PRRSV感染对猪肺泡巨噬细胞IFITM3基因转录的影响  

EFFECT OF PRRSV INFECTION ON IFITM3 TRANSCRIPTION IN PORCINE ALVEOLAR MACROPHAGES

作　　者：李玉明[1] 武专昌[1] 刘珂[1] 马改妮[1] 魏建超[1] 邵东华[1] 李蓓蓓[1] 邱亚峰[1] 石元元[1] 马志永[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2018年第1期7-12,共6页Chinese Journal of Animal Infectious Diseases

基　　金：973项目计划课题(2014CB542702);国家自然科学基金(31502046)

摘　　要：干扰素诱导跨膜蛋白3(interferon inducible transmembrane proteins 3,IFITM3)作为抗病毒蛋白可以抑制多种病毒的感染,本研究针对猪IFITM3基因设计特异性引物,构建重组质粒并作为阳性标准品,建立了检测IFITM3基因的SYBR Green Ⅰ荧光定量RT-PCR方法。结果显示,该方法最低检出下限为10~2 copies/μL,且线性关系好(R^2≥0.997),敏感性高,特异性强,重复性好,批内、批间变异系数均小于3%。将猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)感染猪肺泡巨噬细胞(porcine alveolar macrophages,PAM)细胞,在不同感染时间对细胞中IFITM3 mRNA进行检测。结果显示,PRRSV感染早期IFITM3转录水平保持不变,在感染后转录水平逐渐升高。IFITM3 mRNA在PRRSV感染PAM过程中转录变化,可能是PRRSV宿主免疫逃逸的机制之一。As an antivirus protein, interferon inducible transmembrane protein 3 （IFITM3） inhibits a broad spectrum of virus infections. In this study, the speciflc primers were designed based on the genes of IFITM3 and GAPDH to detect IFITM3 expression in PRRSV-infected porcine alveolar macrophages （PAMs）. The recombinant plasmids containing the target genes were constructed as a standard control and real time SYBR Green I RT-PCR method was developed for detection of IFITM3 expression. The results showed that the detection limit of this method was 102 copies/μL with good linear relationship, specificity, sensitivity and repeatability. Moreover, the variation coefflcient of the method was less than 3%. We used this method to detect the IFITM3 mRNA levels in PRRSV-infected PAMs. The results revealed that the IFITM3 mRNA levels showed no signiflcant difference at the early stage of infection but increased significantly at the late stage of infection in PRRSV-infected PAMs as compared with mock-infected PAMs, demonstrating that the changes of IFITM3 mRNA levels in PRRSV-infected PAMs might be related to PRRSV infection as well as immune evasion.

关 键 词：猪干扰素诱导跨膜蛋白3 猪繁殖与呼吸综合征病毒 SYBR Green Ⅰ荧光定量PCR 猪肺泡巨噬细胞 

分 类 号：S852.44[农业科学—基础兽医学]