欧亚类禽型H1N1猪流感病毒HA蛋白的表达及免疫原性评估
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摘要
【目的】表达欧亚类禽型(eurasian avian-like,EA)H1N1猪流感病毒(SIV)的血凝素(HA)蛋白,并对其免疫原性进行测定。【方法】利用RT-PCR技术扩增病毒A/swine/Zhejiang/245/2013(H1N1)(ZJ245)的HA基因,将其克隆至真核表达载体pCAGGS中,获得重组质粒pCAGGS-HA(ZJ245),将其转染293T细胞,采用间接免疫荧光(IFA)检测和Western blot检测HA蛋白在体外的表达;将重组质粒pCAGGS-HA(ZJ245)经肌肉注射途径、以100μg/只剂量免疫16只6周龄BALB/c小鼠(I组和II组),间隔3周后进行加强免疫(二免);同等数量的小鼠以相同方式注射100μL无菌PBS作为非免疫对照组(III组和IV组)。首免和二免每周采血,分别采用血凝抑制(HI)试验和病毒中和(VN)试验两种方法测定不同类型的血清抗体效价;二免2周后,其中两组小鼠(I组和III组)用50μL(106.0EID50)的ZJ245病毒经滴鼻感染途径进行攻毒,另外两组(II组和IV组)用A/swine/Heilongjiang/44/2009(H1N1)(HLJ44)病毒进行攻毒。攻毒后14 d内每天观察小鼠的临床症状、统计发病与死亡情况,且每天称量小鼠体重,在体重下降比率超过25%时判定为小鼠死亡。攻毒后第3天,每组随机剖杀3只小鼠,分别采集脑、鼻甲、肺、脾和肾等脏器,匀浆处理后通过接种10日龄的非免疫鸡胚测定脏器的病毒含量。通过小鼠体重变化以及脏器滴定的病毒含量评估重组质粒pCAGGS-HA(ZJ245)的免疫保护效果。【结果】经酶切鉴定和测序验证表明ZJ245的HA基因的已正确克隆至真核表达质粒p CAGGS中获得重组质粒pCAGGS-HA(ZJ245),经体外转染293T细胞后,IFA和Western blot证实了病毒的HA蛋白能够正确表达,并具有良好的生物学活性;小鼠的免疫与攻毒试验结果表明,重组质粒pCAGGS-HA(ZJ245)首免后一周可检测到针对同源病毒ZJ245的低水平HI和VN抗体,加强免疫后抗体水平明显升高,HI抗体达到76.88、VN抗体达到152.5;同时针对异源病毒HLJ44也产生了较低水平的HI和VN抗体。106.0 EID50的同源病毒ZJ245攻毒时,与非免疫组小鼠相对比,重组质粒pCAGGS-HA(ZJ245)的免疫完全阻止了因病毒攻击而导致的小鼠体重下降以及肺脏和鼻甲内病毒的复制;106.0 EID50的异源病毒HLJ44攻毒时,相比非免疫组小鼠,质粒免疫组小鼠体重的下降比率、以及肺脏和鼻甲内的病毒滴度均显著降低(P<0.0001、P <0.001、P <0.05)。【结论】重组质粒pCAGGS-HA(ZJ245)能够有效表达病毒HA蛋白,免疫后可使小鼠获得完全抵抗ZJ245感染及部分抵御HLJ44感染的免疫保护力,表明重组质粒pCAGGS-HA(ZJ245)具有良好的免疫原性。
【Objective】This study aimed to construct the recombinant plasmid expressing HA gene of Eurasian avian-like H1 N1(EA H1 N1) swine influenza virus(SIV) and then evaluate its immunogenicity in mice. 【Method】HA gene of A/swine/Zhejiang/245/2013(H1 N1)(ZJ245) was amplified by RT-PCR, and inserted into an eukaryotic expression vector pCAGGS. The recombinant plasmid, designated as pCAGGS-HA(ZJ245), was transfected into 293 T cells, and the expressed HA protein was identified by indirect fluorescence assay(IFA) and Western blot. In order to evaluate the immunogenicity of the recombinant plasmid pCAGGSHA(ZJ245), sixteen six-week-old female BALB/c mice were immunized with 100 μg of the recombinant plasmid by intramuscular injection, and then were boosted once with a 3-week interval. Another group of sixteen mice received 100 μL of phosphate-buffered saline(PBS) were used as unvaccinated control. Serum samples were collected every week after prime and boost immunization in order to detect the hemagglutinin inhibition(HI) antibodies, virus neutralization(VN) antibodies, respectively. Two weeks after the boost immunization, pCAGGS-HA(ZJ245)-immunized and PBS-inoculated mice were intranasally challenged with 50 μL(106.0 EID50)of the homologous ZJ245 and heterologous A/swine/Heilongjiang/44/2009(H1 N1)(HLJ44), respectively. All mice per group were monitored daily for clinical signs of infection and body weight changes for two weeks. The mice that lost more than 25% of their initial body weight were euthanized on humane ground. On day 3 post-challenge three mice per group were euthanized and their organs including brain, nasal turbinate, lung, spleen and kidney were collected for virus titration in eggs. Immune efficacy of the recombinant plasmid p CAGGS-HA(ZJ245) was evaluated by body weight loss and virus replication titer in mice, respectively.