STAT3介导猪扁桃体上皮细胞感染红斑丹毒丝菌的作用机制研究

Tonsillar mucosal epithelium was the first barrier preventing intrusions of E. rhusiopathiae. However, whether the route of mucosal infection involves epithelial cells was related to bacterial virulence or strain type of bacteria. In this study, the tonsil infection with the virulent strains of E. rhusiopathiae (43008) was used as the model. The new perspective that the epithelial cell of swine tonsil was an important infection route for virulent strains of virulent erysipelas would be elucidated for the first time and the mechanism of STAT3-mediated epithelial cell infection further was explored . The contents includes, 1) Establishment of an in vivo infection model to identify epithelial cell infections and then using morphological techniques and laser microdissection to collect infected tonsil epithelial cells. 2) Tonsillar epithelial cells were sorted using immunomagnetic beads technology to establish an in vitro infection model and collect cells. 3) Transcriptome sequencing of epithelial cells collected in vitro and in vivo to deepen the interaction mechanism. 4) Using gene silencing and overexpression techniques to verify STAT3-mediated signaling pathways. This study will reveal the host-pathogen interaction network associated with STAT3 signaling in swine tonsil epithelial cells, providing a theoretical basis for the prevention and control of swine erysipelas disease and the study of new vaccines.

扁桃体黏膜上皮是红斑丹毒丝菌侵入机体的第一道屏障，而黏膜感染途径是否涉及上皮细胞与细菌毒力或毒株类型有关。本研究以红斑丹毒丝菌强毒株(43008)感染扁桃体为模型，首次阐明猪扁桃体上皮细胞是猪丹毒强毒株感染的重要靶细胞，并进一步探索STAT3介导上皮细胞感染的作用机制。研究内容包括：①建立在体感染模型，利用形态学技术确定上皮细胞感染，结合激光显微切割收集感染扁桃体组织上皮细胞；②利用免疫磁珠技术分选扁桃体上皮细胞，建立体外感染模型并收集细胞；③对体内和体外收集的上皮细胞进行转录组测序，深度挖掘基于STAT3的互作机制④利用基因沉默和过表达技术，验证STAT3介导的相关信号途径。本研究将揭示猪扁桃体上皮细胞STAT3信号相关的宿主与病原互作网络，为猪丹毒疫病的防控和新型疫苗的研究提供理论依据。

本项目从红斑丹毒丝菌病原学差异、黏膜感染机制以及实验动物等方面为切入点，深入研究猪扁桃体上皮细胞感染进程，探索STAT3介导上皮细胞感染的作用机制。主要研究内容和获得的主要研究成果：1）建立并优化猪丹毒阴性猪筛选工作程序，为开展猪丹毒相关科学研究和生物制品检验工作提供基础；2）制备了猪丹毒多克隆抗体，为深入研究红斑丹毒丝菌感染机制提供抗体支持；3）首次建立猪丹毒经口感染动物模型，系统研究了感染进程中的时空动态情况，填补了猪丹毒模拟自然感染的动物模型；4）我们发现STAT3是猪扁桃体粘膜上皮感染红斑丹毒丝菌进程中的关键信号通路，为进一步研究猪丹毒黏膜侵染机制提供了数据支持；5）首次建立了小鼠肠黏膜感染猪丹毒模型，揭示了小鼠肠粘膜感染进程中的时空动态分布；6）比较了猪丹毒实验动物敏感性差异研究，为全国猪丹毒疫苗检验检测工作提供重要数据支撑。

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