Human corneal epithelial cells produce antimicrobial peptides LL-37 and β-defensins in response to heat-killed Candida albicans.
Human corneal epithelial cells produce antimicrobial peptides LL-37 and β-defensins in response to heat-killed Candida albicans.
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DOI：
10.1159/000357977
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发表时间：
2014
影响因子：
2.1
通讯作者：
Li DQ
中科院分区：
文献类型：
--
作者：
Hua X;Yuan X;Tang X;Li Z;Pflugfelder SC;Li DQ
目的探讨人角膜上皮细胞（HCECs）对真菌的天然反应，并通过产生抗微生物肽LL-37和β-防御素（β-defensins）进行初步研究。用不同剂量（10 3 ~ 10 6 cells/ml）的热灭活白念珠菌（HKCA）处理原代HCEC 2-48 h。对细胞进行总RNA提取、逆转录和mRNA表达的定量实时PCR。将处理48 h的细胞用于免疫荧光染色和ELISA。在正常HCEC中检测到人LL-37和β-防御素（hBD）1-4。HKCA可剂量依赖性地诱导LL-37、hBD 2和hBD 3的mRNA表达，其峰值出现在4 h。HKCA（106 cells/ml）对HCECs中LL-37、hBD 2和hBD 3的mRNA的刺激作用分别为4.33 ± 1.81、3.75 ± 1.31和4.91 ± 1.09倍。通过免疫荧光染色和ELISA在蛋白质水平证实HKCA刺激LL-37、hBD 2和hBD 3的产生。与未处理的HCEC相比，在HKCA（106个细胞/ml）处理的HCEC的培养基中，LL-37、hBD 2和hBD 3的蛋白质产量分别显著增加至109.1 ± 18.2pg/ml、4.33 ± 1.67ng/ ml和296.9 ± 81.8pg/ml。HCECs在HKCA的刺激下产生抗微生物肽LL-37、hBD 2和hBD 3，这表明眼表防御真菌侵袭的一种新的先天免疫机制。
To explore the innate response of human corneal epithelial cells (HCECs) exposed to fungus by producing antimicrobial peptides LL-37 and β-defensins. Primary HCECs were treated with heat-killed Candida albicans (HKCA) at different doses (10 3 –10 6 cells/ml) for 2–48 h. The cells were subjected to total RNA extraction, reverse transcription and quantitative real-time PCR for mRNA expression. Cells treated for 48 h were used for immunofluorescent staining and ELISA. Human LL-37 and β-defensins (hBDs) 1–4 were detected in normal HCECs. The mRNA expression of LL-37, hBD2, and hBD3 was dose-dependently induced by HKCA with their peak levels at 4 h. HKCA (106 cells/ml) stimulated the mRNA of LL-37, hBD2, and hBD3 4.33 ± 1.81, 3.75 ± 1.31, and 4.91 ± 1.09 fold, respectively, in HCECs. The stimulated production of LL-37, hBD2, and hBD3 by HKCA was confirmed at protein levels by immunofluorescent staining and ELISA. The protein production of LL-37, hBD2, and hBD3 significantly increased to 109.1 ± 18.2 pg/ml, 4.33 ± 1.67 ng/ ml, and 296.9 ± 81.8 pg/ml, respectively, in culture medium of HCECs exposed to HKCA (10 6 cells/ml) compared to untreated HCECs. HCECs produce antimicrobial peptides, LL-37, hBD2 and hBD3, in response to stimulation of HKCA, which suggests a novel innate immune mechanism of the ocular surface in defense against fungal invasion.
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