Role of chemokine CCL20 and its receptor CCR6 against bacterial infection.

本研究旨在探讨趋化因子CCL20-CCR6系统在感染沙门菌和其他微生物的小鼠肠道和淋巴器官中的作用。我们使用无免疫小鼠和经口免疫鼠伤寒沙门菌的小鼠，分别分析了鼠伤寒沙门菌毒力菌株攻毒后的一次免疫反应和二次免疫反应。利用免疫后的亚美尼亚仓鼠脾细胞，建立抗小鼠CCR6单克隆抗体29.2L17。该单克隆抗体29.2L17在18种小鼠趋化因子受体中对CCR6具有特异性，能够染色已知表达CCR6的正常小鼠淋巴样细胞。此外，29.2L17可用于免疫组化染色小鼠组织中表达CCR6的细胞。然而，29.2L17对小鼠CCR6没有明显的中和活性。建立鼠伤寒沙门菌经口免疫和经口攻毒的实验条件。在此条件下，分别于鼠伤寒沙门菌攻毒前后的不同时间点采集对照组或免疫小鼠的肠道、肝脏和脾脏。这些组织可用于分析CCL20-CCR6系统在粘膜免疫中原发性和继发性免疫反应中的作用。我们还证明了角膜上皮细胞和间质角质细胞在体外和体内产生CCL20的能力。通过小鼠疱疹性间质角膜炎模型，我们发现除了上皮细胞外，间质角膜炎细胞也能在角膜中有效地产生CCL20，并将表达CCR6的细胞如树突状细胞募集到发炎的角膜中。

This research is to investigate the role of the chemokine CCL20-CCR6 system at intestine and lymphoid organ in mice infected with Salmonella and other microorganisms We used non-immune mice and mice immunized perorally with avirulent strain of Salmonella typhimurium for the analysis of primary immune response and secondary immune response, respectively, after challenge with the virulent strain of Salmonella typhimurium.Using immunized Armenian hamster spleen cells, we established anti-mouse CCR6 monoclonal antibody termed 29.2L17. The monoclonal antibody 29.2L17 was specific for CCR6 among the 18 mouse chemokine receptors and capable of staining normal mouse lymphoid cells known to expree CCR6. Furthermore, 29.2L17 could be used for staining cells expressing CCR6 in mouse tissues by immunohistochemistry. However, 29.2L17 does not have a significant neutralizing activity against mouse CCR6.We established experimental conditions for peroral immunization and peroral challenge of Salmonella typhimurium. Using these conditions, intestinal tract, liver and spleen were collected from control or immunized mice before or after the challenge with Salmonella typhimurium at several time points. These tissues are useful for the analysis of the role of the CCL20-CCR6 system in primary and secondary immune responses in the mucosal immunity.We have also demonstrated the ability of corneal epithelial cells and stromal keratocytes to produce CCL20 in vitro and in vivo. By using a mouse model of herpetic stromal keratatis, it has been revealed that not only epithelial cells but also stromal keratocytes efficiently produce CCL20 in the cornea and recruit cells expressing CCR6 such as dendritic sells into inflamed cornea.