评价鼠肺上皮细胞（TC-1 JHU-1）系对烟曲霉的反应中 Th2 促进细胞因子 IL-25/IL-33/TSLP 和基因 TLR2/TLR4 的产生。
Evaluation of murine lung epithelial cells (TC-1 JHU-1) line to develop Th2-promoting cytokines IL-25/IL-33/TSLP and genes Tlr2/Tlr4 in response to Aspergillus fumigatus.
机构信息
Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
Department of Pathobiology, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Amol, Iran.
出版信息
J Mycol Med. 2018 Jun;28(2):349-354. doi: 10.1016/j.mycmed.2018.02.001. Epub 2018 Mar 7.
OBJECTIVE
The aims of this study were to determine the role of live and heat-killed Aspergillus fumigatus conidia in releasing interleukin (IL)-25, IL-33 and thymic stromal lymphopoietin (TSLP) and to express Toll-like receptor (Tlr)2 and Tlr4 genes.
MATERIALS AND METHODS
Murine lung epithelial cells were incubated with live and heat-killed A. fumigatus conidia at 37°C for 6, 24 and 48h. After treatments, ELISA was performed to measure the concentrations of IL-25, IL-33 and TSLP in the supernatants. Quantitative real-time PCR (qPCR) was performed to assess the expression levels of Tlr2 and Tlr4 genes.
RESULTS
The concentrations of IL-25 and IL-33 significantly increased after exposure to live and heat-killed conidia for various times when compared with untreated control (P<0.05). The secretion of TSLP at different concentrations of heat-killed conidia was significantly higher than both live conidia and untreated control (P<0.05). qRT-PCR results indicated a up-regulation from 1.08 to 3.60-fold for Tlr2 gene expression and 1.20 to 1.80-fold for Tlr4 gene expression exposed to heat-killed conidia.
CONCLUSION
A. fumigatus has a potential ability to stimulate murine lung epithelial cells to produce IL-25/IL-33/TSLP, as well as to express Tlr2/Tlr4 genes, indicating an important role of lung epithelial cells in innate immune responses to A. fumigatus interaction.
目的
本研究旨在确定活的和热灭活的烟曲霉分生孢子在释放白细胞介素（IL）-25、IL-33 和胸腺基质淋巴细胞生成素（TSLP）中的作用，并表达 Toll 样受体（Tlr）2 和 Tlr4 基因。
材料和方法
将鼠肺上皮细胞在 37°C 下与活的和热灭活的 A. fumigatus 分生孢子孵育 6、24 和 48 小时。处理后，通过 ELISA 法测量上清液中 IL-25、IL-33 和 TSLP 的浓度。通过实时定量 PCR（qPCR）评估 Tlr2 和 Tlr4 基因的表达水平。
结果
与未处理的对照组相比，暴露于活的和热灭活的分生孢子后，IL-25 和 IL-33 的浓度在不同时间点均显著增加（P<0.05）。不同浓度热灭活分生孢子的 TSLP 分泌量明显高于活分生孢子和未处理对照组（P<0.05）。qRT-PCR 结果表明，与活分生孢子和未处理对照组相比，Tlr2 基因表达上调 1.08 至 3.60 倍，Tlr4 基因表达上调 1.20 至 1.80 倍。
结论
A. fumigatus 具有刺激鼠肺上皮细胞产生 IL-25/IL-33/TSLP 的潜在能力，以及表达 Tlr2/Tlr4 基因的能力，表明肺上皮细胞在对 A. fumigatus 相互作用的固有免疫反应中具有重要作用。
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