Functional genomics of innate host defense molecules in normal human monocytes in response to Aspergillus fumigatus
Functional genomics of innate host defense molecules in normal human monocytes in response to Aspergillus fumigatus
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DOI：
10.1128/iai.74.4.2353-2365.2006
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发表时间：
2006-04-01
影响因子：
3.1
通讯作者：
Walsh, TJ
中科院分区：
文献类型：
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作者：
Cortez, KJ;Lyman, CA;Walsh, TJ
烟曲霉在感染过程的早期诱导吞噬细胞释放先天免疫相关分子。然而，人们对参与这种反应的多个基因的复杂表达谱知之甚少。因此，我们研究了感染A.利用DNA微阵列技术分析烟曲霉毒素。提取0、2、4和6 h时HMCs的总RNA，线性扩增，杂交到Affytelium HG 133 Plus 2.0基因芯片上，并用Affytelium扫描仪分析。基因表达的变化计算为感染与对照HMC表达的变化的比率。烟曲霉诱导了1,827个基因的差异表达（P < 0.05）。编码细胞因子和趋化因子的基因参与宿主对A.在最初的2至6小时期间，烟曲霉的细胞因子，包括白细胞介素-1 β（IL-1 β）、IL-8、CXCL 2、CCL 4、CCL 3和CCL 20，以及调理素长正五聚蛋白3上调，与吞噬作用的增加一致。同时，编码CD 14、ficolin 1和MARCO的基因下调，编码IL-10和基质金属蛋白酶I的基因上调。编码热休克蛋白40和110以及连接蛋白26和30的基因的上调可能指向新的分子，其在曲霉病发病机制中的作用以前没有报道过。通过逆转录-PCR和酶免疫测定法对所选基因的转录谱进行了验证。因此，A.烟曲霉分生孢子诱导宿主防御和免疫调节中重要基因的协调表达。
Aspergillus fumigatus induces the release of innate immune-related molecules from phagocytic cells early in the course of infection. Little is known, however, about the complex expression profiles of the multiple genes involved in this response. We therefore investigated the kinetics of early gene expression in human monocytes (HMCs) infected with conidia of A. fumigatus using DNA microarray analysis. Total RNA from HMCs at 0, 2, 4, and 6 h was extracted, linearly amplified, hybridized onto Affymetrix HG133 Plus 2.0 gene chips, and analyzed with an Affymetrix scanner. Changes in gene expression were calculated as a ratio of those expressed by infected versus control HMCs. Aspergillus fumigatus induced differential regulation of expression in 1,827 genes (P < 0.05). Genes encoding cytokines and chemokines involved in host defense against A. fumigatus, including interleukin-1 beta (IL-1 beta), IL-8, CXCL2, CCL4, CCL3, and CCL20, as well as the opsonin long pentraxin 3, were up-regulated during the first 2 to 6 h, coinciding with an increase in phagocytosis. Simultaneously, genes encoding CD14, ficolin1, and MARCO were down-regulated, and genes encoding IL-10 and matrix metalloproteinase I were up-regulated. Up-regulation of the genes encoding heat shock proteins 40 and 110 and connexins 26 and 30 may point to novel molecules whose role in the pathogenesis of aspergillosis has not been previously reported. Verification of the transcriptional profiling was obtained for selected genes by reverse transcription-PCR and enzyme immunoassay. Thus, A. fumigatus conidia induced a coordinated expression of genes important in host defense and immunomodulation.