|
|
Complete genome sequence analysis and mouse pathogenicity of canine H9N2 subtype influenza virus from Guangxi |
LI San-mu1, SUN Xiang-xiang2, KONG Zi-rong2, HE Qi-song3, MA Lin3, FENG Shu-ping3, YANG Ke-yan2, ZENG Yong-fang2, XIONG Yi3, YAN Jian-hua1 |
1. Medical College of Guangxi University,Nanning 530004,China;
2. College of Animal Science and Technology of Guangxi University, Nanning 530004, China;
3. Guangxi Center of Animal Disease Prevention and Control, Nanning 530001, China |
|
|
Abstract To analyze the whole genome sequence and pathogenicity of H9N2 subtype influenza virus carried by pet dogs in Guangxi, we amplified and sequenced the whole genome by RT-PCR. The amino acid homology and evolutionary tree were analyzed in DNA-Star and MEGA 6.0 software, and pathogenicity was studied through mouse infection experiments. Eight gene segments of 13 H9N2 strains were derived from 5 different lineages, which belonged to the new genotype. The amino acid sequence identity was 99.0% with respect to A/equine/Guangxi/3/2011 isolated from Guangxi. Six internal genes were closely related to the H7N9 strain from human influenza virus in 2013. The HA cleavage site of all strains was RSSR↓GLF, thus indicating low virulence. The amino acid sites HA226, NA119, NP375 and PB2 showed some changes, among which HA226 was changed to leucine L, which binds human SAα2, 6-Gal. In BALB/c mice infected with H9N2 virus isolated from healthy dogs, the virus did not replicate in vivo, but the strains Ca/GX/8 and Ca/GX/10 from cold dogs replicated in vivo and caused clear clinical symptoms but not death. The virulence of the virus was enhanced after passage in mice, a finding speculated to be caused by the changes in amino acids at positions PA623 and 611 of PB2. A total of 13 H9N2 subtype influenza virus strains obtained from dogs in Guangxi have avian origin, specifically bind human receptors and have a complex genetic evolution relationship, thus posing a great threat to mammals and humans. Therefore, prevention and control measures must urgently be strengthened.
|
Received: 22 October 2020
|
|
Fund:Supported by the National Natural Science Foundation of China (No. 31660713)and the Major Project of Natural Science Foundation of Guangxi (No. 2015GXNSFEA139002) |
Corresponding Authors:
Yan Jian-hua, Email:Jianhuayan@gxu.edu.cn
|
|
|
|
[1] Homme PJ, Easterday BC.Avian influenza virus infections. I. Characteristics of influenza A-turkey-Wisconsin-1966 virus[J]. Avian Dis, 1970,14(1):66-74. DOI:10.2307/1588557
[2] Shortridge KF, Stuart-Harris C.An influenza epicenter[J]. Lancet, 1982, 320(8302): 812-813. DOI: 10.1016/s0140-6736(82)92693-9
[3] Xu KM, Smith GJD, Bahl J, et al.The genesis and evolution of H9N2 influenza viruses in poultry from southern China, 2000 to 2005[J]. J Virol, 2007, 81(19):10389-10401. DOI: 10.1128/JVI.00979-07
[4] Panshin A, Golender N, Davidson I, et al.Variability of NS1 proteins among H9N2 avian influenza viruses isolated in Israel during 2000-2009[J]. Virus Genes, 2010, 41(3):396-405. DOI: 10.1007/s11262-010-0522-3
[5] Butt KM, Smith GJD, Chen H, et al.Human infection with an avian H9N2 influenza A virus in Hong Kong in 2003[J]. J Clin Microbiol, 2005, 43(11): 5760-5767. DOI: 10.1128/JCM.43.11.5760-5767.2005
[6] Song WJ, Qin K.Human-infecting influenza A (H9N2) virus: a forgotten potential pandemic strain?[J]. Zoonoses Public Health, 2020, 67(3):203-212.DOI: 10.1111/zph.12685
[7] Cheng VCC, Chan JFW, Wen X, et al.Infection of immunocompromised patients by avian H9N2 influenza A virus.[J]. J Infect, 2011, 62(5):394-399. DOI: 10.1016/j.jinf.2011.02.007
[8] Saito S, Takayama I, Nakauchi M, et al.Development and evaluation of a new real-time RT-PCR assay for detecting the latest H9N2 influenza viruses capable of causing human infection[J]. Microbiol Immunol, 2019, 63(1):21-31. DOI: 10.1111/1348-0421.12666
[9] 屠宇平,杨小平. 狗禽流感病毒(H5N1)实验感染和自然接触暴露[J]. 疾病监测, 2008, 23(2):130.
