Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (24): 5100-5107.doi: 10.3864/j.issn.0578-1752.2011.24.016

• VETERINARY SCIENCE • Previous Articles     Next Articles

Characteristics of Pathogenic and HA Antigenic Variation of H9N2 Subtybe Avian Influenza Viruses Isolated from 1998 to 2008 in China

 CHEN  Lu, LIU  Shou-Chuan, ZHAO  Jun, WANG  Chuan-Qing, WANG  Ze-Lin   

  1. 1.河南农业大学禽病研究所，郑州 450002
  • Received:2010-04-15 Online:2011-12-15 Published:2011-10-18

PDF

1202

Cited

2

Abstract: 【Objective】 The objective of this experiment is to investigate the characteristics of pathogenic and antigenic variation of H9N2 subtybe avian influenza viruses isolated from 1998 to 2008 in China. 【Method】 The EID50, ELD50, MDT, ICPI, IVPI and the duration of shedding virus from infected 8-week-old SPF chickens of different H9N2 avian influenza viruses isolates were determined. The HI and VN activity of monoclonal antibody 2A4 and F6 on different H9N2 avian influenza viruses isolates were assayed and the HA genes of different antigenic reactive isolates were sequenced and analyzed. 【Result】 The determined pathogenicity suggested that the virulence of different isolates were different, thereinto, 3#, 12#, and 14# isolates showed higher pathogenicity than the others and could cause the death of SPF chickens. The 8-week-old SPF chickens infected by 3# or 12# isolate shed virus earlier and last for a longer time. 3 # and 12 # isolates showed specific response properties to monoclonal antibody 2A4 and F6. Monoclonal antibody 2A4 and F6 could inhibit the hemagglutinin activity of 3 # and 12 # isolates, however it could not neutralize the virus infection on CEF cells. HA sequence analysis showed that there was a single amino acid substitution of Ser (S)-to- Asn(N) at position 145 in the HA protein of 3 # and 12 # isolates, which led to the loss of reactivity to the monoclonal antibody 2A4 and F6 and the occurrence of a new potential glycosylation site NGT. The change of reactivity to the monoclonal antibody 2A4 and F6 suggests that the site (S145) is one of HA protein epitope of the H9N2 subtype avian influenza A virus. The new occurring potential glycoprotein site NGT in the HA protein of 3 # and 12 # isolates may cause the enhancing pathogenicity. 【Conclusion】 The results show that H9N2 subtype avian influenza virus isolates have the tendency to evolve, resulting to the occurrence of mutants which have higher virulence and variable antigenicity. The higher virulent mutants may cause death of chickens and lead to more economic loss. The antigenic mutants may evade the immunity, which pose a new challenge to the immune prevention for the H9N2 avian influenza.

Key words: avian influenza virus, H9N2 subtype, HA glycoprotein, pathogenicity, antigenic variation

[1]Fouchier R A, Munster V, Wallensten A, Bestebroer T M, Herfst S, Smith D, Rimmelzwaan G F, Olsen B, Osterhaus A D. Characterization of a novel influenza A virus hemagglutinin subtype (H16) obtained from black-headed gulls. Journal of Virology, 2005, 79: 2814-2822.

[2]Homme P J, Easterday B C. Characteristics of influenza A-turkey- Wisconsin-1966 virus. Avian Disease, 1970, 14: 66-74.

[3]Zhang P, Tang Y, Liu X, Peng D, Liu W, Liu H, Lu S, Liu X. Characterization of H9N2 influenza viruses isolated from vaccinated flocks in an integrated broiler chicken operation in eastern China during a 5 year period (1998–2002) . Journal General Virology, 2008, 89, 3102-3112.

[4]Sun Y, Pu J, Jiang Z, Guan T, Xia Y, Xu Q, Liu L, Ma B, Tian F, Brown E G, Liu J. Genotypic evolution and antigenic drift of H9N2 influenza viruses in China from 1994 to 2008. Veterinary Microbiology，2010, 146(3-4): 215-225.

[5]Park K J, Kwon H I, Song M S, Pascua P N, Baek Y H, Lee J H, Jang H L, Lim J Y, Mo I P, Moon H J, Kim C J, Choi Y K. Rapid evolution of low-pathogenic H9N2 avian influenza viruses following poultry vaccination programmer. Journal General Virology, 2011, 92(1): 36-50.

