H5N1禽流感病毒感染雏鸭免疫器官TGF-β1 mRNA转录及病理损伤的变化
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摘要
为了探索鹅源H5N1禽流感病毒(AIV)感染对雏鸭胸腺、脾脏、法氏囊的病理损伤及对TGF-β1mRNA转录的影响,将120只21日龄雏鸭随机分为AIV感染组(I)和对照组(C),应用RT-PCR和免疫组织化学技术结合病理学检测方法,对上述免疫器官的细胞凋亡数量、病理形态变化以及TGF-β1 mRNA转录进行了系统研究。结果显示,雏鸭感染AIV后第3～5天,免疫器官的病理损伤最为明显,脾脏、胸腺和法氏囊细胞凋亡数量分别在病毒感染后第1～5天、第3～5天显著(P<0.05)或极显著(P<0.01)高于对照雏鸭。TGF-β1 mRNA转录均在病毒感染后第1～5天较对照雏鸭明显(P<0.05)升高。上述研究结果表明,鹅源H5N1亚型AIV感染雏鸭后的初期,不但免疫器官第的淋巴细胞凋亡明显,而且TGF-β1 mRNA转录也明显升高;鹅源H5N1亚型AIV既是引起感染雏鸭免疫功能下降、呈现免疫抑制的主要因素,也是导致病毒感染雏鸭死亡的主要原因之一。
order to explore the pathological damage of thymus,spleen,bursal and the transcriptional changes of TGF-β1 mRNA in ducklings infected with goose-derived H5 N1 avian influenza virus,and to explain its mechanism,120 ducklings aged 21 days were randomly divided into AIV infection group(I)and control group(C).The number of apoptosis, the above immune organs were detected by the RT-PCR method,immunohistochemistry technique and pathological examination.In result,the pathological damages in immune organs of ducklings infected with AIV were the most obvious on day 3 to 5 after the infection.The number of apoptosis in spleens,thymus and bursa of ducklings infected with AIV were significantly higher than that in the control group on day 1 to 5 and on day 3 to 5 after infection differently.And the expression of TGF-β1 mRNA in the immune organs was significantly higher than that in the control group on day 1 to 5 after the infection.In conclusion,at the initial stage after the infection,the number of lymphocytes apoptosis were obvious in immune organs of ducklings infected with AIV,but also the transcriptions of TGF-β1 mRNA was significantly increased.The changes were as well as the main factor of decrease immune function of infected ducklings,which were also one of the main causes of death of infected ducklings.
order to explore the pathological damage of thymus,spleen,bursal and the transcriptional changes of TGF-β1 mRNA in ducklings infected with goose-derived H5 N1 avian influenza virus,and to explain its mechanism,120 ducklings aged 21 days were randomly divided into AIV infection group(I)and control group(C).The number of apoptosis, the above immune organs were detected by the RT-PCR method,immunohistochemistry technique and pathological examination.In result,the pathological damages in immune organs of ducklings infected with AIV were the most obvious on day 3 to 5 after the infection.The number of apoptosis in spleens,thymus and bursa of ducklings infected with AIV were significantly higher than that in the control group on day 1 to 5 and on day 3 to 5 after infection differently.And the expression of TGF-β1 mRNA in the immune organs was significantly higher than that in the control group on day 1 to 5 after the infection.In conclusion,at the initial stage after the infection,the number of lymphocytes apoptosis were obvious in immune organs of ducklings infected with AIV,but also the transcriptions of TGF-β1 mRNA was significantly increased.The changes were as well as the main factor of decrease immune function of infected ducklings,which were also one of the main causes of death of infected ducklings.
引文
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[21] XU S H,NGUYEN T T,QUAN S L,et al.Transforming growth Factor-β1-induced apoptosis in podocytes via extracellular-signal-regulated kinase-mammalian target of rapamycin complex 1-NADPH oxidase 4 axis[J].Journal of Biological Chemistry,2015,290:30830-30842
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[23]于学武,吕晓萍,刘超男,等.雏鸭感染鹅源H5N1亚型禽流感病毒后免疫器官T细胞免疫功能及IL-2 mRNA转录变化[J].畜牧兽医学报,2014,45(9):1497-1504.YU Xue-wu,LU Xiao-ping,LIU Chao-nan,et al.Changes of T lymphocytes proliferation function,quantity of TANAE+cells and transcription of IL-2 mRNA in immune organs of dicks infected with H5N1 subtype avian influenza virus isolated from geese[J].Acta Veterinaria et Zootechnica Sinica,2014,45(9):1497-1504.(in Chinese)
[24] HIROSHI T,TAKUYA N,MASAFUMI T.Functional heterogeneity of human effector CD8+T cells[J].Blood,2011,1182(10):343251.