[10] Yamanaka T, Nemono M, Tsujimura K, et al.Interspecies transmission of equine influenza virus(H3N8) to dogs by close contact with experimentally infected horses[J]. Vet Microbiol, 2009, 139(3/4):351-355. DOI: 10.1016/j.vetmic.2009.06.015
[11] Song D, Lee C, Kang B,et al.Experiment infection of dogs with avian-origin canine influenza A virus(H3N2)[J]. Emerge Infect Dis, 2009, 15(1):56-58. DOI: 10.3201/eid1501.080755
[12] Chen Y, Zhong GX, Wang GJ, et al.Dogs are highly susceptible to H5N1 avian influenza virus[J]. Virology, 2010, 405(1):15-19. DOI: 10.1016/j.virol.2010.05.024
[13] Lin DG, Sun SS, Du LJ, et al.Natural and experimental infection of dogs with pandemic H1N1/2009 influenza virus[J]. J Gen Virol, 2012, 93(1):119-123. DOI: 10.1099/vir.0.037358-0
[14] Zhan GJ, Ling ZS, Zhu YL, et al.Genetic characterization of a novel influenza A virus H5N2 isolated from a dog in China[J]. Vet Microbiol, 2012, 155(2-4): 409-416.DOI: 10.1016/j.vetmic.2011.08.017
[15] Zhang K, Zhang ZW, Yu ZJ, et al.Domestic cats and dogs are susceptible to H9N2 avian influenza virus[J]. Virus Res, 2013, 175(1):52-57. DOI: 10.1016/j.virusres.2013.04.004
[16] Amirsalehy H, Ni LH, Mohammadi A.Can dogs carry the global pandemic candidate avian influenza virus H9N2[J]. Aust Vet J, 2012, 90(9):341-345. DOI: 10.1111/j.1751-0813.2012.00973.x
[17] Zhao FR, Liu CG, Yin X, et al.Serological report of pandemic (H1N1) 2009 infection among cats in Northeastern China in 2012-02 and 2013-03[J]. Virol J, 2014, 11(1):49. DOI: 10.1186/1743-422X-11-49
[18] Gamblin SJ, Haire LF, Russell RJ, et al.The structure and receptor binding properties of the 1918 influenza hemagglutinin[J]. Science, 2004, 303(5665):1838-1842. DOI: 10.1126/science.1093155
[19] Stieneke-Gröber A, Vey M, Angliker H, et al.Influenza virus hemagglutinin with multibasic cleavage site is activated by furin, a subtilisin-like endoprotease[J]. EMBO J,1992, 11(7): 2407-2414. DOI:10.1002/j.1460-2075.1992.tb05305.x
[20] 刘红旗, 程坚, 彭大新,等. 我国部分地区 H9 亚型禽流感病毒血凝素基因序列比较与遗传发生关系分析[J]. 微生物学报, 2002, 42(3): 288-297.DOI:10.13343/j.cnki.wsxb.2002.03.005
[21] Matrosovich M, Zhou N, Kawaoka Y, et al.The surface glycoproteins of H5 influenza viruses isolated from humans, chickens, and wild aquatic birds have distinguishable properties[J]. J Virol, 1999, 73(2): 1146-1155. DOI: 10.1128/JVI.73.2.1146-1155.1999
[22] Pinto LH, Holsinger LJ, Lamb RA.Influenza virus M2 protein has ion channel activity[J]. Cell, 1992, 69(3): 517-528. DOI:10.1016/0092-8674(92)90452-I
[23] 黄艳艳. 我国部分地区鸡源H9N2亚型AIV的系统进化和HA抗原分析以及AIV NA分型方法的研究[D]. 雅安:四川农业大学,2010.
[24] Gao RB, Cao B, Hu YW, et al.Human infection with a novel avian-origin influenza A (H7N9) virus[J]. N Engl J Med, 2013, 368(20):1888-1897. DOI: 10.1056/NEJMoa1304459
[25] Zhang QY, Shi JZ, Deng GH, et al.H7N9 influenza viruses are transmissible in ferrets by respiratory droplet[J]. Science, 2013, 341(6144):410-414. DOI: 10.1126/science.1240532
[26] Shi JZ, Deng GH, Liu PH, et al.Isolation and characterization of H7N9 viruses from live poultry markets-implication of the source of current H7N9 infection in humans[J]. Chin Sci Bull, 2013, 58(16):1857-1863. DOI: 10.1007/s11434-013-5873-4
[27] Cheung TKW, Poon LLM.Biology of influenza a virus[J]. Ann N Y Acad Sci, 2007,1102:1-25. DOI: 10.1196/annals.1408.001
[28] Hu Z, Zhang YR, Wang Z, et al.Mouse-adapted H9N2 avian influenza virus causes systemic infection in mice[J].Virol J, 2019, 16(1):135.DOI: 10.1186/s12985-019-1227-4 |
[1] |
LIU Yong-ning, MA Xin-ao, LYU Si-ying, LI Yuan, LI Xu-yong, SI Zhen-shu, GUO Jing, LI Yu-bao, LIU Cheng. ANXA2 knockout A549 cell line construction and effects on the replication of H1N1 and H9N2 influenza viruses[J]. Chinese Journal of Zoonoses, 2022, 38(4): 291-296. |
|
|
|
|