MODULATION OF THE HUMAN IGE RESPONSE BY XENOBIOTICS

该项目将确定芳烃， 例如空气污染中那些，增强人过敏性抗体（IgE）， 反应，从而有助于显着增加过敏性 这是世纪发生的呼吸道疾病 流行病学数据 这表明环境因素在其中起着重要作用。 增加过敏性疾病的表达。 然而，几乎没有人注意到 直接测试环境影响，例如 柴油机废气中的多环芳烃（PAH- DEP），特异性IgE反应后也不以基本机制通过 这可能发生。 关于IgE调节的知识和注意事项 现有的流行病学数据引导我们进行开创性的实验 发现PAH-DEP对人IgE具有直接的免疫增强作用 PAH-DEP可以协同从B细胞产生IgE， 以及改变T和B细胞表面分子的表达， IgE产生。 本项目将研究PAH-DEP如何调节定义的 IgE产生途径中的步骤，从而确定机制 PAH-DEP通过其定量地改变IgE反应（总的量 IgE蛋白）和定性（IgE蛋白6种亚型的相对量）。 通过替代产生的β链[4条分泌链和2条膜链] 拼接[我们描述]）。 具体来说，我们将#1确定 PAH-DEP对NB细胞的直接作用a）表面分子/受体 与IgE合成相关，B）IgE mRNA转录水平和 剪接变体和c）膜和分泌的IgE同种型的量 制作。 这涉及刺激人类B细胞通过定义的阶段 在IgE产生中（IgE种系转录，IgE同种型 转换和调节生产性PAH mRNA）和定义PAH- DEP改变了这些过程和结果。 我们将#2）评估PAH-DEP， 改变T细胞提供的激活/接触信号的能力 (and T细胞亚群）参与IgE的产生。 这既包括 流式细胞仪表型研究和功能研究 PAH-DEP处理T细胞在IgE产生中协作的能力。 第三名） PAH-DEP改变参与的关键细胞因子产生的能力 在IgE产生途径中（例如IL-4、TGF β、IL-6等）将 在蛋白质和mRNA水平上进行检测。 我们将把这些 通过#4a）评估柴油机排气颗粒（DEP）影响的体内研究 对低芳烃和高芳烃中体内IgE抗体应答的影响 水解酶活性（Ah活性）小鼠。 这些研究将证明， PAH-DEP通过常见的Ah受体复合物起作用， 为模型，以扩大研究的动力学暴露于多环芳烃-DEP。 我们 还将进行#4b）局部给药的体内人体挑战研究 DEP对IgE定量和定性表达的影响 上呼吸道的反应。 我们的研究将提供洞察力， 可能是影响变态反应性改变的关键因素之一， 这是世纪发生的呼吸道疾病

This project will determine the mechanisms which aromatic hydrocarbons, such as those in air pollution, enhance human allergic antibody (IgE) responses and thereby contribute to the marked increase in allergic airway disease that has occurred this century. Epidemiologic data suggest that environmental factors play an important role in the increasing expression of allergic disease. Yet almost no attention has been given to directly testing if environmental influences, exemplified by the polycyclic aromatic hydrocarbons found in diesel exhaust (PAH- DEP), specifically after the IgE response nor to the basic mechanism by which this may occur. Knowledge about IgE regulation and consideration of the available epidemiologic data lead us to the seminal experimental discovery that PAH-DEP has direct immuno-enhancing effects on human IgE production; PAH-DEP can synergize in production of IgE from B cells as well as alter expression of T and B cell surface molecules relevant to IgE production. This project will examine how PAH-DEP modulates defined steps in pathways for IgE production and thereby determine the mechanisms by which PAH-DEP alter the IgE response quantitatively (total amount of IgE protein) and qualitatively (relative amount of the 6 isoforms of epsilon chains [4 secreted and 2 membrane] generated by alternative splicing [described by us]). Specifically we will #1 determine the direct effects of PAH-DEP on NB cell a) surface molecules/receptors relevant to IgE synthesis, b) epsilon mRNAs transcript levels and splicing variants and c) the amount of membrane and secreted IgE isoforms produced. This involves stimulating human B cells through defined stages in IgE production (epsilon germ line transcription, epsilon isotype switch and modulation of productive epsilon mRNA) and defining how PAH- DEP alters these processes and outcomes. We will #2) assess PAH-DEP for the ability to modify the activation/contact signals provided by T cells (and T cell subsets) involved in IgE production. This includes both multicolor flow cytometric phenotypic studies and functional studies of PAH-DEP treated T cells' ability to collaborate in IgE production. #3) The ability to PAH-DEP to alter the production of key cytokines involved in pathways for IgE production (e.g. IL-4, TGFbeta, IL-6, etc.) will be examined at both the protein and MRNA levels. We will extend these studies in vivo by #4a) assessing diesel exhaust particle (DEP) effects on the in vivo IgE antibody response in low and high aromatic hydrocarbon hydrolase activity (Ah activity) mice. These studies will prove whether PAH-DEP is working via the common Ah receptor complex as well as provide for a model to extend studies of kinetics of exposure to PAH-DEP. We will also undertake #4b) in vivo human challenge studies of the topical effects of DEP on the expression of the quantitative and qualitative IgE response in the upper airway. Our studies will provide insight into what may be one of the critical factors affecting the change in allergic airway disease that has occurred this century.