School of Optometry and Ophthalmology, Wenzhou Medical College, Wenzhou, China; Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX, USA.
School of Optometry and Ophthalmology, Wenzhou Medical College, Wenzhou, China.
Exp Eye Res. 2014 Mar;120:118-26. doi: 10.1016/j.exer.2014.01.014. Epub 2014 Jan 29.
The association and mechanism of bacteria linking to the allergic inflammation have not been well elucidated. This study was to explore a potential link between bacterial pathogens and allergic conjunctivitis by dendritic cells (DCs). Bone marrow-derived DCs from BALB/c and MyD88 knockout mice were treated with or without bacterial pathogens or thymic stromal lymphopoietin (TSLP). Two murine models of the topical challenge with LPS or flagellin and experimental allergic conjunctivitis (EAC) were used for in vivo study. The mRNA expression was determined by reverse transcription and real time PCR, and protein production was evaluated by ELISA, Western blotting, immunofluorescent staining and flow cytometry. TSLP mRNA and protein were found to be largely induced by DCs challenged with microbial pathogens, highly by lipopolysaccharide (LPS) and flagellin. The expression of MyD88, NFκB1, NFκB2 and RelA accompanied by NFκB p65 nuclear translocation and TSLP induction were significantly stimulated by flagellin, but blocked by TLR5 antibody or NFκB inhibitor in DCs from MyD88(+/+) but not MyD88(-/-) mice. TSLP promoted the expression of CD40, CD80, OX40 ligand (OX40L), IL-13 and CCL17 by DCs. TSLP-producing DCs were identified in vivo in ocular surface conjunctiva and draining cervical lymph nodes from two murine models of topical challenge with LPS or flagellin, and EAC in BALB/c mice. TSLP/TSLPR/OX40L signaling was observed in DCs of EAC mice. Our findings demonstrate that DCs not only respond to TSLP, but also produce TSLP via TLR/MyD88/NFκB pathways in response to bacterial pathogens, suggesting a potential link between bacteria and allergic disease.
细菌与过敏炎症的关联和机制尚未得到充分阐明。本研究旨在通过树突状细胞（DC）探索细菌病原体与过敏性结膜炎之间的潜在联系。来自 BALB/c 和 MyD88 敲除小鼠的骨髓来源的 DC 用或不用细菌病原体或胸腺基质淋巴细胞生成素（TSLP）处理。使用 LPS 或鞭毛蛋白局部挑战的两种小鼠模型和实验性过敏性结膜炎（EAC）进行体内研究。通过逆转录和实时 PCR 确定 mRNA 表达，通过 ELISA、Western blot、免疫荧光染色和流式细胞术评估蛋白产生。发现 TSLP mRNA 和蛋白主要由受微生物病原体挑战的 DC 诱导，尤其是脂多糖（LPS）和鞭毛蛋白。MyD88、NFκB1、NFκB2 和 RelA 的表达以及 NFκB p65 核易位和 TSLP 诱导伴随着 NFκB 受体激活剂（NFκB1）的表达，由鞭毛蛋白显著刺激，但在 MyD88(+/+)而不是 MyD88(-/-)小鼠的 DC 中被 TLR5 抗体或 NFκB 抑制剂阻断。TSLP 促进 DC 表达 CD40、CD80、OX40 配体（OX40L）、IL-13 和 CCL17。在 LPS 或鞭毛蛋白局部挑战的两种小鼠模型以及 BALB/c 小鼠的 EAC 中，在眼表面结膜和引流颈淋巴结中鉴定出产生 TSLP 的 DC。在 EAC 小鼠的 DC 中观察到 TSLP/TSLPR/OX40L 信号。我们的研究结果表明，DC 不仅对 TSLP 有反应，而且还通过 TLR/MyD88/NFκB 途径对细菌病原体产生 TSLP，提示细菌与过敏性疾病之间存在潜在联系。
