口蹄疫病毒感染中乳鼠epsilon干扰素mRNA转录水平的实时荧光定量PCR检测
|
摘要
利用3~4日龄的乳鼠作为研究对象,在口蹄疫病毒(FMDV)MYA/98株攻毒后第0、5、10小时及濒临死亡期(第15小时)收集其肠道组织并提取RNA,用实时荧光定量PCR检测乳鼠IFN-ε表达量在感染前后及不同时间段的变化。结果显示,IFN-ε表达量在乳鼠感染FMDV前后以及不同时间段并没有发生明显的变化。本研究结果为进一步了解Ⅰ型干扰素的不同亚型及其在抗口蹄疫病毒感染中的作用提供了实验数据。
The suckling mice aging from 3—4 days-old were employed and the intestinal tissue was collected at 0 h,5 h,10 h,and 15 h after the mice were challenged with foot-and-mouth disease virus(FMDV)MYA/98 strain.Real-time quantitative PCR was utilized to detect the IFN-ε mRNA transcription levels of suckling mouse before and after challenge.The results showed that IFN-ε expression would not alter between uninfected and acutely infected intestinal tissue or between the difference time-points postinfection.These results provided some experimental data for further understanding the subtypes of typeⅠinterferon and their role in anti-infection of FMDV infection.
The suckling mice aging from 3—4 days-old were employed and the intestinal tissue was collected at 0 h,5 h,10 h,and 15 h after the mice were challenged with foot-and-mouth disease virus(FMDV)MYA/98 strain.Real-time quantitative PCR was utilized to detect the IFN-ε mRNA transcription levels of suckling mouse before and after challenge.The results showed that IFN-ε expression would not alter between uninfected and acutely infected intestinal tissue or between the difference time-points postinfection.These results provided some experimental data for further understanding the subtypes of typeⅠinterferon and their role in anti-infection of FMDV infection.
引文
[1]PESTKA S,KRAUSE C D,WALTER M R.Interferons,interferon-like cytokines,and their receptors[J].Immunological Reviews,2004,202:8-32.
[2]FUNG K Y,MANGAN N E,CUMMING H,et al.Interferon-εprotects the female reproductive tract from viral and bacterial infection[J].Science,2013,339(6123):1088-1092.
[3]GARCIA-MINAMBRES A,EID S G,MANGAN N E,et al.Interferon epsilon promotes HIV restriction at multiple steps of viral replication[J].Immunology and Cell Biology,2017,95(5):478-483.
[4]ABDULHAQQ S A,ZORRILLA C,KANG G,et al.HIV-1-negative female sex workers sustain high cervical IFN epsilon,low immune activation,and low expression of HIV-1-required host genes[J].Mucosal Immunology,2016,9(4):1027-1038.
[5]RODRIGUEZ PULIDO M,SAIZ M.Molecular mechanisms of foot-and-mouth disease virus targeting the host antiviral response[J].Frontiers in Cellular and Infection Microbiology,2017,7:252.
[6]USHARANI J,PARK S Y,CHO E J,et al.Antiviral activity of ovine interferon tau 4 against foot-and-mouth disease virus[J].Antiviral Research,2017,143:134-141.
[7]HABIELA M,SEAGO J,PEREZ-MARTIN E,et al.Laboratory animal models to study foot-and-mouth disease:a review with emphasis on natural and vaccineinduced immunity[J].Journal of General Virology,2014,95(11):2329-2345.
[8]RUBIO N,PALOMO M,ALCAMI A.Interferon-α/βgenes are up-regulated in murine brain astrocytes after infection with Theiler’s murine encephalomyelitis virus[J].Journal of Interferon&Cytokine Research,2010,30(4):253-262.
[9]POTT J,MAHLAKOIV T,MORDSTEIN M,et al.IFN-γdetermines the intestinal epithelial antiviral host defense[J].Proceedings of the National Academic Sciences of the United States of America,2011,108(19):7944-7949.
[10]HARDY M P,OWCZAREK C M,JERMIIN L S,et al.Characterization of the typeⅠinterferon locus and identification of novel genes[J].Genomics,2004,84:331-345.
