. Joint Laboratory Safety of International Cooperation of Agriculture & Agricultural-Products, Yangzhou, Jiangsu, 225009, People's Republic of China. pengdx@yzu.edu.cn. 13. Joint Laboratory Safety of International Cooperation of Agriculture & Agricultural-Products, Yangzhou, Jiangsu, 225009, People's Republic of China.
Abstract
BACKGROUND: Hemagglutinin is a major surface protein in influenza A virus (IAV), and HA2 is relative conserved among different IAVs. It will be meaningful to identify broad-spectrum epitopes based on the HA2 protein. RESULTS: Overlapping peptides of the HA2 protein of the H5N1 IAV A/Mallard/Huadong/S/2005 were synthesized and loaded on modified silica gel film to form a microarray, and antisera against different subtypes of IAVs were used to screen universal epitopes. The selected epitope was further confirmed by western blotting using anti-peptide immune serum and viruses rescued with amino acid substitution. The results showed that 485-FYHKCDNECME-495 of the H5 14th peptide in HA2 had broad-spectrum binding activity with antisera against H1, H3, H4, H5, H6, H7, H8, H9, and H10 subtype IAV. Substitution of amino acids (K or D) in rescued viruses resulted in decreased serum binding, indicating that they were critical residues for serum binding activity. In Immune Epitope Database, some epitopes containing 14-4 peptide were confirmed as MHC-II-restricted CD4 T cell epitope and had effects on releasing IL-2 or IFN. CONCLUSION: The identified epitope should be a novel universal target for detection and vaccine design and its ability to generate immune protection needs further exploration.
BACKGROUND: Hemagglutinin is a major surface protein in influenza A virus (IAV), and HA2 is relative conserved among different IAVs. It will be meaningful to identify broad-spectrum epitopes based on the HA2 protein. RESULTS: Overlapping peptides of the HA2 protein of the H5N1IAV A/Mallard/Huadong/S/2005 were synthesized and loaded on modified silica gel film to form a microarray, and antisera against different subtypes of IAVs were used to screen universal epitopes. The selected epitope was further confirmed by western blotting using anti-peptide immune serum and viruses rescued with amino acid substitution. The results showed that 485-FYHKCDNECME-495 of the H5 14th peptide in HA2 had broad-spectrum binding activity with antisera against H1, H3, H4, H5, H6, H7, H8, H9, and H10 subtype IAV. Substitution of amino acids (K or D) in rescued viruses resulted in decreased serum binding, indicating that they were critical residues for serum binding activity. In Immune Epitope Database, some epitopes containing 14-4 peptide were confirmed as MHC-II-restricted CD4 T cell epitope and had effects on releasing IL-2 or IFN. CONCLUSION: The identified epitope should be a novel universal target for detection and vaccine design and its ability to generate immune protection needs further exploration.
Authors: Suxiang Tong; Xueyong Zhu; Yan Li; Mang Shi; Jing Zhang; Melissa Bourgeois; Hua Yang; Xianfeng Chen; Sergio Recuenco; Jorge Gomez; Li-Mei Chen; Adam Johnson; Ying Tao; Cyrille Dreyfus; Wenli Yu; Ryan McBride; Paul J Carney; Amy T Gilbert; Jessie Chang; Zhu Guo; Charles T Davis; James C Paulson; James Stevens; Charles E Rupprecht; Edward C Holmes; Ian A Wilson; Ruben O Donis Journal: PLoS Pathog Date: 2013-10-10 Impact factor: 6.823
Authors: X Fan; A M Hashem; Z Chen; C Li; T Doyle; Y Zhang; Y Yi; A Farnsworth; K Xu; Z Li; R He; X Li; J Wang Journal: Mucosal Immunol Date: 2014-07-23 Impact factor: 7.313