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为探究猪圆环病毒2型(PCV2)感染对TLR3信号通路及炎性细胞因子的影响,本研究将30只6周龄ICR雌鼠随机分成两组(感染组和对照组),腹腔接种PCV2b/YJ株,在接种后第7 d、14 d、21 d、28 d和35 d采集小鼠外周血及脾组织。以临床观察、脾组织PCV2基因拷贝、小鼠血清抗体效价、IL-10和TNF-α的mRNA水平等指标评估感染模型;荧光定量PCR检测小鼠脾组织TLR3、TRIF、IFN-β、Mx1的mRNA水平,western blot检测TLR3蛋白表达水平。结果显示:PCV2感染期间感染组小鼠无可见的临床症状,第7 d至35 d感染组小鼠脾组织PCV2基因拷贝数呈逐渐降低趋势,PCV2的IPMA血清抗体在感染后第7 d转阳、35 d达到最高,IL-10 mRNA在感染后第14 d起升高、维持高水平;TNF-αmRNA在感染后第21 d和28 d升高。在感染后第7 d和14 d感染组小鼠TLR3、TRIF mRNA和TLR3蛋白表达水平显著升高;IFN-βmRNA在感染后第7 d具有显著升高过程;Mx1 mRNA在感染后第14 d时有上调过程。本实验在建立PCV2b/YJ亚临床感染ICR小鼠模型的基础上,体内实验表明PCV2感染小鼠激活了TLR3信号通路。本研究为利用敲除小鼠模型阐明TLR3参与PCV2感染引发的免疫应答机制奠定了基础。
In order to investigate the effect of porcine circovirus type 2 (PCV2) infection on TLR3 signaling pathway and inflammatory cytokines, 30 female 6-week-old ICR mice were randomly divided into two groups (infected group and control group), intraperitoneally inoculated with PCV2b / YJ strain. The peripheral blood and spleen tissues of mice were collected on the 7th, 14th, 21st, 28th and 35th day after inoculation. The model of infection was evaluated by clinical observation, PCV2 gene copy of spleen, serum antibody titers of mice, mRNA level of IL-10 and TNF-α, TLR3, TRIF, IFN-β, Mx1 The mRNA level of TLR3 was detected by western blot. The results showed that there was no visible clinical symptoms in infected mice during PCV2 infection, and the copy number of PCV2 gene in spleen tissue of mice infected with PCV2 gradually decreased from the 7th day to the 35th day after infection. IPMA serum antibody of PCV2 turned positive on the 7th day , Reached the highest level on the 35th day. The IL-10 mRNA level increased from the 14th day after infection to a high level. The TNF-αmRNA increased on the 21st and 28th day after infection. The expression of TLR3, TRIF mRNA and TLR3 protein were significantly increased in infected mice on the 7th day and the 14th day after infection. The IFN-βmRNA increased significantly on the 7th day after infection. On the 14th day after infection, Upward process. In this study, PCV2b / YJ subclinical infection in ICR mouse model was established based on in vivo experiments show that PCV2 infected mice activate the TLR3 signaling pathway. This study laid the foundation for the use of a knockout mouse model to elucidate the mechanism by which TLR3 is involved in the immune response triggered by PCV2 infection.