[11]SANG Y,ROWLAND R R R,HESSE R A,et al.Differential expression and activity of the porcine typeⅠinterferon family[J].Physiological Genomics,2010,42(2):248-258.
[12]GUO Y,GAO M,BAO J,et al.Molecular cloning and characterization of a novel bovine IFN-epsilon[J].Gene,2015,558:25-30.
[13]YANG L,XU L,LI Y,et al.Molecular and functional characterization of canine interferon-epsilon[J].Journal of Interferon&Cytokine Research,2013,33(12):760-768.
[14]DEMERS A,KANG G,MA F,et al.The mucosal expression pattern of interferon-εin rhesus macaques[J].Journal of Leukocyte Biology,2014,96(6):1101-1107.
[15]CHOO S W,RAYKO M,TAN T K,et al.Pangolin genomes and the evolution of mammalian scales and immunity[J].Genome Research,2016,26(10):1312-1322.
[16]ROBERTSEN B.The role of typeⅠinterferons in innate and adaptive immunity against viruses in Atlantic salmon[J].Developmental&Comparative Immunology,2017,pii:S0145-305X(17)30076-9.
[17]BOASSO A,ROYLE C M,DOUMAZOS S,et al.Overactivation of plasmacytoid dendritic cells inhibits antiviral T-cell responses:a model for HIV immunopathogenesis[J].Blood,2011,118:5152-5162.
[18]BOSINGER S E,LI Q,GORDON S N,et al.Global genomic analysis reveals rapid control of a robust innate response in SIV-infected sooty mangabeys[J].The Journal of Clinical Investigation,2009,119(12):3556-3572.
[19]DIAZ-SAN SEGUNDO F,WEISS M,PEREZ-MARTIN E,et al.Antiviral activity of bovine typeⅢinterferon against foot-and-mouth disease virus[J].Virology,2011,413(2):283-292.
[20]KIM S M,KIM S K,PARK J H,et al.A recombinant adenovirus bicistronically expressing porcine interferon-αand interferon-γenhances antiviral effects against foot-and-mouth disease virus[J].Antiviral Research,2014,104(1):52-58.
[21]MORAES M P,DE LOS SANTOS T,KOSTER M,et al.Enhanced antiviral activity against foot-and-mouth disease virus by a combination of typeⅠandⅡporcine interferons[J].Journal of Virology,2007,81(13):7124-7135.
[22]ROODGAR M,ROSS C T,TARARA R,et al.Gene expression and TB pathogenesis in rhesus macaques:TR4,CD40,CD40L,FAS(CD95),and TNF are host genetic markers in peripheral blood mononuclear cells that are associated with severity of TB lesions[J].Infection&Genetic Evolution,2015,36:396-409.
[23]HERMANT P,FRANCIUS C,CLOTMAN F,et al.IFN-εis constitutively expressed by cells of the reproductive tract and is inefficiently secreted by fibroblasts and cell lines[J].PLo S One,2013,8(8):e71320.
[2]FUNG K Y,MANGAN N E,CUMMING H,et al.Interferon-εprotects the female reproductive tract from viral and bacterial infection[J].Science,2013,339(6123):1088-1092.
[3]GARCIA-MINAMBRES A,EID S G,MANGAN N E,et al.Interferon epsilon promotes HIV restriction at multiple steps of viral replication[J].Immunology and Cell Biology,2017,95(5):478-483.
[4]ABDULHAQQ S A,ZORRILLA C,KANG G,et al.HIV-1-negative female sex workers sustain high cervical IFN epsilon,low immune activation,and low expression of HIV-1-required host genes[J].Mucosal Immunology,2016,9(4):1027-1038.
[5]RODRIGUEZ PULIDO M,SAIZ M.Molecular mechanisms of foot-and-mouth disease virus targeting the host antiviral response[J].Frontiers in Cellular and Infection Microbiology,2017,7:252.