【Result】The recombinant plasmid p CAGGS-HA(ZJ245) was constructed by inserting HA gene of ZJ245 virus and verified by restriction endonuclease analysis and plasmid sequencing. IFA and Western blot analysis confirmed that the HA protein could be correctly expressed by the recombinant plasmid pCAGGS-HA(ZJ245) and had a good biological activity in vitro. Immunization and challenge trial indicated that low levels of HI and VN antibody against the homologous ZJ245 virus were initially detected one week after the first immunization, and significantly increased after the second immunization, with the HI titer of 76.88 and the VN titer of152.5, respectively. Meanwhile, low levels of HI and VN antibodies against the heterologous HLJ44 virus were also detected.Compared with PBS-inoculated mice, the weight loss and viral replication rate of the pCAGGS-HA(ZJ245)-vaccinated mice when challenged with 106.0 EID50 of the homologous ZJ245 virus were completely inhibited. When challenged with 106.0 EID50 of the heterologous HLJ44 virus, the extent of weight loss and viral titer of the challenge virus detected in the p CAGGS-HA(ZJ245)-vaccinated mice were significantly lower than those in the PBS-inoculated mice(P<0.0001, P<0.001, P<0.05).【Conclusion】The recombinant plasmid p CAGGS-HA(ZJ245) could efficiently express HA protein, and provided complete protection for the immunized mice against the homologous ZJ245 virus infection and partial cross-protection against the heterologous HLJ44 virus infection, which indicated that the recombinant plasmid pCAGGS-HA(ZJ245) had good immunogenicity.
【Objective】This study aimed to construct the recombinant plasmid expressing HA gene of Eurasian avian-like H1 N1(EA H1 N1) swine influenza virus(SIV) and then evaluate its immunogenicity in mice. 【Method】HA gene of A/swine/Zhejiang/245/2013(H1 N1)(ZJ245) was amplified by RT-PCR, and inserted into an eukaryotic expression vector pCAGGS. The recombinant plasmid, designated as pCAGGS-HA(ZJ245), was transfected into 293 T cells, and the expressed HA protein was identified by indirect fluorescence assay(IFA) and Western blot. In order to evaluate the immunogenicity of the recombinant plasmid pCAGGSHA(ZJ245), sixteen six-week-old female BALB/c mice were immunized with 100 μg of the recombinant plasmid by intramuscular injection, and then were boosted once with a 3-week interval. Another group of sixteen mice received 100 μL of phosphate-buffered saline(PBS) were used as unvaccinated control. Serum samples were collected every week after prime and boost immunization in order to detect the hemagglutinin inhibition(HI) antibodies, virus neutralization(VN) antibodies, respectively. Two weeks after the boost immunization, pCAGGS-HA(ZJ245)-immunized and PBS-inoculated mice were intranasally challenged with 50 μL(106.0 EID50)of the homologous ZJ245 and heterologous A/swine/Heilongjiang/44/2009(H1 N1)(HLJ44), respectively. All mice per group were monitored daily for clinical signs of infection and body weight changes for two weeks. The mice that lost more than 25% of their initial body weight were euthanized on humane ground. On day 3 post-challenge three mice per group were euthanized and their organs including brain, nasal turbinate, lung, spleen and kidney were collected for virus titration in eggs. Immune efficacy of the recombinant plasmid p CAGGS-HA(ZJ245) was evaluated by body weight loss and virus replication titer in mice, respectively.