[6]Rogers G N, Paulson J C, Daniels R S, Skehel J J, Wilson I A, Wiley D C. Single amino acid substitutions in influenza hemagglutinin change receptor binding specificity. Nature, 1983, 304: 76-78.

[7]Xu L, Bao L, Lv Q, Deng W, Ma Y, Li F, Zhan L, Zhu H, Ma C, Qin  C. A single-amino-acid substitution in the HA protein changes the replication and pathogenicity of the 2009 pandemic A (H1N1) influenza viruses in vitro and in vivo. Virology Journal, 2010, 7: 32-35.

[8]Xu Q, Wang W, Cheng X, Zengel J, Jin H. Influenza H1N1 A/Solomon Island/3/06 virus receptor binding specificity correlates with virus pathogenicity, antigenicity, and immunogenicity in ferrets. Journal of Virology, 2010, 84: 4936-4945.

[9]Ping J, Li C, Deng G, Jiang Y, Tian G, Zhang S, Bu Z, Chen H. Single-amino-acid mutation in the HA alters the recognition of H9N2 influenza virus by a monoclonal antibody. Biochemical and Biophysical Research Communications, 2008, 371(1): 168-171.

[10]中华人民共和国农业部. 中华人民共和国兽用生物制品质量规程（2001年版）. 北京: 中国农业出版社, 2001.

Agricultural Ministry of the People's Republic of China. Quality regulations of Veterinary Biologics of the People's Republic of China (2001 edition). Beijing: China Agricultural Press, 2001.( in Chinese)

[11]殷 震, 刘景华. 动物病毒学(第二版). 北京: 中国农业出版社, 1997.

Yin Z, Liu JH. Animal Virology (2nd edition). Beijing: China Agricultural Press, 1997. ( in Chinese)

[12]Wiley D C, Wilson I A, Skehel J J. Structural identification of the antibody-binding sites of Hong Kong influenza hemagglutinin and their involvement in antigenic variation. Nature, 1981, 289: 373-378.

[13]Caton A J, Brownlee G G, Yewdell J W, Gerhard W. The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin. Cell, 1982, 31: 417-427.

[14]Tsuchiya E, Sugawara K, Hongo S, Matsuzaki Y, Muraki Y, Li ZN, Nakamura K. Antigenic structure of the hemagglutinin of human influenza A/H2N2 virus. Journal of General Virology, 2001, 82: 2475-2484.

[15]Philpott M, Hioe C, Sheerar M, Hinshaw VS. Hemagglutinin mutations related to attenuation altered cell tropism of a virulent avian influenza A virus. Journal of Virology, 1990, 64: 2941-2947.

[16]Okamatsu M, Sakoda Y, Kishida N, Isoda N, Kida H. Antigenic structure of the hemagglutinin of H9N2 influenza viruses. Archives of Virology, 2008, 153(12): 2189-2195.

[17]Kaverin N V, Rudneva I A, Ilyushina N A, Lipatov A S, Krauss S, Webster R G. Structural differences among hemagglutinins of influenza a virus subtypes are reflected in their antigenic architecture: Analysis of H9 escape mutants. Journal of Virology, 2004, 78(1): 240-249.

[18]Ferreira H L, Lambrecht B, van Borm S, Torrieri-Dramard L, Klatzmann D, Bellier B, van den Berg T. Identification of a dominant epitope in the hemagglutinin of an Asian highly pathogenic avian influenza H5N1 clade 1 virus by selection of escape mutants. Avian Disease, 2010, 54(1): 565-571.

[19]Wei J, Yan B, Chen Z, Li T, Deng F, Wang H, Hu Z. Production and characterization of monoclonal antibodies against the hemagglutinin of H5N1 and antigenic investigation of avian influenza H5N1 viruses isolated from China. Canadian Journal of Microbiology, 2011, 57(1): 42-48.

[20]Tsuchiya E, Sugawara K, Hongo S, Matsuzaki Y, Muraki Y, Li Z N, Nakamura K. Effect of addition of new oligosaccharide chains to the globular head of influenza A/H2N2 virus hemagglutinin on the intracellular transport and biological activities of the molecule. Journal of General Virology, 2002, 83: 1137-1146.

