Cytokine profiles of BAL T cells and T-cell clones obtained from human asthmatic airways after local allergen challenge
Cytokine profiles of BAL T cells and T-cell clones obtained from human asthmatic airways after local allergen challenge
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DOI：
10.1034/j.1398-9995.1999.00889.x
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发表时间：
1999-10-01
期刊：
影响因子：
12.4
通讯作者：
Frew, AJ
中科院分区：
文献类型：
--
作者：
Bodey, KJ;Semper, AE;Frew, AJ
背景：本研究评估了局部过敏原激发前后哮喘细胞因子表达的异质性。方法：BAI。在特应性哮喘受试者局部支气管内变应原激发后10分钟或24小时获得T细胞。通过直接有限稀释法克隆T细胞。mRNA表达通过RT-PCR评估，细胞因子蛋白生产通过ELISA.Results：未刺激的基线BAL T细胞表达IFN-λ、IL-13和TNF-α的mRNA。少数样本表达IL-4和IL-5，但未检测到IL-3 mRNA。PHA刺激在4/6个样品中增加IL-3、IL-4和IL-5 mRNA的表达。IL-13和CM-CSF mRNA在BAL中表达。细胞在过敏原攻击后，但IFN-γ的表达减少。与等效基线样品相比，PHA刺激后IL-4和IL-3均强烈上调，而TNF-α和IFN-γ的表达降低。得到17组BAL I细胞克隆（平均克隆效率1/40 T细胞）。7个板存活至8周用于分析。盐水攻击后4小时衍生的克隆显示IL-13、IL-4和IFN-γ的强mRNA信号，而过敏原攻击后24小时衍生的克隆表达IL-13、CM-CSF、IL-3、IL-4和通常IL-5（即，更接近Th 2分布）。有相当大的异质性的细胞因子的mRNA和蛋白质的生产模式由不同的clones.Conclusions：T细胞从哮喘气道产生IL-13，IFN-γ，和TNF-α，但过敏原的挑战后，2型细胞因子上调。mRNA和蛋白质分析为气道T细胞细胞因子谱提供了补充信息。
Background:This study assessed the heterogeneity of cytokine expression in asthma before and after local allergen challenge. Methods: BAI. T cells were obtained 10 min or 24 h after local endobronchial allergen challenge in atopic asthmatic subjects. T cells were cloned by direct limiting dilution. mRNA expression was assessed by RT-PCR, and cytokine protein production by ELISA.Results: Unstimulated baseline BAL T cells expressed mRNA for IFN-lambda, IL-13, and TNF-alpha. A minority of samples expressed IL-4 and IL-5, but no IL-3 mRNA was detected. PHA stimulation increased expression of IL-3, IL-4 and lL-5 mRNA in 4/6 samples. IL-13 and CM-CSF mRNA were found in BAL. cells after allergen challenge, but expression of IFN-gamma was reduced. Both IL-4 and IL-3 were strongly upregulated after PHA stimulation, while the expression of TNF-alpha and IFN-gamma was reduced, compared to equivalent baseline samples. Seventeen panels of BAL I-cell clones were derived (average cloning efficiency 1/40 T cells). Seven panels survived to 8 weeks for analysis. Clones derived 4 h after saline challenge showed strong mRNA signals for IL-13, IL-4, and IFN-gamma, whereas clones derived 24 h after allergen challenge expressed IL-13, CM-CSF, IL-3, IL-4 and often IL-5 (i.e., closer to the Th2 profile). There was considerable heterogeneity in the patterns of cytokine mRNA and protein production by different clones.Conclusions: T cells from asthmatic airways produce IL-13, IFN-gamma, and TNF-alpha, but after allergen challenge, type 2 cytokines are upregulated. mRNA and protein analysis provide complementary information on airways T-cell cytokine profiles.