【Result】The recombinant plasmid p CAGGS-HA(ZJ245) was constructed by inserting HA gene of ZJ245 virus and verified by restriction endonuclease analysis and plasmid sequencing. IFA and Western blot analysis confirmed that the HA protein could be correctly expressed by the recombinant plasmid pCAGGS-HA(ZJ245) and had a good biological activity in vitro. Immunization and challenge trial indicated that low levels of HI and VN antibody against the homologous ZJ245 virus were initially detected one week after the first immunization, and significantly increased after the second immunization, with the HI titer of 76.88 and the VN titer of152.5, respectively. Meanwhile, low levels of HI and VN antibodies against the heterologous HLJ44 virus were also detected.Compared with PBS-inoculated mice, the weight loss and viral replication rate of the pCAGGS-HA(ZJ245)-vaccinated mice when challenged with 106.0 EID50 of the homologous ZJ245 virus were completely inhibited. When challenged with 106.0 EID50 of the heterologous HLJ44 virus, the extent of weight loss and viral titer of the challenge virus detected in the p CAGGS-HA(ZJ245)-vaccinated mice were significantly lower than those in the PBS-inoculated mice(P<0.0001, P<0.001, P<0.05).【Conclusion】The recombinant plasmid p CAGGS-HA(ZJ245) could efficiently express HA protein, and provided complete protection for the immunized mice against the homologous ZJ245 virus infection and partial cross-protection against the heterologous HLJ44 virus infection, which indicated that the recombinant plasmid pCAGGS-HA(ZJ245) had good immunogenicity.
引文
[1]BROWN I.The epidemiology and evolution of influenza viruses in pigs.Veterinary Microbiology,2000,74(1/2):29-46.
[2]WEBBY R J,WEBSTER R G.Emergence of influenza A viruses.Philosophical Transactions of the Royal Society B Biological Sciences,2001,356(1416):1817-1828.doi:10.1098/rstb.2001.0997.
[3]BAUDON E,PEYRE M,PEIRIS M,COWLING B J.Epidemiological features of influenza circulation in swine populations:A systematic review and meta-analysis.PLoS ONE,2017,12(6):e179044.doi:10.1371/journal.pone.0179044.
[4]NEUMANN G,NODA T,KAWAOKA Y.Emergence and pandemic potential of swine-origin H1N1 influenza virus.Nature,2009,459(7249):931-939.doi:10.1038/nature08157.
[5]SCHULTZ U,FITCH W M,LUDWIG S,MANDLER J,SCHOLTISSEKC.Evolution of pig influenza viruses.Virology,1991,183(1):61-73.
[6]LIU J,BI Y,QIN K,FU G,YANG J,PENG J,MA G,LIU Q,PU J,TIAN F.Emergence of European avian influenza virus-like H1N1swine influenza A viruses in China.Journal of Clinical Microbiology,2009,47(8):2643-2646.doi:10.1128/JCM.00262-09.
[7]VINCENT A,AWADA L,BROWN I,CHEN H,CLAES F,DAUPHIN G,DONIS R,CULHANE M,HAMILTON K,LEWIS N,MUMFORD E,NGUYEN T,PARCHARIYANON S,PASICK J,PAVADE G,PEREDA A,PEIRIS M,SAITO T,SWENSON S,VANREETH K,WEBBY R,WONG F,CIACCI-ZANELLA J.Review of influenza A virus in swine worldwide:a call for increased surveillance and research.Zoonoses Public Health,2014,61(1):4-17.doi:10.1111/zph.12049.
[8]GUAN Y,SHORTRIDGE K F,KRAUSS S,LI P H,KAWAOKA Y,WEBSTER R G.Emergence of avian H1N1 influenza viruses in pigs in China.Journal of Virology,1996,70(11):8041-8046.
