Analysis by high density cDNA arrays of altered gene expression in human intestinal epithelial cells in response to infection with the invasive enteric bacteria Salmonella
Analysis by high density cDNA arrays of altered gene expression in human intestinal epithelial cells in response to infection with the invasive enteric bacteria Salmonella
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DOI：
10.1074/jbc.275.19.14084
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发表时间：
2000-05-12
影响因子：
4.8
通讯作者：
Kagnoff, MF
中科院分区：
文献类型：
--
作者：
Eckmann, L;Smith, JR;Kagnoff, MF
许多临床上重要的肠道：病原体通过侵入和穿过肠上皮而引发疾病，这一过程伴随着促炎细胞因子在肠上皮中表达的增加。为了进一步明确肠上皮细胞在启动和调节宿主对侵袭性细菌感染的反应中所起的作用，采用高密度基因芯片杂交选择的方法，研究了人肠上皮细胞感染典型侵袭性细菌沙门氏菌后，类似于4300个基因的mRNA表达谱。通过逆转录-聚合酶链式反应、Northern印迹分析和蛋白质分析对选定的结果进行进一步评估。上皮性感染沙门氏菌后，在检测的所有基因中，有相对较小一部分的mRNA表达显着上调。其中以粒细胞集落刺激因子、抑制素A、EB病毒诱导基因3、白细胞介素8、巨噬细胞炎性蛋白-2α等细胞因子、TKT、ECK、HEK、转录因子(干扰素调节因子-1)和人类白细胞抗原I类(HLAI)最为显著。此外，转录因子核因子-kappa B被证明是重要的诱导性的mRNA表达的广泛的一组基因测试。这些发现扩大了已知的上皮细胞对侵袭性肠道病原体感染的反应。研究结果还表明，通过基因芯片分析来评估基因表达谱是描述和理解寄主-病原体相互作用的一种有效方法。
Many clinically important enteric: pathogens initiate disease by invading and passing through the intestinal epithelium, a process accompanied by increased epithelial expression of proinflammatory cytokines. To further define the role intestinal epithelial cells play in initiating and modulating the host response to infection with invasive bacteria, hybrid selection on high density cDNA arrays was used to characterize the mRNA expression profile of similar to 4,300 genes in human intestinal epithelial cells after infection with the prototypic invasive bacteria, Salmonella. Selected findings were further evaluated by reverse transcription-polymerase chain reaction, Northern blot analysis, and protein assays. Epithelial infection with Salmonella significantly up-regulated mRNA expression of a relatively small fraction of all genes tested. Of these, several cytokines (granulocyte colony-stimulating factor, inhibin A, Epstein-Barr virus-induced gene 3, interleukin-8, macrophage inflammatory protein-2 alpha), kinases (TKT, Eck, HEK), transcription factors (interferon regulatory factor-1), and HLA class I were the most prominent. Furthermore, the transcription factor NF-kappa B is shown to be important for inducible mRNA expression for a broad group of genes tested. These findings expand the repertoire of known epithelial cell responses to infection with an invasive enteric pathogen. The results also show that evaluation of mRNA expression profiles by cDNA array analysis is a powerful approach to characterizing and understanding host-pathogen interactions.