两株生物学特性不同的新城疫病毒La Sota毒株诱导DF-1细胞天然免疫反应的比较

The different host innate immune responses of the DF-1 cells induced by two different La Sota strains of Newcastle disease virus

为探讨两株不同生物学特性的新城疫病毒La Sota毒株(La Sota参考毒株AV1615;La Sota突变株P2M1)引起的宿主天然免疫反应的差异,将两个毒株分别感染鸡胚成纤维细胞(DF-1),Real-time PCR及Western blot检测TLR3、TLR7、MDA5等受体的mRNA及蛋白表达;ELISA及免疫荧光检测细胞核转录因子IRF3的含量及其磷酸化水平;蛋白芯片检测40种细胞因子的分泌水平,并用ELISA验证部分与炎症、抗病毒等相关的细胞因子在细胞上清中的含量差异。结果显示,La Sota参考毒株AV1615感染细胞后,诱导产生的TLR3、TLR7、MDA5的mRNA及蛋白含量显著高于La Sota突变株P2M1(P<0.05)。La Sota感染细胞后12 h,IRF3及其磷酸化水平、细胞上清中大部分细胞因子(31/40)的表达高于P2M1感染组。结果说明,两株La Sota毒株在诱导DF-1细胞的天然免疫反应方面存在差异,这可能是La Sota P2M1在鸡胚中更容易增殖的原因。

In order to compare the different host innate immune responses induced by two strains of Newcastle disease viruses（ La Sota reference strain AV1615 and mutation strain P2M1） in chicken embryo fibroblasts cell line（ DF-1）,the cells were inoculated with the two strains of La Sota,respectively,and the pattern recognition receptor（ TLR3,TLR7 and MDA5）,transcription factor（ IRF3） and cytokines were detected. The results showed that the mRNA and protein expression levels of TLR3,TLR7 and MDA5 induced by La Sota AV1615 were significantly higher than those induced by La Sota P2M1（ P〈 0. 05）. The expression levels of IRF3,the phosphorylated IRF3 and most of the cytokines（ 31/40） in the supernatant of DF-1 cells in La Sota AV1615 group were also higher than those in La Sota P2M1 challenging group at 12 h after infection. These results indicated that different host innate immune responses were induced by these two viruses,which may explain why La Sota P2M1 had higher titer in chick embryo.