[9]YANG H,CHEN Y,QIAO C,HE X,ZHOU H,SUN Y,YIN H,MENG S,LIU L,ZHANG Q,KONG H,GU C,LI C,BU Z,KAWAOKA Y,CHEN H.Prevalence,genetics,and transmissibility in ferrets of Eurasian avian-like H1N1 swine influenza viruses.Proceedings of the National Academy of Sciences of the USA,2016,113(2):392-397.doi:10.1073/pnas.1522643113.
[10]CAMPITELLI L,DONATELLI I,FONI E,CASTRUCCI M R,FABIANI C,KAWAOKA Y,KRAUSS S,WEBSTER R G.Continued evolution of H1N1 and H3N2 influenza viruses in pigs in Italy.Virology,1997,232(2):310-318.doi:10.1006/viro.1997.8514.
[11]DE JONG J C,PACCAUD M F,DE RONDE-VERLOOP F M,HUFFELS N H,VERWEI C,WEIJERS T F,BANGMA P J,VANKREGTEN E,KERCKHAERT J A,WICKI F,WUNDERLI W.Isolation of swine-like influenza A(H1N1)viruses from man in Switzerland and the Netherlands.Annales de l'Institut Pasteur/Virologie,1988,139(4):429-437.
[12]WANG D Y,QI S X,LI X Y,GUO J F,TAN M J,HAN G Y,LIU YF,LAN Y,YANG L,HUANG W J,CHENG Y H,ZHAO X,BAI T,WANG Z,WEI H J,XIAO N,SHU Y L.Human infection with Eurasian avian-like influenza A(H1N1)virus,China.Emerging Infectious Diseases,2013,19(10):1709-1711.doi:10.3201/eid1910.130420.
[13]YANG H,QIAO C,TANG X,CHEN Y,XIN X,CHEN H.Human infection from avian-like influenza A(H1N1)viruses in pigs,China.Emerging Infectious Diseases,2012,18(7):1144-1146.doi:10.3201/eid1807.120009.
[14]SUI J,YANG D,QIAO C,XU H,XU B,WU Y,YANG H,CHEN Y,CHEN H.Protective efficacy of an inactivated Eurasian avian-like H1N1 swine influenza vaccine against homologous H1N1 and heterologous H1N1 and H1N2 viruses in mice.Vaccine,2016,34(33):3757-3763.doi:10.1016/j.vaccine.2016.06.009.
[15]RUAN B,WEN F,GONG X,LIU X M,WANG Q,YU L X,WANGS Y,ZHANG P,YANG H M,SHAN T L,ZHENG H,ZHOU Y J,TONG W,GAO F,TONG G Z,YU H.Protective efficacy of a high-growth reassortant H1N1 influenza virus vaccine against the European avian-like H1N1 swine influenza virus in mice and pigs.Veterinary Microbiology,2018,222:75-84.doi:10.1016/j.vetmic.2018.07.003.
[16]LIU L,LU J,ZHOU J,LI Z,ZHANG H,WANG D,SHU Y.Construction and comparison of different source neuraminidase candidate vaccine strains for human infection with Eurasian avian-like influenza H1N1 virus.Microbes and Infection,2017,19(12):635-640.doi:10.1016/j.micinf.2017.08.004.
[17]WU Y,YANG D,XU B,LIANG W,SUI J,CHEN Y,YANG H,CHEN H,WEI P,QIAO C.Immune efficacy of an adenoviral vector-based swine influenza vaccine against antigenically distinct H1N1 strains in mice.Antiviral Research,2017,147:29-36.doi:10.1016/j.antiviral.2017.09.009.
[18]SHVARTSMAN D E,KOTLER M,TALL R D,ROTH M G,HENISY I.Differently anchored influenza hemagglutinin mutants display distinct interaction dynamics with mutual rafts.Journal of Cell Biology,2003,163(4):879-888.doi:10.1083/jcb.200308142.
[19]TREGONING J S,KINNEAR E.Using plasmids as DNA vaccines for infectious diseases.Microbiology Spectrum,2014,2(6):1-16.doi:10.1128/microbiolspec.PLAS-0028-2014.
