Poly(I:C)激活的鼻成纤维细胞促进CRSwNP中嗜酸性粒细胞的募集

文献信息

中文摘要

背景： 慢性鼻窦炎伴鼻息肉（CRSwNP）以持续性黏膜炎症和嗜酸性粒细胞浸润为特征。尽管上皮细胞和免疫细胞已被广泛研究，但鼻成纤维细胞在固有免疫激活和嗜酸性粒细胞募集中的作用仍不清楚。
方法： 分析来自鼻窦组织的公共单细胞RNA测序数据，以确定TLR3和炎症介质的细胞分布。在患者来源的鼻窦组织中验证TLR3表达。用聚肌胞苷酸（Poly(I:C)）刺激原代人鼻成纤维细胞，并通过实时PCR和ELISA评估细胞因子和趋化因子的表达。使用药理学抑制剂和蛋白质印迹法检测信号通路。使用微流控趋化平台评估嗜酸性粒细胞向Poly(I:C)刺激的成纤维细胞的迁移。
结果： TLR3和炎症介质（包括CCL2、IL6、CXCL8和CCL11）优先富集于基质细胞中。成纤维细胞亚聚类显示，CRSwNP中活化的、重塑的和炎症性成纤维细胞状态扩增，并伴有TLR3和趋化因子表达增加。CRSwNP中组织TLR3表达增加，并与Lund-Mackay CT评分呈正相关。Poly(I:C)刺激在原代鼻成纤维细胞中诱导IL-6、CXCL8、CCL2和CCL11的剂量依赖性产生。这些反应通过TRIF、p38、JNK和PI3K/AKT信号传导介导。在功能上，Poly(I:C)刺激的成纤维细胞促进嗜酸性粒细胞趋化，而抑制TRIF、JNK和PI3K信号传导可减弱这种趋化。
结论： 鼻成纤维细胞在CRSwNP中作为TLR3依赖性固有免疫激活的活跃基质介质发挥作用。成纤维细胞来源的炎症介质和嗜酸性粒细胞募集介质可能将病毒样刺激与CRSwNP中的嗜酸性炎症和组织重塑联系起来。

英文摘要

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by persistent mucosal inflammation and eosinophilic infiltration. Although epithelial and immune cells have been widely studied, the role of nasal fibroblasts in innate immune activation and eosinophil recruitment remains unclear.
METHODS: Public single-cell RNA sequencing data from sinonasal tissues were analyzed to define the cellular distribution of TLR3 and inflammatory mediators. TLR3 expression was validated in patient-derived sinonasal tissues. Primary human nasal fibroblasts were stimulated with polyinosinic-polycytidylic acid (Poly(I:C)), and cytokine and chemokine expression was assessed by real-time PCR and ELISA. Signaling pathways were examined using pharmacologic inhibitors and western blotting. Eosinophil migration toward Poly(I:C)-stimulated fibroblasts was evaluated using a microfluidic chemotaxis platform.
RESULTS: TLR3 and inflammatory mediators, including CCL2, IL6, CXCL8, and CCL11, were preferentially enriched in stromal cells. Fibroblast subclustering revealed expansion of activated, remodeling, and inflammatory fibroblast states in CRSwNP, accompanied by increased TLR3 and chemokine expression. Tissue TLR3 expression was increased in CRSwNP and positively correlated with Lund-Mackay CT scores. Poly(I:C) stimulation induced dose-dependent production of IL-6, CXCL8, CCL2, and CCL11 in primary nasal fibroblasts. These responses were mediated through TRIF, p38, JNK, and PI3K/AKT signaling. Functionally, Poly(I:C)-stimulated fibroblasts promoted eosinophil chemotaxis, which was attenuated by inhibition of TRIF, JNK, and PI3K signaling.
CONCLUSIONS: Nasal fibroblasts function as active stromal mediators of TLR3-dependent innate immune activation in CRSwNP. Fibroblast-derived inflammatory and eosinophil-recruiting mediators may link viral-like stimulation to eosinophilic inflammation and tissue remodeling in CRSwNP.