[21]Skehel J J, Stevens D J, Daniels R S, Douglas A R, Knossow M, Wilson I A, Wiley D C. A carbohydrate side chain on hemagglutinins of Hong Kong influenza viruses inhibits recognition by a monoclonal antibody. Proceedings of the National Academy of Sciences of the United States of American, 1984, 81: 1779-1783.

[22]Schulze I T. Effects of glycosylation on the properties and functions of influenza virus hemagglutinin. The Journal of Infectious  Diseases, 1997, 176: 24-28.

[23]Kaverin N V, Rudneva I A, Ilyushina N A, Varich N L, Lipatov A S, Smirnov Y A, Govorkova E A, Gitelman A K, Lvov D K, Webster R G. Structure of antigenic sites on the hemagglutinin molecule of H5 influenza virus and phenotypic variation of escape mutants. Journal of General Virology, 2002, 83: 2497-2505.

[24]刘红旗, 张评浒, 刘秀梵, 刘文博, 贾立军. 封闭式饲养鸡场H9N2亚型禽流感病毒HA基因在5年内的遗传变异. 微生物学报, 2003, 43 (6): 706-711.

Liu H Q, Zhang P H, Liu X F, Liu W B, Jia L J. Genetic M utations of heam agglutinin genes of H9N2 subtype influenza a viruses in the field in a five- year period. Acta Microbiologica Sinica, 2003, 43 (6): 706-711.( in Chinese)

[25]娄本红, 朱秀同, 孙贝贝, 崔治中. 抗体选择压作用下H9N2亚型禽流感病毒HA基因的变异. 微生物学报, 2009, 49 (7) : 955-959.

Lou B H, Zhu X T, Sun B B, Cui Z Z. Mutations of the hemagglutinin gene of H9N2 subtype avian influenza viruses under selective pressure of antibody. Acta Microbiologica Sinica, 2009, 49(7): 955-959. ( in Chinese) 

[26]刘红旗, 黄 勇, 程 坚, 彭大新, 贾立军, 张如宽, 刘秀梵. 在疫苗免疫选择压力下H9N2亚型禽流行性感冒病毒HA基因的遗传变异. 病毒学报, 2002,18(2): 150-154.