[20]徐汇洋,许榜丰,陈艳,隋金钰,杨焕良,尹航,杨大为,乔传玲,陈化兰.一株H1N1猪流感病毒的进化分析与分子特征[J].中国农业科学,2015(15):3071-3078.doi:10.3864/j.issn.0578-1752.2015.15.018.XU H Y,XU B F,CHEN Y,SUI J Y,YANG H L,YIN H,YANG DW,QIAO C L,CHEN H L.Phylogenetic analysis and molecular characteristics of an H1N1subtype swine influenza virus.Scientia Agricultura Sinica,2015(15):3071-3078.(in Chinese).doi:10.3864/j.issn.0578-1752.2015.15.018.
[21]CHEN Y,ZHANG J,QIAO C,YANG H,ZHANG Y,XIN X,CHENH.Co-circulation of pandemic 2009 H1N1,classical swine H1N1 and avian-like swine H1N1 influenza viruses in pigs in China.Infection,Genetics and Evolution,2013,13:331-338.doi:10.1016/j.meegid.2012.09.021.
[22]PENSAERT M,OTTIS K,VANDEPUTTE J,KAPLAN M M,BACHMANN P A.Evidence for the natural transmission of influenza A virus from wild ducts to swine and its potential importance for man.Bulletin of the World Health Organ,1981,59(1):75-78.
[23]DUCATEZ M F,HAUSE B,STIGGER-ROSSER E,DARNELL D,CORZO C,JULEEN K,SIMONSON R,BROCKWELL-STAATS C,RUBRUM A,WANG D,WEBB A,CRUMPTON J C,LOWE J,GRAMER M,WEBBY R J.Multiple reassortment between pandemic(H1N1)2009 and endemic influenza viruses in pigs,United States.Emerging Infectious Diseases,2011,17(9):1624-1629.doi:10.3201/eid1709.110338.
[24]QIAO C,LIU L,YANG H,CHEN Y,XU H,CHEN H.Novel triple reassortant H1N2 influenza viruses bearing six internal genes of the pandemic 2009/H1N1 influenza virus were detected in pigs in China.Journal of Clinical Virology,2014,61(4):529-534.doi:10.1016/j.jcv.2014.10.014.
[25]WATSON S J,LANGAT P,REID S M,LAM TT,COTTEN M,KELLY M,VAN REETH K,QIU Y,SIMON G,BONIN E,FONI E,CHIAPPONI C,LARSEN L,HJULSAGER C,MARKOWSKA-DANIEL I,URBANIAK K,DüRRWALD R,SCHLEGEL M,HUOVILAINEN A,DAVIDSON I,DáNá,LOEFFEN W,EDWARDS S,BUBLOT M,VILA T,MALDONADO J,VALLS L,ESNIP3 CONSORTIUM,BROWN IH,PYBUS OG,KELLAM P.Molecular epidemiology and evolution of influenza viruses circulating within European swine between 2009 and 2013.Journal of Virology,2015,89(19):9920-9931.doi:10.1128/JVI.00840-15.
[26]RAJ?O D S,CHEN H,PEREZ D R,SANDBULTE M R,GAUGERP C,LOVING C L,SHANKS G D,VINCENT A.Vaccine-associated enhanced respiratory disease is influenced by Haemagglutinin and Neuraminidase in whole inactivated influenza virus vaccines.Journal of General Virology,2016,97(7):1489-1499.doi:10.1099/jgv.0.000468.
[27]PILLET S,KOBASA D,MEUNIER I,GRAY M,LADDY D,WEINER D B,VON MESSLING V,KOBINGER G P.Cellular immune response in the presence of protective antibody levels correlates with protection against 1918 influenza in ferrets.Vaccine,2011,29(39):6793-6801.doi:10.1016/j.vaccine.2010.12.059.
[28]JIANG Y,YU K,ZHANG H,ZHANG P,LI C,TIAN G,LI Y,WANG X,GE J,BU Z,CHEN H.Enhanced protective efficacy of H5subtype avian influenza DNA vaccine with codon optimized HA gene in a pCAGGS plasmid vector.Antiviral Research,2007,75(3):234-241.doi:10.1016/j.antiviral.2007.03.009.
[29]PING X,HU W,XIONG R,ZHANG X,TENG Z,DING M,LI L,CHANG C,XU K.Generation of a broadly reactive influenza H1antigen using a consensus HA sequence.Vaccine,2018,36(32 Pt B):4837-4845.doi:10.1016/j.vaccine.2018.06.048.