[25]刘晓霞,李凤鸣,陈育民,等.转化生长因子β1对人外周血T淋巴细胞体外增殖及活化的影响[J].实用医学杂志,2009,25(20):3373-3375.LIU Xiao-xia,LI Feng-ming,CHEN Yu-min,et al.Effects of transforming growth factorβ1 on proliferation and activation of T lymphocytes in peripheral blood in vitro[J].The Journal of Practical Medicine,2009,25(20):3373-3375.
[26] AFSANA I,MOHAMMED E C,YUKA K,et al.Sustained anti-inflammatory effects of TGF-β1 on microglia/macrophages[J].Biochimica et Biophysica Acta(BBA)-Molecular Basis of Disease,2018,1864:721-734.
[27] SUN C,FU B Q,GAO Y F,et al.TGF-β1 down-regulation of NKG2D/DAP10 and 2B4/SAP expression on human NK cells contributes to HBV persistence[J].Plos Pathogens,2012,1371(10):1002594.
[2] URSZULA J,ANNA S,AGNIESZKA S,et al.A biosensor based on electroactive dipyrromethene-Cu(II)layer deposited onto gold electrodes for the detection of antibodies against avian influenza virus type H5N1 in hen sera[J].Analytical and Bioanalytical Chemistry,2015,407:7807-7814.
[3]蔡仁龙,林少娟,曾文娟.高致病性禽流感的防治[J].福建畜牧兽医,2013,35(2):56-57.CAO Ren-long,LIN Shao-juan,ZENG Wen-juan.Prevention and control of highly pathogenic avian influenza[J].Fujian Journal of Animal Husbandry and Veterinary Medicine,2013,35(2):56-57.
[4]丁晴微,于杨,崔鹏飞,等.湖南湖北两省H6亚型禽流感病毒表面基因的序列分析[J].基因组学与应用生物学,2011,30(3):282-287.DING Qing-wei,YU Yang,CUI Peng-fei,et al.Surface genes analysis of H6 subtype avian influenza virus isolated from Hunan and Hubei provinces[J].Genomics and Applied Biology,2011,30(3):282-287.(in Chinese)
[5] LI C J,BU Z G,CHEN H L.Avian in uenza vaccines against H5N1‘bird flu’[J].Trends in Biotechnology,2014,32(3):147-156.
[6]王伯沄,李玉松,黄高昇,等.病理学技术[M].北京:人民卫生出版社,2000.WANG Bo-yun,LI Yu-song,HUANG Gao-sheng,et al.Pathologic Technique[M].Beijing:People's Medical Publishing House,2000.(in Chinese)
[7] EDWARD W A B,SUSAN J M,CLIVE S.Role of apoptosis and cytokines in influenza virus morbidity[J].FEMS Microbiology Reviews,2005,29:837-850.
[8]王军政,王开,张宁宁,等.鸭源H5N5和H5N1亚型禽流感病毒BALB/c鼠感染试验[J].中国兽医学报,2015,35(12):1948-1953.WANG Jun-zheng,WANG Kai,ZHANG Ning-ning,et al.Infection experiment of BALB/c mice with duck-originated H5N5 and H5N1 subtype avian influenza virus[J].Chinese Journal of Veterinary Science,2015,35(12):1948-1953.(in Chinese)
[9] ZHANG C F,YANG Y T,ZHOU X W,et al.Highly pathogenic avian influenza A virus H5N1 NS1 protein induces caspase-dependent apoptosis in human alveolar basal epithelial cells[J].Virology Journal,2010,7(51):1743-422X.
[10]张卓.H5N1亚型禽流感病毒感染雏鸡的病理学变化及其发生机制[D].哈尔滨:东北农业大学,2007.ZHANG Zhuo.Pathobiology and pathogenesis of chickens infected with HSN1 avian influenza virus[D].Harbin:Northeast Agricultural University,2007.(in Chinese)
[11] WU H B,PENG X R,XU L H,et al.Characterization of a novel highly pathogenic H5N2 avian influenza virus isolated from a duck in eastern China[J].Archives of Virology,2014,159:3377-3383.
