Functional genomics of human bronchial epithelial cells directly interacting with conidia of Aspergillus fumigatus.
Functional genomics of human bronchial epithelial cells directly interacting with conidia of Aspergillus fumigatus.
复制标题
DOI：
10.1186/1471-2164-11-358
复制
发表时间：
2010-06-04
期刊：
影响因子：
4.4
通讯作者：
Tebbutt SJ
中科院分区：
文献类型：
--
作者：
Gomez P;Hackett TL;Moore MM;Knight DA;Tebbutt SJ
烟曲霉（A.烟曲霉）是一种普遍存在的真菌，其通过释放大量的空气传播的分生孢子（孢子）而无性繁殖，所述分生孢子（孢子）易于呼吸。在过敏和免疫功能低下的个体中，A.烟曲霉可引致多种疾病，包括过敏性支气管肺曲霉病，曲霉瘤及侵袭性曲霉病。以往的研究表明，A.烟曲霉分生孢子被巨噬细胞和肺上皮细胞内化;然而，气道上皮细胞对分生孢子的确切转录应答目前尚不清楚。因此，本研究的目的是确定人支气管上皮细胞系（16 HBE 14 o-）与A.烟曲霉分生孢子我们使用荧光激活细胞分选（FACS）来分离具有结合和/或内化的A的16 HBE 140-细胞。表达绿色荧光蛋白的烟曲霉分生孢子来自无孢子的细胞。然后分离总RNA，并使用Agilent全人类基因组微阵列评估16 HBE 140-细胞的转录组。免疫荧光染色和制霉菌素保护测定表明，16 HBE 140-细胞内化30-50%的结合分生孢子内6小时的共孵育。在对相同的细胞培养物进行FAC-sorting以将与分生孢子相关的细胞与没有分生孢子的细胞分开之后，全基因组分析揭示了一组889个基因在具有分生孢子的细胞中显示差异表达。具体地，这些16 HBE 140-细胞具有来自与修复和炎症过程相关的基因的转录物水平增加（例如，基质金属蛋白酶、趋化因子和谷胱甘肽S-转移酶）。此外，差异表达的基因显著富集基因本体论术语，包括：染色质组装，G-蛋白偶联受体结合，趋化因子活性和谷胱甘肽代谢过程（上调）;细胞周期阶段，有丝分裂和细胞内细胞器（下调）。我们展示了一种使用FACs分析来自同一细胞群的感染和未感染细胞的转录组的方法，这将为未来表征病原体（如A.烟曲霉与来自具有或不具有潜在疾病易感性的个体的人细胞。
Aspergillus fumigatus (A. fumigatus) is a ubiquitous fungus which reproduces asexually by releasing abundant airborne conidia (spores), which are easily respirable. In allergic and immunocompromised individuals A. fumigatus can cause a wide spectrum of diseases, including allergic bronchopulmonary aspergillosis, aspergilloma and invasive aspergillosis. Previous studies have demonstrated that A. fumigatus conidia are internalized by macrophages and lung epithelial cells; however the exact transcriptional responses of airway epithelial cells to conidia are currently unknown. Thus, the aim of this study was to determine the transcriptomic response of the human bronchial epithelial cell line (16HBE14o-) following interaction with A. fumigatus conidia. We used fluorescence-activated cell sorting (FACS) to separate 16HBE14o- cells having bound and/or internalized A. fumigatus conidia expressing green fluorescent protein from cells without spores. Total RNA was then isolated and the transcriptome of 16HBE14o- cells was evaluated using Agilent Whole Human Genome microarrays. Immunofluorescent staining and nystatin protection assays demonstrated that 16HBE14o- cells internalized 30-50% of bound conidia within six hrs of co-incubation. After FAC-sorting of the same cell culture to separate cells associated with conidia from those without conidia, genome-wide analysis revealed a set of 889 genes showing differential expression in cells with conidia. Specifically, these 16HBE14o- cells had increased levels of transcripts from genes associated with repair and inflammatory processes (e.g., matrix metalloproteinases, chemokines, and glutathione S-transferase). In addition, the differentially expressed genes were significantly enriched for Gene Ontology terms including: chromatin assembly, G-protein-coupled receptor binding, chemokine activity, and glutathione metabolic process (up-regulated); cell cycle phase, mitosis, and intracellular organelle (down-regulated). We demonstrate a methodology using FACs for analyzing the transcriptome of infected and uninfected cells from the same cell population that will provide a framework for future characterization of the specific interactions between pathogens such as A. fumigatus with human cells derived from individuals with or without underlying disease susceptibility.
登录
查看更多内容
影响因子：
2.9
作者：
Amitani, R.;Kawanami, R.
通讯作者：
Kawanami, R.
DOI：
10.1164/rccm.200603-392oc
发表时间：
2006-11-15
影响因子：
24.7
作者：
Kicic, Anthony;Sutanto, Erika N.;Stick, Stephen M.
通讯作者：
Stick, Stephen M.
影响因子：
3.1
作者：
Ibrahim-Granet, O;Philippe, B;Latgé, JP
通讯作者：
Latgé, JP
影响因子：
3.1
作者：
Cortez, KJ;Lyman, CA;Walsh, TJ
通讯作者：
Walsh, TJ
影响因子：
3
作者：
Bellanger, Anne-Pauline;Millon, Laurence;Bretagne, Stephane
通讯作者：
Bretagne, Stephane