[6]USHARANI J,PARK S Y,CHO E J,et al.Antiviral activity of ovine interferon tau 4 against foot-and-mouth disease virus[J].Antiviral Research,2017,143:134-141.
[7]HABIELA M,SEAGO J,PEREZ-MARTIN E,et al.Laboratory animal models to study foot-and-mouth disease:a review with emphasis on natural and vaccineinduced immunity[J].Journal of General Virology,2014,95(11):2329-2345.
[8]RUBIO N,PALOMO M,ALCAMI A.Interferon-α/βgenes are up-regulated in murine brain astrocytes after infection with Theiler’s murine encephalomyelitis virus[J].Journal of Interferon&Cytokine Research,2010,30(4):253-262.
[9]POTT J,MAHLAKOIV T,MORDSTEIN M,et al.IFN-γdetermines the intestinal epithelial antiviral host defense[J].Proceedings of the National Academic Sciences of the United States of America,2011,108(19):7944-7949.
[10]HARDY M P,OWCZAREK C M,JERMIIN L S,et al.Characterization of the typeⅠinterferon locus and identification of novel genes[J].Genomics,2004,84:331-345.
[11]SANG Y,ROWLAND R R R,HESSE R A,et al.Differential expression and activity of the porcine typeⅠinterferon family[J].Physiological Genomics,2010,42(2):248-258.
[12]GUO Y,GAO M,BAO J,et al.Molecular cloning and characterization of a novel bovine IFN-epsilon[J].Gene,2015,558:25-30.
[13]YANG L,XU L,LI Y,et al.Molecular and functional characterization of canine interferon-epsilon[J].Journal of Interferon&Cytokine Research,2013,33(12):760-768.
[14]DEMERS A,KANG G,MA F,et al.The mucosal expression pattern of interferon-εin rhesus macaques[J].Journal of Leukocyte Biology,2014,96(6):1101-1107.
[15]CHOO S W,RAYKO M,TAN T K,et al.Pangolin genomes and the evolution of mammalian scales and immunity[J].Genome Research,2016,26(10):1312-1322.
[16]ROBERTSEN B.The role of typeⅠinterferons in innate and adaptive immunity against viruses in Atlantic salmon[J].Developmental&Comparative Immunology,2017,pii:S0145-305X(17)30076-9.
[17]BOASSO A,ROYLE C M,DOUMAZOS S,et al.Overactivation of plasmacytoid dendritic cells inhibits antiviral T-cell responses:a model for HIV immunopathogenesis[J].Blood,2011,118:5152-5162.
[18]BOSINGER S E,LI Q,GORDON S N,et al.Global genomic analysis reveals rapid control of a robust innate response in SIV-infected sooty mangabeys[J].The Journal of Clinical Investigation,2009,119(12):3556-3572.
[19]DIAZ-SAN SEGUNDO F,WEISS M,PEREZ-MARTIN E,et al.Antiviral activity of bovine typeⅢinterferon against foot-and-mouth disease virus[J].Virology,2011,413(2):283-292.
[20]KIM S M,KIM S K,PARK J H,et al.A recombinant adenovirus bicistronically expressing porcine interferon-αand interferon-γenhances antiviral effects against foot-and-mouth disease virus[J].Antiviral Research,2014,104(1):52-58.
[21]MORAES M P,DE LOS SANTOS T,KOSTER M,et al.Enhanced antiviral activity against foot-and-mouth disease virus by a combination of typeⅠandⅡporcine interferons[J].Journal of Virology,2007,81(13):7124-7135.
[22]ROODGAR M,ROSS C T,TARARA R,et al.Gene expression and TB pathogenesis in rhesus macaques:TR4,CD40,CD40L,FAS(CD95),and TNF are host genetic markers in peripheral blood mononuclear cells that are associated with severity of TB lesions[J].Infection&Genetic Evolution,2015,36:396-409.
[23]HERMANT P,FRANCIUS C,CLOTMAN F,et al.IFN-εis constitutively expressed by cells of the reproductive tract and is inefficiently secreted by fibroblasts and cell lines[J].PLo S One,2013,8(8):e71320.