Nucleotide oligomerization domain 2 contributes to the innate immune response in THCE cells stimulated by Aspergillus fumigatus conidia
Nucleotide oligomerization domain 2 contributes to the innate immune response in THCE cells stimulated by Aspergillus fumigatus conidia
复制标题
核苷酸寡聚化结构域 2 有助于烟曲霉分生孢子刺激的 THCE 细胞中的先天免疫反应。
DOI：
10.3980/j.issn.2222-3959.2012.04.01
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发表时间：
2012-08-18
影响因子：
1.4
通讯作者：
Xu, Qiang
中科院分区：
文献类型：
--
作者：
Xu, Zheng-Jie;Zhao, Gui-Qiu;Xu, Qiang
目的：研究核苷酸寡聚结构域2 （NOD2）在永生化人角膜上皮细胞系（THCE）中的表达及其在灭活烟曲霉（Aspergillus fumigatus, Af）分生孢子引发的先天免疫应答中的作用。方法：以正常THCE细胞为对照。与失活的分生孢子孵育0.5、2、4、6和8小时后，收集THCE细胞，RT-PCR检测NOD2和受体相互作用蛋白2 (RIP2) mRNA表达量。ELISA法分析细胞上清液中细胞内NOD2、nf - κ B蛋白及促炎因子tnf - α、IL-8、IL-6的表达。结果：我们的数据表明，NOD2在正常THCE细胞中表达。被失活的分生孢子触发后，NOD2、RIP2 mRNA的表达以及NOD2、NF-kappa B、tnf - α、IL-8、IL-6的分泌均呈时间依赖性增加，分别在4、6、6、4、6、6、4小时达到峰值。经NOD2中和抗体预处理后，RIP2、NF-kappa B、tnf - α、IL-8的表达均在峰值处显著下降，而IL-6的分泌变化不大。结论：本研究结果提示，NOD2在THCE细胞中存在并表达，并通过nf - κ B途径诱导tnf - α、IL-8等促炎细胞因子参与失活afcon孢子引发的先天免疫应答。
AIM: To investigate the expression of nucleotide oligomerization domain 2 (NOD2) in the immortalized human corneal epithelial cell line (THCE), and its role in the innate immune response triggered by inactive Aspergillus fumigatus (Af) conidia.METHODS: The normal THCE cells were investigated as controls. After incubation with inactive Af conidia for 0.5, 2, 4, 6, and 8 hours, THCE cells were harvested, mRNA expression of NOD2 and receptor interacting protein 2 (RIP2) was detected by RT-PCR. Intracellular proteins including NOD2, NF-kappa B and proinflammatory cytokines such as TNF-alpha, IL-8, IL-6 in the cell supernatant were analyzed by ELISA.RESULTS: Our data indicate that NOD2 expressed in the normal THCE cells. After triggered by the inactive Af conidia, the expression of NOD2, RIP2 mRNA and the secretion of NOD2, NF-kappa B, TNF-alpha, IL-8, IL-6 both increased in a time-depended manner, and reached the peak point at 4, 6, 6, 4, 6, 6, 4 hours, respectively. And after pretreated with NOD2 neutralizing antibody, the expression of RIP2, NF-kappa B, TNF-alpha, IL-8 both decreased dramatically at the peak point, while the secretion of IL-6 changed little.CONCLUSION: The results of this study suggest that NOD2 exists and expresses in the THCE cells, and contributes to the innate immune responses triggered by inactive Afconidia by induction of proinflammatory cytokines such as TNF-alpha and IL-8 through the NF-kappa B pathway.