[30]MURPHY B R,CLEMENTS M L.The systemic and mucosal immune response of humans to influenza A virus.Current Topics in Microbiology and Immunology,1989,146:107-116.
[31]DE VLEESCHAUWER A R,VAN POUCKE S G,KARASIN A I,OLSEN CW,VAN REETH K.Cross-protection between antigenically distinct H1N1 swine influenza viruses from Europe and North America.Influenza Other Respiratory Viruses,2011,5(2):115-122.doi:10.1111/j.1750-2659.2010.00164.x.
[32]BRAUCHER D R,HENNINGSON J N,LOVING C L,VINCENT AL,KIM E,STEITZ J,GAMBOTTO A A,KEHRLI ME J R.Intranasal vaccination with replication-defective adenovirus type 5encoding influenza virus hemagglutinin elicits protective immunity to homologous challenge and partial protection to heterologous challenge in pigs.Clinical and Vaccine Immunology,2012,19(11):1722-1729.doi:10.1128/CVI.00315-12.
[2]WEBBY R J,WEBSTER R G.Emergence of influenza A viruses.Philosophical Transactions of the Royal Society B Biological Sciences,2001,356(1416):1817-1828.doi:10.1098/rstb.2001.0997.
[3]BAUDON E,PEYRE M,PEIRIS M,COWLING B J.Epidemiological features of influenza circulation in swine populations:A systematic review and meta-analysis.PLoS ONE,2017,12(6):e179044.doi:10.1371/journal.pone.0179044.
[4]NEUMANN G,NODA T,KAWAOKA Y.Emergence and pandemic potential of swine-origin H1N1 influenza virus.Nature,2009,459(7249):931-939.doi:10.1038/nature08157.
[5]SCHULTZ U,FITCH W M,LUDWIG S,MANDLER J,SCHOLTISSEKC.Evolution of pig influenza viruses.Virology,1991,183(1):61-73.
[6]LIU J,BI Y,QIN K,FU G,YANG J,PENG J,MA G,LIU Q,PU J,TIAN F.Emergence of European avian influenza virus-like H1N1swine influenza A viruses in China.Journal of Clinical Microbiology,2009,47(8):2643-2646.doi:10.1128/JCM.00262-09.
[7]VINCENT A,AWADA L,BROWN I,CHEN H,CLAES F,DAUPHIN G,DONIS R,CULHANE M,HAMILTON K,LEWIS N,MUMFORD E,NGUYEN T,PARCHARIYANON S,PASICK J,PAVADE G,PEREDA A,PEIRIS M,SAITO T,SWENSON S,VANREETH K,WEBBY R,WONG F,CIACCI-ZANELLA J.Review of influenza A virus in swine worldwide:a call for increased surveillance and research.Zoonoses Public Health,2014,61(1):4-17.doi:10.1111/zph.12049.
[8]GUAN Y,SHORTRIDGE K F,KRAUSS S,LI P H,KAWAOKA Y,WEBSTER R G.Emergence of avian H1N1 influenza viruses in pigs in China.Journal of Virology,1996,70(11):8041-8046.
[9]YANG H,CHEN Y,QIAO C,HE X,ZHOU H,SUN Y,YIN H,MENG S,LIU L,ZHANG Q,KONG H,GU C,LI C,BU Z,KAWAOKA Y,CHEN H.Prevalence,genetics,and transmissibility in ferrets of Eurasian avian-like H1N1 swine influenza viruses.Proceedings of the National Academy of Sciences of the USA,2016,113(2):392-397.doi:10.1073/pnas.1522643113.
[10]CAMPITELLI L,DONATELLI I,FONI E,CASTRUCCI M R,FABIANI C,KAWAOKA Y,KRAUSS S,WEBSTER R G.Continued evolution of H1N1 and H3N2 influenza viruses in pigs in Italy.Virology,1997,232(2):310-318.doi:10.1006/viro.1997.8514.
[11]DE JONG J C,PACCAUD M F,DE RONDE-VERLOOP F M,HUFFELS N H,VERWEI C,WEIJERS T F,BANGMA P J,VANKREGTEN E,KERCKHAERT J A,WICKI F,WUNDERLI W.Isolation of swine-like influenza A(H1N1)viruses from man in Switzerland and the Netherlands.Annales de l'Institut Pasteur/Virologie,1988,139(4):429-437.
