Vos Joost B, van Sterkenburg Marianne A, Rabe Klaus F, Schalkwijk Joost, Hiemstra Pieter S, Datson Nicole A
Department of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands.
Physiol Genomics. 2005 May 11;21(3):324-36. doi: 10.1152/physiolgenomics.00289.2004. Epub 2005 Feb 8.
The airway epithelium responds to microbial exposure by altering expression of a variety of genes to increase innate host defense. We aimed to delineate the early transcriptional response in human primary bronchial epithelial cells exposed for 6 h to a mixture of IL-1beta and TNF-alpha or heat-inactivated Pseudomonas aeruginosa. Because molecular mechanisms of epithelial innate host defense are not fully understood, the open-ended expression-profiling technique SAGE was applied to construct gene expression profiles covering 30,000 genes: 292 genes were found to be differentially expressed. Expression of seven genes was confirmed by real-time qPCR. Among differentially expressed genes, four classes or families were identified: keratins, proteinase inhibitors, S100 calcium-binding proteins, and IL-1 family members. Marked transcriptional changes were observed for keratins that form a key component of the cytoskeleton in epithelial cells. Expression of antimicrobial proteinase inhibitors SLPI and elafin was elevated after microbial or cytokine exposure. Interestingly, expression of numerous S100 family members was observed, and eight members, including S100A8 and S100A9, were among the most differentially expressed genes. Differential expression was also observed for the IL-1 family members IL-1beta, IL-1 receptor antagonist, and IL-1F9, a newly discovered IL-1 family member. Clustering of differentially expressed genes into biological processes revealed that the early inflammatory response in airway epithelial cells to IL-1beta-TNF-alpha and P. aeruginosa is characterized by expression of genes involved in epithelial barrier formation and host defense.
气道上皮细胞通过改变多种基因的表达来应对微生物暴露，以增强先天性宿主防御。我们旨在描绘人原代支气管上皮细胞在暴露于白细胞介素-1β和肿瘤坏死因子-α混合物或热灭活铜绿假单胞菌6小时后的早期转录反应。由于上皮细胞先天性宿主防御的分子机制尚未完全了解，因此应用开放式表达谱技术SAGE构建覆盖30,000个基因的基因表达谱：发现292个基因存在差异表达。通过实时定量PCR证实了七个基因的表达。在差异表达的基因中，鉴定出四类或四个家族：角蛋白、蛋白酶抑制剂、S100钙结合蛋白和白细胞介素-1家族成员。观察到角蛋白有明显的转录变化，角蛋白是上皮细胞细胞骨架的关键组成部分。微生物或细胞因子暴露后，抗菌蛋白酶抑制剂SLPI和弹性蛋白酶的表达升高。有趣的是，观察到许多S100家族成员的表达，包括S100A8和S100A9在内的八个成员是差异表达最明显的基因之一。白细胞介素-1家族成员白细胞介素-1β、白细胞介素-1受体拮抗剂和新发现的白细胞介素-1家族成员白细胞介素-1F9也存在差异表达。将差异表达的基因聚类到生物学过程中发现，气道上皮细胞对白细胞介素-1β-肿瘤坏死因子-α和铜绿假单胞菌的早期炎症反应的特征是参与上皮屏障形成和宿主防御的基因表达。
