TLR2介导鸡毒支原体脂质相关膜蛋白诱导DF-1细胞表达IL-1β的研究

发布时间：2018-04-03 15:30

本文选题：鸡毒支原体　切入点：脂质相关膜蛋白(LAMPs)　出处：《中国预防兽医学报》2017年02期

【摘要】：Toll样受体2(TLR2)在鸡毒支原体(MG)诱导天然免疫反应机制中的作用尚未明确。为鉴定TLR2在MG脂质相关膜蛋白(LAMPs)诱导细胞表达IL-1β中的作用,本研究通过RT-PCR和ELISA方法检测LAMPs刺激后DF-1细胞中IL-1β的m RNA和蛋白水平,结果显示LAMPs可以诱导DF-1细胞分泌IL-1β,并确定最佳刺激时间为6 h,最佳刺激剂量为1μg/m L。此外,通过构建TLR2重组表达质粒p CMV-HA-TLR2,并将该质粒转染DF-1细胞进行过表达及通过抗体封闭TLR2的两种处理方式,处理后采用LAMPs刺激细胞。经检测显示:TLR2过表达处理组的IL-1βm RNA和蛋白水平显著升高,而抗体处理组的IL-1βm RNA和蛋白水平下调。结果表明,LAMPs能够与DF-1细胞膜上的TLR2受体结合,有效的诱导DF-1细胞表达并分泌IL-1β,而且IL-1β的释放可能与天然免疫系统的经典通路即NF-κB信号通路调控有关。本研究为阐述MG的致病机制提供相关实验依据。
[Abstract]:The role of Toll-like receptor 2-TLR2 in the mechanism of innate immune response induced by Mycoplasma gallus is unclear.In order to investigate the role of TLR2 in the expression of IL-1 尾 in MG lipide-associated membrane protein (lamps), the m RNA and protein levels of IL-1 尾 in DF-1 cells stimulated by LAMPs were detected by RT-PCR and ELISA.The results showed that LAMPs could induce DF-1 cells to secrete IL-1 尾, and the optimal stimulation time was 6 h and the optimal stimulation dose was 1 渭 g / mL.In addition, the recombinant TLR2 expression plasmid pCMV-HA-TLR2 was constructed and transfected into DF-1 cells for overexpression and blocking of TLR2 by antibody. After treatment, the cells were stimulated by LAMPs.The results showed that the IL-1 尾 m RNA and protein levels were significantly increased in the overexpression treatment group, while the IL-1 尾 m RNA and protein levels were down-regulated in the antibody treatment group.The results showed that lamps could bind to TLR2 receptor on DF-1 cell membrane and induce DF-1 cells to express and secrete IL-1 尾 effectively. Moreover, the release of IL-1 尾 may be related to the regulation of NF- 魏 B signaling pathway, which is the classical pathway of innate immune system.This study provides experimental basis for elucidating the pathogenesis of MG.
【作者单位】：东北农业大学动物医学学院;湖北工业大学生物工程与食品学院;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物支原体病团队;东北农业大学资源与环境学院;吉林农业大学动物科学技术学院;
【基金】：防治畜禽支原体病中药制剂的研究及应用黑龙江省高校科技成果产业化前期研发培育项目(1254CGZH16) 自噬在TLR2介导鸡支原体炎性损伤中的作用机制研究兽医生物技术国家重点实验室开放课题基金(SKLVBF201409)
【分类号】：S852.62

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