H9N2亚型禽流感病毒对鸡的免疫抑制机制
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摘要
通过评价H9N2亚型禽流感病毒(AIVs)感染对于鸡新城疫和传染性支气管炎活疫苗免疫效力的影响,研究了H9N2 AIVs的免疫抑制机理。结果显示,H9N2 AIVs感染可降低鸡对鸡新城疫和传染性支气管炎活疫苗的抗体应答,但与对照组相比差异并不显著;而流式分析结果显示,与对照组相比H9N2 AIVs感染可显著降低鸡外周血中CD4+T淋巴细胞和CD8+T淋巴细胞的比率(P<0.05);抗原特异性的T淋巴细胞增殖试验结果显示,与对照组相比H9N2 AIVs感染鸡的外周血T淋巴细胞增殖活性显著下降(P<0.05);另外,H9N2 AIV感染还导致鸡血清中细胞因子IFN-γ和IL-4应答水平的下降,其中IFN-γ的应答水平与对照组相比差异显著(P<0.05)。说明H9N2 AIVs感染可抑制鸡的免疫应答,其中对细胞免疫应答的抑制更为明显。
The influence of H9 N2 subtype avian influenza viruses(H9 N2 AIVs) infection on the immune efficacy of newcastle disease virus(NDV) and infectious bronchitis virus(IBV) vaccine was assessed to determine the immune suppressive mechanism for H9 N2 virus. The results showed that the infection of H9 N2 virus reduced the antibody titers to the live vaccine of NDV and IBV,but this difference was not significant compared with the control group. However,the flow cytometric analysis of peripheral blood lymphocyte subpopulations showed that H9 N2 virus infection resulted in a significantly decrease in CD4 + T lymphocyte and CD8+ T lymphocyte proportion,and the antigen specific proliferation ability of peripheral blood T lymphocyte in H9 N2 virus infection group was also obviously suppressed compared with that of the control group. Moreover,the level of cytokines IFN-γ and IL-4 in serum of H9 N2 virus infection chickens was also lower than that of control group,and the difference of IFN-γ was significant. The results suggested that H9 N2 AIVs infection could suppress the immune response to vaccine in chickens,and the cellular immune response was more significantly suppressed.
The influence of H9 N2 subtype avian influenza viruses(H9 N2 AIVs) infection on the immune efficacy of newcastle disease virus(NDV) and infectious bronchitis virus(IBV) vaccine was assessed to determine the immune suppressive mechanism for H9 N2 virus. The results showed that the infection of H9 N2 virus reduced the antibody titers to the live vaccine of NDV and IBV,but this difference was not significant compared with the control group. However,the flow cytometric analysis of peripheral blood lymphocyte subpopulations showed that H9 N2 virus infection resulted in a significantly decrease in CD4 + T lymphocyte and CD8+ T lymphocyte proportion,and the antigen specific proliferation ability of peripheral blood T lymphocyte in H9 N2 virus infection group was also obviously suppressed compared with that of the control group. Moreover,the level of cytokines IFN-γ and IL-4 in serum of H9 N2 virus infection chickens was also lower than that of control group,and the difference of IFN-γ was significant. The results suggested that H9 N2 AIVs infection could suppress the immune response to vaccine in chickens,and the cellular immune response was more significantly suppressed.
引文
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[24]KASH J C,BASLER C F,GARCIA-SASTRE A,et al.Global host immune response:pathogenesis and transcriptional profiling of type A influenza viruses expressing the hemagglutinin and neuraminidase genes from the 1918 pandemic virus[J].J Virol,2004,78:9499-9511.
[25]WATANABE T,KISO M,FUKUYAMA S,et al.Characterization of H7N9 influenza A viruses isolated from humans[J].Nature,2013,501:551-555.
[26]SMED-SORENSEN A,CHALOUNI C,CHATTERJEE B,et al.Influenza A virus infection of human primary dendritic cells impairs their ability to cross-present antigen to CD8 T cells[J].PLo S Pathog,2012,8:e1002572.
[27]KODIHALLI S,SIVANANDAN V,NAGARAJA K V,et al.Effect of avian influenza virus infection on the phagocytic function of systemic phagocytes and pulmonary macrophages of turkeys[J].Avian Dis,1994,38:93-102.
[28]谷长勤,罗玲,胡薛英,等.H9N2亚型禽流感病毒诱导蛋鸡免疫器官细胞凋亡的动态变化[J].中国兽医学报,2008,28(1):12-14.
[29]袁建琴,高斌战,梁新华.H9亚型禽流感病毒对鸡免疫器官影响的研究试验[J].兽药市场指南,2008(12):39-10.
[2]陈伯伦,张泽纪,陈伟斌.禽流感研究:I鸡A型流感病毒分离与血清学初步鉴定[J].中国兽医杂志,1994,22(10):3-5.
[3]LIU J K,OKAZAKI H,OZAKI Y,et al.H9N2 influenza viruses prevalent in poultry in China are phylogenetically distinct from A/quail/Hong Kong/G1/97 presumed to be the donor of the internal protein genes of the H5N1 Hong Kong/97 virus[J].Avian Pathol,2003,32:551-560.
