Development of an inhaled endotoxin challenge protocol for characterizing evoked cell surface phenotype and genomic responses of airway cells in allergic individuals
Development of an inhaled endotoxin challenge protocol for characterizing evoked cell surface phenotype and genomic responses of airway cells in allergic individuals
复制标题
DOI：
10.1016/s1081-1206(10)60444-9
复制
发表时间：
2008-03-01
影响因子：
5.9
通讯作者：
Peden, David B.
中科院分区：
文献类型：
--
作者：
Alexis, Neil E.;Brickey, Willie June;Peden, David B.
背景：环境暴露于内毒素是哮喘恶化的已知原因。吸入内毒素方案已用于评估健康志愿者中与抗原呈递和先天免疫相关的气道细胞表面表型，但不适用于过敏志愿者。 目的：确定过敏个体吸入低剂量内毒素（10,000 内毒素单位）（脂多糖 [LPS]）攻击的安全性，测量诱导痰 (IS) 后与抗原呈递和先天免疫相关的气道细胞表面表型。 LPS 攻击，并在 IS 细胞中进行基因表达谱分析，以确定哪些宿主遗传网络因 LPS 吸入而改变。方法：在 10 名过敏志愿者（8 名哮喘患者和 2 名鼻炎患者）吸入 LPS 之前和之后 6 小时获得诱导痰。使用流式细胞术检查 IS 细胞的细胞表面表型。使用微阵列和独创性途径分析对 IS 样本子集 (n = 10) 进行基因组表达分析。结果：总共 10,000 个内毒素单位的 LPS 诱导痰吞噬细胞上的膜 CD14、CD11b、CD16、HLA-DR、CD86 和 Fc epsilon 受体 1 显着上调，并增加影响抗原呈递表面分子（HLA-DR、趋化因子配体 2 或单核细胞趋化蛋白 1、v-rel 网状内皮细胞增生症病毒癌基因同源物、前列腺素内过氧化物合酶 2 或环氧合酶 2 和转化生长因子 β）、免疫激活（CD14、白细胞介素 1 β，以及激活后调节、正常 T 细胞表达和分泌）和炎症（细胞内粘附分子 1 和抑制性 kappa B）阿尔法）。核因子 kappa B、白细胞介素 1 和肿瘤坏死因子途径的基因谱也受到显着影响。结论：低剂量吸入内毒素攻击对于患有轻度至中度疾病的过敏个体是安全的。它增强气道细胞表面表型以及与抗原呈递、先天免疫和炎症相关的基因表达。具有独创性途径分析的微阵列可以成功地应用于痰细胞，以表征对吸入加剧剂的遗传反应。
Background: Environmental exposure to endotoxin is a known cause of exacerbation of asthma. Inhaled endotoxin protocols have been used to evaluate airway cell surface phenotypes associated with antigen presentation and innate immunity in healthy volunteers, but not in allergic volunteers.Objectives: To establish the safety of challenge with low-dose endotoxin (10,000 endotoxin units) (lipopolysaccharide [LPS]) inhalation in allergic individuals, to measure airway cell surface phenotypes associated with antigen presentation and innate immunity in induced sputum (IS) after LPS challenge, and to conduct gene expression profiling in IS cells to determine which host genetic networks are modified by LPS inhalation.Methods: Induced sputum was obtained before and 6 hours after LPS inhalation in 10 allergic volunteers (8 with asthma and 2 with rhinitis). Flow cytometry was used to examine cell surface phenotypes on IS cells. Genomic expression was analyzed on a subset of IS samples (n = 10) using microarray and ingenuity pathway analysis.Results: A total of 10,000 endotoxin units of LPS induced significant up-regulation of membrane CD14, CD11b, CD16, HLA-DR, CD86, and Fc epsilon receptor 1 on sputum phagocytes and increased expression of genes that influence antigen-presenting surface molecules (HLA-DR, chemokine ligand 2 or monocyte chemoattractant protein 1, v-rel reticuloendotheliosis viral oncogene homolog, prostaglandin-endoperoxide synthase 2 or cyclooxygenase 2, and transforming growth factor beta), immune activation (CD14, interleukin 1 beta, and regulated upon activation, normal T cell expressed and secreted), and inflammation (intracellular adhesion molecule 1 and inhibitory kappa B alpha). Gene profiles for nuclear factor kappa B, interleukin 1, and tumor necrosis factor pathways were also significantly affected.Conclusions: Low-dose inhaled endotoxin challenge is safe in allergic individuals with mild to moderate disease. It enhances airway cell surface phenotypes and expression of genes associated with antigen presentation, innate immunity, and inflammation. Microarray with ingenuity pathway analysis can be successfully applied to sputum cells to characterize genetic responses to inhaled exacerbants.