Poly(I:C)对PRRSV诱导的抗病毒免疫反应的影响  

Effects of Poly(I:C)on antiviral immune response induced by porcine reproductive and respiratory syndrome virus

作　　者：王冬梅[1] 崔春晓[1] 冯亚琼[1] 张莹莹[1] 郭嘉[1] 杜东颖 夏平安[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《中国兽医学报》2016年第2期211-215,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31172346)

摘　　要：为了研究聚肌苷胞对猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus，PRRSV）诱导的抗病毒免疫反应的影响，本研究采用200个TCID50的PRRSV感染猪肺巨噬细胞（porcine alveolarmacro—phage，PAM），吸附1h后用100μg／L的聚肌苷胞（poly（I：C））处理细胞，分别培养12、24、36、48h后收集细胞及上清，同时设PRRSV感染对照组、poly（I：C）刺激对照组和健康细胞对照组，用实验室已经建立的Real—timeqPCR方法对poly（I：C）刺激PRRSV感染组和PRRSV感染对照组中PRRSV复制水平及各组PAM细胞中IFN-α、TNF-αmRNA转录水平变化进行定量分析。结果显示，PRRSV在感染PAM细胞后12～24h期间可促进PAM细胞IFN—αmRNA的转录，36～48h期间抑制PAM细胞IFN—αmRNA的转录；TNF—αmRNA转录水平在12、48h后略微升高，在24、36h后均略微下降。Poly（I：C）处理PAM细胞后IFN—α、TNF-αmRNA转录水平均上调。Poly（I：C）＋PRRSV组IFN—αmRNA的转录水平与PRRSV组相比在12～36h均显著升高，在48h后则显著下调。Poly（I：C）＋PRRSV组TNF—αmRNA转录水平与PRRSV组相比，12～36h后均升高，在12h升高较显著，48h后则显著下调。结果表明，PRRSV感染PAM细胞经Poly（I：C）刺激后IFN—α、TNF-αmRNA转录水平在12～36h后显著升高，从而抑制了PRRSV在PAM细胞内的复制。To study the effects of poly（I：C） on antiviral immune response induced by porcine reproductive and respiratory syndrome virus （PRRSV）, porcine alveolar macrophage （PAM） was prepared and cultured with PRRSV solution 200 TCID50. After inoculated 1 h,cells were treated with 100μg/L poly（I：C）. Then cells and supernatants were collected after treated 12,24,36 and 48 h, respectively. The 1 virus,IFN-α and TNF-α mRNA in the PAM cells were quantitated by real-time qPCR including poly（I：C） stimulation of PRRSV infected PAM cells group,PRRSV infection control group,poly（I.C）stimulated cells control group and the cell control group. The results showed that mRNA level of IFN-α in PAW was improved at 12 and 24 h after PRRSV infection,and mR- NA level of IFN-α was inhibited at 36 and 48 h;TNF-α mRNA level was increased slightly at 12 and 48 h post-infection, and was decreased slightly at 24 and 36 h. mRNA levels of IFN-α and TNF-α of PAM cells treated with poly（I：C） were both improved at 12 and 48 h;IFN-a and TNF-α mRNA levels of poly（I.C） stimulation of PRRSV infected PAM cells were both up-regulated at 12 and 36 h compared with PRRSV infection control group. So after stimulating with poly （I：C） ,the mRNA levels of IFN-α and TNF-α mRNA in the PAM cells infected by RRSV were im proved（P〈0.05） ,which inhibited the replication of PRRSV in PAM cell.

关 键 词：猪繁殖与呼吸综合征病毒 猪肺泡巨噬细胞 IFN-Α 肿瘤坏死因子 Poly(I:C) 

分 类 号：S852.65[农业科学—基础兽医学]