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Abstract：
OBJECTIVE: Oral epithelial cells serve as the primary defense against microbial exposure in the oral cavity, including the fungus Candida albicans. Dectin-1 is crucial for recognition of β-glucan in fungi. However, expression and function of Dectin-1 in oral epithelial cells remain unclear.
METHODS: We assessed Dectin-1 expression in Ca9-22 (gingiva), HSC-2 (mouth), HSC-3 (tongue), and HSC-4 (tongue) human oral epithelial cells using flow cytometry and real-time polymerase chain reaction. Cell treated with β-glucan-rich zymosan were evaluated using real-time polymerase chain reaction. Phosphorylation of spleen-associated tyrosine kinase (SYK) was analyzed by western blotting.
RESULTS: Dectin-1 was expressed in all four cell types, with high expression in Ca9-22 and HSC-2. In Ca9-22 cells, exposure to β-glucan-rich zymosan did not alter the mRNA expression of chemokines nor of interleukin (IL)6, IL8, IL1β, IL17A, and IL17F. Zymosan induced the expression of antimicrobial peptides β-defensin-1 and LL-37, but not S100 calcium-binding protein A8 (S100A8) and S100A9. Furthermore, the expression of cylindromatosis (CYLD), a negative regulator of nuclear factor kappa B (NF-κB) signaling, was induced. In HSC-2 cells, zymosan induced the expression of IL17A. The expression of tumor necrosis factor alpha-induced protein 3 (TNFAIP3), a negative regulator of NF-κB signaling, was also induced. Expression of other cytokines and antimicrobial peptides remained unchanged. Zymosan induced phosphorylation of SYK in Ca9-22 cells, as well as NF-κB.
CONCLUSIONS: Oral epithelial cells express Dectin-1 and recognize β-glucan, which activates SYK and induces the expression of antimicrobial peptides and negative regulators of NF-κB, potentially maintaining oral homeostasis.
METHODS: We assessed Dectin-1 expression in Ca9-22 (gingiva), HSC-2 (mouth), HSC-3 (tongue), and HSC-4 (tongue) human oral epithelial cells using flow cytometry and real-time polymerase chain reaction. Cell treated with β-glucan-rich zymosan were evaluated using real-time polymerase chain reaction. Phosphorylation of spleen-associated tyrosine kinase (SYK) was analyzed by western blotting.
RESULTS: Dectin-1 was expressed in all four cell types, with high expression in Ca9-22 and HSC-2. In Ca9-22 cells, exposure to β-glucan-rich zymosan did not alter the mRNA expression of chemokines nor of interleukin (IL)6, IL8, IL1β, IL17A, and IL17F. Zymosan induced the expression of antimicrobial peptides β-defensin-1 and LL-37, but not S100 calcium-binding protein A8 (S100A8) and S100A9. Furthermore, the expression of cylindromatosis (CYLD), a negative regulator of nuclear factor kappa B (NF-κB) signaling, was induced. In HSC-2 cells, zymosan induced the expression of IL17A. The expression of tumor necrosis factor alpha-induced protein 3 (TNFAIP3), a negative regulator of NF-κB signaling, was also induced. Expression of other cytokines and antimicrobial peptides remained unchanged. Zymosan induced phosphorylation of SYK in Ca9-22 cells, as well as NF-κB.
CONCLUSIONS: Oral epithelial cells express Dectin-1 and recognize β-glucan, which activates SYK and induces the expression of antimicrobial peptides and negative regulators of NF-κB, potentially maintaining oral homeostasis.
摘要：
目的：口腔上皮细胞是口腔微生物暴露的主要防御，包括真菌白色念珠菌.Dectin-1对于识别真菌中的β-葡聚糖至关重要。然而,Dectin-1在口腔上皮细胞中的表达和功能尚不清楚。
方法：我们评估了Ca9-22（牙龈）中Dectin-1的表达，HSC-2(口),HSC-3（舌），和HSC-4（舌）人口腔上皮细胞使用流式细胞术和实时聚合酶链反应。使用实时聚合酶链反应评估用富含β-葡聚糖的酵母聚糖处理的细胞。通过蛋白质印迹分析脾相关酪氨酸激酶(SYK)的磷酸化。
结果：Dectin-1在所有四种细胞类型中均有表达，在Ca9-22和HSC-2中具有高表达。在Ca9-22细胞中,暴露于富含β-葡聚糖的酵母聚糖不会改变趋化因子或白细胞介素（IL）6，IL8，IL1β的mRNA表达，IL17A,IL17F酵母聚糖诱导抗菌肽β-防御素-1和LL-37的表达，但不诱导S100钙结合蛋白A8（S100A8）和S100A9的表达。此外，圆柱瘤病（CYLD）的表达，核因子κB（NF-κB）信号的负调节因子，是诱导的。在HSC-2细胞中，酵母聚糖诱导IL17A的表达。肿瘤坏死因子α诱导蛋白3(TNFAIP3)的表达,NF-κB信号的负调节因子，也被诱导了。其他细胞因子和抗菌肽的表达保持不变。酵母聚糖诱导Ca9-22细胞中SYK的磷酸化,以及NF-κB。
结论：口腔上皮细胞表达Dectin-1并识别β-葡聚糖，激活SYK并诱导抗菌肽和NF-κB负调节因子的表达，可能维持口腔稳态。
方法：我们评估了Ca9-22（牙龈）中Dectin-1的表达，HSC-2(口),HSC-3（舌），和HSC-4（舌）人口腔上皮细胞使用流式细胞术和实时聚合酶链反应。使用实时聚合酶链反应评估用富含β-葡聚糖的酵母聚糖处理的细胞。通过蛋白质印迹分析脾相关酪氨酸激酶(SYK)的磷酸化。
结果：Dectin-1在所有四种细胞类型中均有表达，在Ca9-22和HSC-2中具有高表达。在Ca9-22细胞中,暴露于富含β-葡聚糖的酵母聚糖不会改变趋化因子或白细胞介素（IL）6，IL8，IL1β的mRNA表达，IL17A,IL17F酵母聚糖诱导抗菌肽β-防御素-1和LL-37的表达，但不诱导S100钙结合蛋白A8（S100A8）和S100A9的表达。此外，圆柱瘤病（CYLD）的表达，核因子κB（NF-κB）信号的负调节因子，是诱导的。在HSC-2细胞中，酵母聚糖诱导IL17A的表达。肿瘤坏死因子α诱导蛋白3(TNFAIP3)的表达,NF-κB信号的负调节因子，也被诱导了。其他细胞因子和抗菌肽的表达保持不变。酵母聚糖诱导Ca9-22细胞中SYK的磷酸化,以及NF-κB。
结论：口腔上皮细胞表达Dectin-1并识别β-葡聚糖，激活SYK并诱导抗菌肽和NF-κB负调节因子的表达，可能维持口腔稳态。