[12]WANG D Y,QI S X,LI X Y,GUO J F,TAN M J,HAN G Y,LIU YF,LAN Y,YANG L,HUANG W J,CHENG Y H,ZHAO X,BAI T,WANG Z,WEI H J,XIAO N,SHU Y L.Human infection with Eurasian avian-like influenza A(H1N1)virus,China.Emerging Infectious Diseases,2013,19(10):1709-1711.doi:10.3201/eid1910.130420.
[13]YANG H,QIAO C,TANG X,CHEN Y,XIN X,CHEN H.Human infection from avian-like influenza A(H1N1)viruses in pigs,China.Emerging Infectious Diseases,2012,18(7):1144-1146.doi:10.3201/eid1807.120009.
[14]SUI J,YANG D,QIAO C,XU H,XU B,WU Y,YANG H,CHEN Y,CHEN H.Protective efficacy of an inactivated Eurasian avian-like H1N1 swine influenza vaccine against homologous H1N1 and heterologous H1N1 and H1N2 viruses in mice.Vaccine,2016,34(33):3757-3763.doi:10.1016/j.vaccine.2016.06.009.
[15]RUAN B,WEN F,GONG X,LIU X M,WANG Q,YU L X,WANGS Y,ZHANG P,YANG H M,SHAN T L,ZHENG H,ZHOU Y J,TONG W,GAO F,TONG G Z,YU H.Protective efficacy of a high-growth reassortant H1N1 influenza virus vaccine against the European avian-like H1N1 swine influenza virus in mice and pigs.Veterinary Microbiology,2018,222:75-84.doi:10.1016/j.vetmic.2018.07.003.
[16]LIU L,LU J,ZHOU J,LI Z,ZHANG H,WANG D,SHU Y.Construction and comparison of different source neuraminidase candidate vaccine strains for human infection with Eurasian avian-like influenza H1N1 virus.Microbes and Infection,2017,19(12):635-640.doi:10.1016/j.micinf.2017.08.004.
[17]WU Y,YANG D,XU B,LIANG W,SUI J,CHEN Y,YANG H,CHEN H,WEI P,QIAO C.Immune efficacy of an adenoviral vector-based swine influenza vaccine against antigenically distinct H1N1 strains in mice.Antiviral Research,2017,147:29-36.doi:10.1016/j.antiviral.2017.09.009.
[18]SHVARTSMAN D E,KOTLER M,TALL R D,ROTH M G,HENISY I.Differently anchored influenza hemagglutinin mutants display distinct interaction dynamics with mutual rafts.Journal of Cell Biology,2003,163(4):879-888.doi:10.1083/jcb.200308142.
[19]TREGONING J S,KINNEAR E.Using plasmids as DNA vaccines for infectious diseases.Microbiology Spectrum,2014,2(6):1-16.doi:10.1128/microbiolspec.PLAS-0028-2014.
[20]徐汇洋,许榜丰,陈艳,隋金钰,杨焕良,尹航,杨大为,乔传玲,陈化兰.一株H1N1猪流感病毒的进化分析与分子特征[J].中国农业科学,2015(15):3071-3078.doi:10.3864/j.issn.0578-1752.2015.15.018.XU H Y,XU B F,CHEN Y,SUI J Y,YANG H L,YIN H,YANG DW,QIAO C L,CHEN H L.Phylogenetic analysis and molecular characteristics of an H1N1subtype swine influenza virus.Scientia Agricultura Sinica,2015(15):3071-3078.(in Chinese).doi:10.3864/j.issn.0578-1752.2015.15.018.
[21]CHEN Y,ZHANG J,QIAO C,YANG H,ZHANG Y,XIN X,CHENH.Co-circulation of pandemic 2009 H1N1,classical swine H1N1 and avian-like swine H1N1 influenza viruses in pigs in China.Infection,Genetics and Evolution,2013,13:331-338.doi:10.1016/j.meegid.2012.09.021.
[22]PENSAERT M,OTTIS K,VANDEPUTTE J,KAPLAN M M,BACHMANN P A.Evidence for the natural transmission of influenza A virus from wild ducts to swine and its potential importance for man.Bulletin of the World Health Organ,1981,59(1):75-78.