[12]刘超男,刘明,张云,等.H5N1亚型禽流感病毒的鸭致病性研究[J].中国农业科学,2006,39(2):412-417.LIU Chao-nan,LIU Ming,ZHANG Yun,et al.Pathogenicity of H5N1 influenza virus for ducks[J].Scientia Agricultura Sinica,2006,39(2):412-417.(in Chinese)
[13]许静,宋家升,苏胜杰,等.H5N1亚型禽流感病毒对鸭的致病性研究[J].中国预防兽医学报,2010,32(4):301-303.XU Jing,SONG Jia-sheng,SU Sheng-jie,et al.Virulence and replication of H5N1 avian influenza viruses in domestic ducks[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(4):301-303.(in Chinese)
[14] CUI Z,HU J,HE L,et al.Differential immune response of mallard duck peripheral blood mononuclear cells to two highly pathogenic avian influenza H5N1 viruses with distinct pathogenicity in mallard ducks[J].Archives of Virology,2014,159(2):339-343.
[15] WEI L M,JIAO P R,SONG Y F,et al.Host immune responses of ducks infected with H5N1 highly pathogenic avian influenza viruses of different pathogenicities[J].Veterinary Microbiology,2013,166(3-4):386-393.
[16] CARAN C,JAMIE W,ERICA S,et al.Differences in pathogenicity, response to vaccination, and innate immune responses in different types of ducks infected with a virulent H5N1 highly pathogenic avian influenza virus from vietnam[J].American Association of Avian Pathologists,2012,56(3):479-487.
[17] PASCALE M,CLAIRE D,AURELIE O,et al.Differential cellular gene expression in duck trachea infected with a highly or low pathogenic H5N1 avian influenza virus[J].Virology Journal,2013,10:279.
[18] SIMON B,ADAM J.K,JOHN B,et al.H5N1 infection causes rapid mortality and high cytokine lev els in chickens compared to ducks[J].Virus Research,2014,185:23-31.
[19] PEI X D,ZHAI Y F,ZHANG H H.Influenza virus H1N1induced apoptosis of mouse astrocytes and the effect on protein expression[J].Asian Pacific Journal of Tropical Medicine,2014,7(7):572-575.
[20]张瑞莉,刘明,张卓,等.鹅源H5N1亚型禽流感病毒感染雏鸡免疫器官细胞凋亡的研究[J].中国预防兽医学报,2011,33(3):177-180.ZHANG Rui-li,LIU Ming,ZHANG Zhuo,et al.Apoptosis in the immune organs of chickens infected with H5N1 avian influenza virus derived from goose[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(3):177-180.(in Chinese)
[21] XU S H,NGUYEN T T,QUAN S L,et al.Transforming growth Factor-β1-induced apoptosis in podocytes via extracellular-signal-regulated kinase-mammalian target of rapamycin complex 1-NADPH oxidase 4 axis[J].Journal of Biological Chemistry,2015,290:30830-30842
[22] WANG L X,WEN W H,YUAN J L,et al.Immunotherapy for human renal cell carcinoma by adoptive transfer of autologous transforming growth factorβ–insensitive CD8+T Cells[J].Clinical Cancer Research,2009,10:1158/1078-0432.
[23]于学武,吕晓萍,刘超男,等.雏鸭感染鹅源H5N1亚型禽流感病毒后免疫器官T细胞免疫功能及IL-2 mRNA转录变化[J].畜牧兽医学报,2014,45(9):1497-1504.YU Xue-wu,LU Xiao-ping,LIU Chao-nan,et al.Changes of T lymphocytes proliferation function,quantity of TANAE+cells and transcription of IL-2 mRNA in immune organs of dicks infected with H5N1 subtype avian influenza virus isolated from geese[J].Acta Veterinaria et Zootechnica Sinica,2014,45(9):1497-1504.(in Chinese)
[24] HIROSHI T,TAKUYA N,MASAFUMI T.Functional heterogeneity of human effector CD8+T cells[J].Blood,2011,1182(10):343251.
[25]刘晓霞,李凤鸣,陈育民,等.转化生长因子β1对人外周血T淋巴细胞体外增殖及活化的影响[J].实用医学杂志,2009,25(20):3373-3375.LIU Xiao-xia,LI Feng-ming,CHEN Yu-min,et al.Effects of transforming growth factorβ1 on proliferation and activation of T lymphocytes in peripheral blood in vitro[J].The Journal of Practical Medicine,2009,25(20):3373-3375.
[26] AFSANA I,MOHAMMED E C,YUKA K,et al.Sustained anti-inflammatory effects of TGF-β1 on microglia/macrophages[J].Biochimica et Biophysica Acta(BBA)-Molecular Basis of Disease,2018,1864:721-734.
[27] SUN C,FU B Q,GAO Y F,et al.TGF-β1 down-regulation of NKG2D/DAP10 and 2B4/SAP expression on human NK cells contributes to HBV persistence[J].Plos Pathogens,2012,1371(10):1002594.