[4]LIU H X,LIU J,CHENG D,et al.Phylogenetic analysis of the hemagglutinin genes of twenty-six avian influenza viruses of subtype H9N2 isolated from chickens in China during 1996-2001[J].Avian Dis,2003,47:116-127.
[5]LI C K,YU G,TIAN D,et al.Evolution of H9N2 influenza viruses from domestic poultry in Mainland China[J].Virology,2005,340:70-83.
[6]ZHIRNOV O P,IKIZLER M R,WRIGHT P F.Cleavage of influenza a virus hemagglutinin in human respiratory epithelium is cell associated and sensitive to exogenous antiproteases[J].J Virol,2002,76:8682-8689.
[7]朱明霞,牛玉娟,马海营,等.规模化肉鸡场常见呼吸道病原体感染状况调查[J].山东农业大学学报(自然科学版),2015,46(6):861-864.
[8]KAREEM E,HASSAN,AHMED A,et al.Experimental co-infection of infectious bronchitis and low pathogenic avian influenza H9N2 viruses in commercial broiler chickens[J].Res Vet Sci,2017,115:356-362.
[9]毕英佐.发鸡呼吸道堵塞的疾病探究[J].北方牧业,2016(14):23.
[10]NILI H,ASASI K.Natural cases and an experimental study of H9N2 avian influenza in commercial broiler chickens of Iran[J].Avian Pathol,2002,31:247-252.
[11]仇保丰,刘武杰,胡顺林,等.混合感染的多种亚型禽流感病毒的纯化与鉴定[J].微生物学报,2010,50(1):107-112.
[12]ROUSSAN D A,HADDAD R,KHAWALDEH G.Molecular survey of avian respiratory pathogens in commercial broiler chicken flocks with respiratory diseases in Jordan[J].Poult Sci,2008,87:444-448.
[13]LEI T,YING L,JIN F,et al.Prediction and identification of novel IBV S1 protein derived CTL epitopes in chicken[J].Vaccine,2016,34:380-386.
[14]MAREK W,ESAM T A,JANA S,et al.Major histocompatibility complex(MHC)class i and MHC class ii proteins:conformational plasticity in antigen presentation[J].Front Microbiol,2017,8:292.
[15]PAUL W E.Interleukin-4:a prototypic immunoregulatory lymphokine[J].Blood,1991,77:1859-1870.
[16]MEEK B,SPEIJER D,DE JONG P T,et al.The ocular humoral immuneresponse in health and disease[J].Prog Retin Eye Res,2003,22(3):391-415.
[17]ARAI K I,LEE F,MIYAJIMA A,et al.Cytokines:coordinators of immune and inflammatory responses[J].Annu Rev Biochem,1990,59:783-836.
[18]LOWENTHAL J W,YORK J J,O NEIL J J,et al.In vivo effects of chicken IFN-during infection with Eimeria[J].J Interferon Cytokine Res,1997,17(9):551-558.
[19]WEINING K C,SCHULTZ U,MUNSTER U,et al.Biological properties of recombinant chicken interferon gamma[J].Eur J Immunol,1996,26:2440-2447.
[20]SONG K D,LILLEHOJ H S,CHOI K D,et al.Expression and functional characterization of recombinant chicken interferon gamma[J].Vet Immunol Immunopathol,1997,58:321-333.
[21]DE JONG M D,SIMMONS C P,THANH T T,et al.Fatal outcome of human influenza A(H5N1)is associated with high viral load and hypercytokinemia[J].Nat Med,2006,12:1203-1207.
[22]CHEUNG C Y,POON L L,LAU A S,et al.Induction of proinflammatory cytokines in human macrophages by influenza A(H5N1)viruses:a mechanism for the unusual severity of human disease[J].Lancet,2002,360:1831-1837.
[23]TAUBENBERGER J K,REID A H,KRAFFT A E,et al.Initial genetic characterization of the 1918‘Spanish’influenza virus[J].Science,1997,275:1793-1796.
[24]KASH J C,BASLER C F,GARCIA-SASTRE A,et al.Global host immune response:pathogenesis and transcriptional profiling of type A influenza viruses expressing the hemagglutinin and neuraminidase genes from the 1918 pandemic virus[J].J Virol,2004,78:9499-9511.
[25]WATANABE T,KISO M,FUKUYAMA S,et al.Characterization of H7N9 influenza A viruses isolated from humans[J].Nature,2013,501:551-555.
[26]SMED-SORENSEN A,CHALOUNI C,CHATTERJEE B,et al.Influenza A virus infection of human primary dendritic cells impairs their ability to cross-present antigen to CD8 T cells[J].PLo S Pathog,2012,8:e1002572.
[27]KODIHALLI S,SIVANANDAN V,NAGARAJA K V,et al.Effect of avian influenza virus infection on the phagocytic function of systemic phagocytes and pulmonary macrophages of turkeys[J].Avian Dis,1994,38:93-102.
[28]谷长勤,罗玲,胡薛英,等.H9N2亚型禽流感病毒诱导蛋鸡免疫器官细胞凋亡的动态变化[J].中国兽医学报,2008,28(1):12-14.
[29]袁建琴,高斌战,梁新华.H9亚型禽流感病毒对鸡免疫器官影响的研究试验[J].兽药市场指南,2008(12):39-10.