[23]DUCATEZ M F,HAUSE B,STIGGER-ROSSER E,DARNELL D,CORZO C,JULEEN K,SIMONSON R,BROCKWELL-STAATS C,RUBRUM A,WANG D,WEBB A,CRUMPTON J C,LOWE J,GRAMER M,WEBBY R J.Multiple reassortment between pandemic(H1N1)2009 and endemic influenza viruses in pigs,United States.Emerging Infectious Diseases,2011,17(9):1624-1629.doi:10.3201/eid1709.110338.
[24]QIAO C,LIU L,YANG H,CHEN Y,XU H,CHEN H.Novel triple reassortant H1N2 influenza viruses bearing six internal genes of the pandemic 2009/H1N1 influenza virus were detected in pigs in China.Journal of Clinical Virology,2014,61(4):529-534.doi:10.1016/j.jcv.2014.10.014.
[25]WATSON S J,LANGAT P,REID S M,LAM TT,COTTEN M,KELLY M,VAN REETH K,QIU Y,SIMON G,BONIN E,FONI E,CHIAPPONI C,LARSEN L,HJULSAGER C,MARKOWSKA-DANIEL I,URBANIAK K,DüRRWALD R,SCHLEGEL M,HUOVILAINEN A,DAVIDSON I,DáNá,LOEFFEN W,EDWARDS S,BUBLOT M,VILA T,MALDONADO J,VALLS L,ESNIP3 CONSORTIUM,BROWN IH,PYBUS OG,KELLAM P.Molecular epidemiology and evolution of influenza viruses circulating within European swine between 2009 and 2013.Journal of Virology,2015,89(19):9920-9931.doi:10.1128/JVI.00840-15.
[26]RAJ?O D S,CHEN H,PEREZ D R,SANDBULTE M R,GAUGERP C,LOVING C L,SHANKS G D,VINCENT A.Vaccine-associated enhanced respiratory disease is influenced by Haemagglutinin and Neuraminidase in whole inactivated influenza virus vaccines.Journal of General Virology,2016,97(7):1489-1499.doi:10.1099/jgv.0.000468.
[27]PILLET S,KOBASA D,MEUNIER I,GRAY M,LADDY D,WEINER D B,VON MESSLING V,KOBINGER G P.Cellular immune response in the presence of protective antibody levels correlates with protection against 1918 influenza in ferrets.Vaccine,2011,29(39):6793-6801.doi:10.1016/j.vaccine.2010.12.059.
[28]JIANG Y,YU K,ZHANG H,ZHANG P,LI C,TIAN G,LI Y,WANG X,GE J,BU Z,CHEN H.Enhanced protective efficacy of H5subtype avian influenza DNA vaccine with codon optimized HA gene in a pCAGGS plasmid vector.Antiviral Research,2007,75(3):234-241.doi:10.1016/j.antiviral.2007.03.009.
[29]PING X,HU W,XIONG R,ZHANG X,TENG Z,DING M,LI L,CHANG C,XU K.Generation of a broadly reactive influenza H1antigen using a consensus HA sequence.Vaccine,2018,36(32 Pt B):4837-4845.doi:10.1016/j.vaccine.2018.06.048.
[30]MURPHY B R,CLEMENTS M L.The systemic and mucosal immune response of humans to influenza A virus.Current Topics in Microbiology and Immunology,1989,146:107-116.
[31]DE VLEESCHAUWER A R,VAN POUCKE S G,KARASIN A I,OLSEN CW,VAN REETH K.Cross-protection between antigenically distinct H1N1 swine influenza viruses from Europe and North America.Influenza Other Respiratory Viruses,2011,5(2):115-122.doi:10.1111/j.1750-2659.2010.00164.x.
[32]BRAUCHER D R,HENNINGSON J N,LOVING C L,VINCENT AL,KIM E,STEITZ J,GAMBOTTO A A,KEHRLI ME J R.Intranasal vaccination with replication-defective adenovirus type 5encoding influenza virus hemagglutinin elicits protective immunity to homologous challenge and partial protection to heterologous challenge in pigs.Clinical and Vaccine Immunology,2012,19(11):1722-1729.doi:10.1128/CVI.00315-12.
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