Objective To investigate the effects of Trichophyton rubrum exposure on the expressions of toll-like receptor-2 (TLR-2), TLR-4 and dendritic cell associated C-type lectin-1 (Dectin-1) and cytokine secretions in human keratinocytes cell line HaCaT. Methods The Mrna of TLR-2,4, and dectin-1 in the HaCaT co-cultured with the conidia of Trichophyton rubrum conidia for 24 h was measured, with real-time PCR. The mean fluorescence intensity and the percentage of cells positive for TLR-2, 4, and dectin-1 was detected during the co-culture using flow cytometry. The cytokine secretion profiles in the cell culture supernatant was analyzed using a cytokine antibody array. Results The TLR-2,4, and dectin-1 Mrna expressions, mean fluorescence intensity and percentage of positive cells for TLR-2,4, and dectin-1 all increased in HaCaT cells in response to Ttrichophyton rubrum conidia exposure. The results of cytokine antibody array demonstrated obviously increased secretions of IL-8, 1-309, IFN-7, IL-6, and IL-13 in the culture supernatant of HaCaT cells in response to Ttrichophyton rubrum exposure. Conclusion The immune responses and immunological recognition of human keratinocytes to Trichophyton rubrum conidia are partially mediated by up-regulating the expressions of TLR-2, TLR-4 and dectin-1 and secretions of multiple cytokines.%目的 研究红色毛癣菌对人角质形成细胞模式识别受体TLR-2,TLR-4,Dectin-1表达及细胞因子分泌的影响,探讨角质形成细胞对红色毛癣菌的免疫应答及其机制.方法 红色毛癣菌孢子与人永生化表皮细胞株HaCaT细胞共培养,采用Real time PCR检测共培养后HaCaT细胞TLR-2,TLR-4,Dectin-lmRNA的表达情况;采用流式细胞技术检测共培养后不同时间段HaCaT细胞TLR-2,TLR-4及Dectin-1平均荧光强度;采用蛋白芯片抗体阵列检测共培养上清液中36种不同的细胞因子,趋化因子和急性时相蛋白的表达情况.结果 共培养6 h后,TLR-2,TLR-4,Dectin-lmRNA表达上调;共培养24 h后TLR-2,TLR-4,Dectin-1平均荧光强度增高,细胞因子IL-8,I-309,IFN-γ,IL-6,IL-13在红色毛癣菌刺激后分泌量明显增加.结论 人角质形成细胞对红色毛癣菌的免疫识别和应答,在一定程度上可通过上调HaCaT细胞中模式识别受体TLR-2,TLR-4及Dectin-1后分泌多种细胞因子来实现.
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