过敏性哮喘患者过敏原吸入激发后外周血基因表达变化
2012/04/06
目的：（1）研究珠蛋白mRNA去除在检测外周血基因差异性表达中的作用；（2）研究过敏性哮喘患者过敏原激发后外周血基因表达变化。
方法：入选9名20～60岁患有稳定、轻度过敏性哮喘患者，并进行过敏原吸入激发试验。所有患者均出现早期的哮喘反应，表现为第一秒用力呼气体积下降≥20%。在激发试验前即刻和试验后2小时，采用PAXgene试管（n=4）、EDTA试管（n=5）收集血液。PAXgene试管分装的样品进行珠蛋白还原实验（PAX-GR）。采用Affymetrix GeneChip(®) Human Gene 1.0 ST array进行转录组分析。采用基因芯片总结因素分析对数据进行处理，采用基因芯片线性模型对数据进行分析。采用精巧旁路分析对旁路进行分析。
结果：珠蛋白还原实验发现了一些低丰度的探针。然而，与无珠蛋白的PAXgene样本（PAX-NGR）相比，珠蛋白还原实验显著降低了检测差异性表达基因（DEGs）的能力。4对PAX-NGR和5对EDTA样本的联合转录分析，找到1595个与过敏原吸入激发相关的DEG（假阳性率≤ 5%），这些DEG反应主要表现为炎症反应、细胞迁移和免疫细胞转运。
结论：我们结果显示，进行珠蛋白还原后，检测DEG的能力显著下降。此外，对于轻度哮喘患者，在过敏原吸入激发试验2小时后，外周血转录组存在显著的变化。
（林江涛 审校）
J Asthma. 2012 Feb 9. [Epub ahead of print]
Peripheral Blood Gene Expression Changes during Allergen Inhalation Challenge in Atopic Asthmatic Individuals.
Kam SH, Singh A, He JQ, Ruan J, Gauvreau GM, O’Byrne PM, Fitzgerald JM, Tebbutt SJ.
Source
James Hogg Research Centre, St. Paul’s Hospital, University of British Columbia , Vancouver, BC , Canada.
Abstract
OBJECTIVES：(1) To investigate the effects of globin mRNA depletion in detecting differential gene expression in peripheral blood and (2) to investigate changes in peripheral blood gene expression in atopic asthmatic individuals undergoing allergen inhalation challenge.
METHODS：Asthmatic subjects (20-60 years of age, with stable, mild allergic asthma, n = 9) underwent allergen inhalation challenges. All had an early asthmatic response of ≥20% fall in forced expiratory volume in 1 second. Blood was collected immediately prior to and 2 hours after allergen challenge using PAXgene tubes (n = 4) and EDTA tubes (n = 5). Aliquots of the PAXgene blood samples were subjected to globin reduction (PAX-GR). Transcriptome analysis was performed using Affymetrix GeneChip(®) Human Gene 1.0 ST arrays. Data were preprocessed using factor analysis for robust microarray summarization and analyzed using linear models for microarrays. Pathway analyses were performed using Ingenuity Pathway Analysis.
RESULTS：Globin reduction uncovered probe sets of lower abundance. However, it significantly reduced the ability to detect differentially expressed genes (DEGs) when compared to non-globin-reduced PAXgene samples (PAX-NGR). Combined transcriptional analysis of four PAX-NGR and five EDTA sample pairs identified 1595 DEGs associated with allergen inhalation challenge (false discovery rate ≤ 5%), with the top-ranked network of perturbed biological functions consisting of inflammatory response, cellular movement, and immune cell trafficking.
CONCLUSIONS：While we have demonstrated a diminished ability to detect DEGs after globin reduction, we have nevertheless identified significant changes in the peripheral blood transcriptome of people with mild asthma 2 hours after allergen inhalation challenge.
J Asthma. 2012 Feb 9. [Epub ahead of print]