Liu H Q, Huang Y, Cheng J, Peng D X, Jia L J Zhang R K, Liu X F. Genetic mutations of the hemagglutinin gene of H9N2 subtype avian influenza viruses under the selective pressure of vaccination. Chinese Journal of Virology, 2002, 18(2): 150-154.( in Chinese)
[1] MIAO JiaHao, CUI PengFei, YAN Cheng, WANG CongCong, WANG Yan, CHEN Yuan, CHEN Peng, SHI JianZhong, DENG GuoHua, CHEN HuaLan. Genetic and Biological Characterization of Two H8N4 Subtype Avian Influenza Viruses Isolated from Duck [J]. Scientia Agricultura Sinica, 2026, 59(7): 1576-1586.
[2] DONG Yu, WU Qian, FENG Xuan, ZHENG YinYing, CUI BaiMing. A Novel Plasmid pEA60 of Erwinia amylovora Enhances the Pathogenicity of Strains by Regulating the Synthesis of Virulence Factors [J]. Scientia Agricultura Sinica, 2026, 59(5): 996-1007.
[3] CONG QiQi, ZHANG JingYi, MENG XiangLong, DAI PengBo, LI Bo, HU TongLe, WANG ShuTong, CAO KeQiang, WANG YaNan. Identification of Hypovirus in Apple Ring Rot Fungus Botryosphaeria dothidea and Detection of Virus-Carrying Status in China [J]. Scientia Agricultura Sinica, 2025, 58(3): 478-492.
[4] TONG ZhaoYang, LIU WenHua, ZHANG GuoXin, DONG ChunYan, ZHANG YanXia, XU XiaoWei, HE Dong, LIU HeChun, LI Yang, WANG FengTao, FENG Jing, YAO XiaoBo, LIU MeiJin, LIN RuiMing. The Relationship Between Occurrence of Hulless Barley Ear Rot and Population Migration of Grass Mite (Siteroptes spp.) [J]. Scientia Agricultura Sinica, 2025, 58(3): 493-506.
[5] YANG WenJuan, GAO JiaCheng, WANG YanTing, LI Yan, GUO Ming, WANG JunCheng, MENG YaXiong, WANG HuaJun, SI ErJing. Function of Effector Pg00778 Regulation on the Pathogenicity of Pyrenophora graminea to Barley [J]. Scientia Agricultura Sinica, 2025, 58(15): 3020-3035.
[6] CHEN Yuan, CUI PengFei, SHI JianZhong, ZHANG YuanCheng, YU QingQing, YAN Cheng, ZHANG YaPing, WANG CongCong, ZHANG Jie, WANG Yan, DENG GuoHua, CHEN HuaLan. Biological Characteristics of H6N1 Subtype Avian Influenza Virus from 2019 to 2022 in China [J]. Scientia Agricultura Sinica, 2024, 57(9): 1820-1832.
[7] DONG ZaiFang, DING TengTeng, SHAN YiXuan, LI HongLian, CHEN LinLin, XING XiaoPing. Autophagy-Related Gene FpAtg3 Involves in Growth and Pathogenicity of Fusarium pseudograminearum [J]. Scientia Agricultura Sinica, 2024, 57(6): 1080-1090.
[8] ZHANG AiHong, YANG Fei, ZHAO YuanYe, ZHAO YiHan, DI DianPing, MIAO HongQin. Pathogenicity and Epidemic Risk of Barley Yellow Striate Mosaic Virus [J]. Scientia Agricultura Sinica, 2024, 57(23): 4686-4697.
[9] WANG Li, LI Kun, HUANG ShuLun, LI FengJuan, WANG Sheng, LU ZengJun, LIU ZaiXin. Antigenic Variation of Foot-and-Mouth Disease Virus Serotype O in Southeast Asia Topotype [J]. Scientia Agricultura Sinica, 2024, 57(15): 3093-3104.
[10] WANG Yuan, DU MengDan, LI ZhengGang, SHE XiaoMan, YU Lin, LAN GuoBing, DING ShanWen, HE ZiFu, TANG YaFei. Identification of Pathogen Causing Tomato White Tip and Curl Leaf Disease and Its Pathogenicity in Guangdong Province [J]. Scientia Agricultura Sinica, 2024, 57(12): 2350-2363.
[11] ZHANG Jian, ZHAO BinSen, FENG Hao, HUANG LiLi. Function and Mechanism Analysis of Vm-milRN7 Regulating the Pathogenicity of Valsa mali [J]. Scientia Agricultura Sinica, 2024, 57(10): 1930-1942.
[12] GAO XiaoXiao, TU LiQin, YANG Liu, LIU YaNan, GAO DanNa, SUN Feng, LI Shuo, ZHANG SongBai, JI YingHua. Construction of an Infectious Clone of Tobacco Mild Green Mosaic Virus Isolate Infecting Pepper from Jiangsu Based on Genomic Clone [J]. Scientia Agricultura Sinica, 2023, 56(8): 1494-1502.
[13] GONG AnDong, LEI YinYu, WU NanNan, LIU JingRong, SONG MengGe, ZHANG YiMei, YANG Guang, YANG Peng. The Effect of 3-Oxyacyl ACP Reductase Gene FgOAR1 on the Growth, Development and Pathogenicity of Fusarium graminearum [J]. Scientia Agricultura Sinica, 2023, 56(24): 4854-4865.
[14] LI HuiXin, SONG WenPing, HAN ZongXi, LIU ShengWang. Isolation and Pathogenicity of Fowl Adenovirus Serotype 8a Strain [J]. Scientia Agricultura Sinica, 2023, 56(16): 3226-3236.
[15] MA Qi, HE XinWen, WANG Yan, LIU YanJing, PAN ShuXin, HOU YuJie, SHI JianZhong, DENG GuoHua, BAO HongMei, LIU JingLi, GUO XingFu, MAO ShengGang, HU JingLei, LU Tong, YANG Fan, TIAN GuoBin, ZENG XianYing, CHEN HuaLan. The Protective Efficacy of Commercial Vaccines Against H9N2 Avian Influenza Virus of Branch h9.4.2.5 Isolated in China [J]. Scientia Agricultura Sinica, 2023, 56(15): 3040-3050.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd