Impact of allergy and seasonality on lymphokine producing T cells

项目总结(见说明)： RFA的两个主要目标是对来自大规模变应原表位识别合同的表位进行表征，1)作为季节性的函数，2)疾病进展。因此，我们建议检验这一假设，即不同的阶段(季节性与非季节性)、类型(鼻炎与哮喘)和过敏性疾病的严重程度不仅与T细胞反应的不同程度有关，而且还与所涉及的TH亚群及其相互作用的不同模式相关。我们将重点关注三种重要过敏原的免疫优势区域，即蒂莫西草(Tg)、屋尘螨(HDM)和猫皮屑(CD)。在我们的第一个目标中，我们建议将反应描述为中等/严重 (Mo/SA)和轻度哮喘患者(MA)，并与非哮喘患者进行对照 患有过敏性鼻炎(AR)。初步数据显示，ILIO、ILI7和IFNY产量不同 与不同的疾病状态相关。在这里，我们建议刻画产生TH细胞的子集 这些淋巴因子在体外和体外刺激后，通过ELISPOT细胞内细胞因子染色(ICS) 四聚体染色分析。我们将进一步检查不同的子集是不同的还是可以 在体外彼此起源，因为TH细胞的可塑性。初步数据还表明， 当比较TG和HDM反应时，TH亚集的平衡有很大不同。因此，我们将 研究TH亚群的不同分布是否与变应原来源相关。我们的初步数据 进一步强调作为季节性暴露的函数，TG反应的大小存在显著差异。在第二个目标中，我们建议在纵向研究中确定反应幅度和TH功能子集。在进一步的一系列实验中，我们将利用甘油三酯花粉提取物进行挑战研究。我们预计，由于TH细胞的反应更强烈，在体外可能更容易检测到，而且 具有更详细的特点，不会因体外培养而引起潜在的变化。这些研究将表征不同疾病背景下的功能性T细胞亚群，并作为季节性的函数。

PROJECT SUMMARY (See instructions): Two of the RFA main goals are the characterization of epitopes derived from the Large Scale Allergen Epitope Identification Contracts, 1) as a function of seasonality and 2) disease progression. Accordingly, we propose to test the hypothesis that different stages (in-season versus out-of-season), types (rhinitis versus asthma) and severities of allergic disease are associated with not only differential magnitude of T cell responses, but also distinctive patterns in terms of the TH subsets involved and their interplay. We will focus on immunodominant regions from three important allergens, namely timothy grass (TG), house dust mite (HDM) and cat dander (CD). In our first Aim, we propose to characterize responses in moderate/severe (MO/SA) and mild asthmatics (MA), and contrast them with those observed in non-asthmatic individuals suffering from allergic rhinitis (AR). Preliminary data suggest that differential ILIO, ILI7 and IFNy production correlate with different disease states. Here, we propose to characterize the subsets of TH cells producing these lymphokines ex vivo and following in vitro stimulation, by ELISPOT, intracellular cytokine staining (ICS) and tetramer staining assays. We will further examine whether the different subsets are distinct or can originate from each other in vitro, because of TH cell plasticity. Preliminary data also suggest that the relative balance of TH subsets differs substantially when TG and HDM responses are compared. We will therefore investigate whether different distributions of TH subsets correlate with allergen source. Our preliminary data further highlights dramatic differences in magnitude of TG responses as a function of seasonal exposure. In the second Aim we propose to determine in longitudinal studies response magnitude and TH functional subsets. In a further series of experiments we will perform challenge studies utilizing TG pollen extracts. We expect that because of a more vigorous response TH cells might be more easily detectable ex vivo, and characterized in greater detail without potential alterations introduced by in vitro culture. These studies will characterize the functional T cell subsets involved in different disease settings and as a function of seasonality.