SHP1在PRRSV诱导的抗病毒免疫反应中的作用
发布时间：2018-11-29 13:17
【摘要】：为了探究SHP1在猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)诱导的抗病毒免疫中的作用,本研究采用200个TCID50的PRRSV感染猪肺巨噬细胞(porcine alveolar macrophage,PAM),分别培养12,24,36,48h后收集细胞及上清,分别设正常细胞对照组,聚肌胞(polyinosinic-polycytidylic acid,polyI:C)刺激对照组,脂多糖(lipopolysaccharide,LPS)刺激对照组。采用ELISA方法检测PRRSV感染的PAM培养上清中SHP1的蛋白表达情况。用实验室已经建立的实时荧光定量PCR方法对PRRSV感染组和各个对照组PAM细胞中IRF3、IRF7、TRAF6、NF-κB、SHP1和IFN-β的mRNA转录水平变化进行定量分析。结果显示:激活的SHP1对PRRSV感染后PAM中IRF7、NF-kB和IFN-β的转录有促进作用,对IRF3和TRAF6的转录有抑制作用。LPS及Poly(I:C)刺激PRRSV感染的PAM细胞均促进了IRF3、IRF7、TRAF6、NF-κB、SHP1和IFN-β的转录。结果表明,激活的SHP1会通过TLR介导的信号通路或未知通路来调节PRRSV诱导的抗病毒免疫反应水平。LPS和Poly(I:C)刺激PRRSV感染的PAM细胞能够增强抗病毒细胞因子的转录水平。
[Abstract]:To investigate the role of SHP1 in porcine reproductive and respiratory syndrome virus (porcine reproductive and respiratory syndrome virus,PRRSV) -induced antiviral immunity, 200 TCID50 PRRSV were used to infect porcine pulmonary macrophages (porcine alveolar macrophage,PAM). The cells and supernatants were collected after 48 hours of culture, and were divided into normal cell control group, polymyocyte-stimulated control group (polyinosinic-polycytidylic acid,polyI:C) and lipopolysaccharide (lipopolysaccharide,LPS) stimulated control group. ELISA method was used to detect the expression of SHP1 protein in the supernatant of PAM infected with PRRSV. The changes of mRNA transcription levels of IRF3,IRF7,TRAF6,NF- 魏 B, SHP1 and IFN- 尾 in PAM cells of PRRSV infected group and control group were quantitatively analyzed by real-time fluorescence quantitative PCR assay. The results showed that activated SHP1 promoted the transcription of IRF7,NF-kB and IFN- 尾 in PAM after PRRSV infection, and inhibited the transcription of IRF3 and TRAF6. LPS and Poly (I: C) stimulated PAM cells infected with PRRSV to promote IRF3,IRF7,TRAF6,. Transcription of NF- 魏 B, SHP1 and IFN- 尾. The results showed that activated SHP1 regulated the level of antiviral immune response induced by PRRSV through TLR mediated signaling pathway or unknown pathway. LPS and Poly (I / C) stimulated PRRSV infected PAM cells to enhance the transcription level of antiviral cytokines.
【作者单位】： 河南农业大学牧医工程学院;
【基金】：国家自然科学基金资助项目(31172346)
【分类号】：S852.65
本文编号：2365096
[Abstract]:To investigate the role of SHP1 in porcine reproductive and respiratory syndrome virus (porcine reproductive and respiratory syndrome virus,PRRSV) -induced antiviral immunity, 200 TCID50 PRRSV were used to infect porcine pulmonary macrophages (porcine alveolar macrophage,PAM). The cells and supernatants were collected after 48 hours of culture, and were divided into normal cell control group, polymyocyte-stimulated control group (polyinosinic-polycytidylic acid,polyI:C) and lipopolysaccharide (lipopolysaccharide,LPS) stimulated control group. ELISA method was used to detect the expression of SHP1 protein in the supernatant of PAM infected with PRRSV. The changes of mRNA transcription levels of IRF3,IRF7,TRAF6,NF- 魏 B, SHP1 and IFN- 尾 in PAM cells of PRRSV infected group and control group were quantitatively analyzed by real-time fluorescence quantitative PCR assay. The results showed that activated SHP1 promoted the transcription of IRF7,NF-kB and IFN- 尾 in PAM after PRRSV infection, and inhibited the transcription of IRF3 and TRAF6. LPS and Poly (I: C) stimulated PAM cells infected with PRRSV to promote IRF3,IRF7,TRAF6,. Transcription of NF- 魏 B, SHP1 and IFN- 尾. The results showed that activated SHP1 regulated the level of antiviral immune response induced by PRRSV through TLR mediated signaling pathway or unknown pathway. LPS and Poly (I / C) stimulated PRRSV infected PAM cells to enhance the transcription level of antiviral cytokines.
【作者单位】： 河南农业大学牧医工程学院;
【基金】：国家自然科学基金资助项目(31172346)
【分类号】：S852.65
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