COCKROACH ALLERGENS--IMMUNOCHEMISTRY & ALLERGIC DISEASE

蟑螂(CR)侵袭房屋会产生蛋白质变应原，而蛋白质过敏原是一种 IgE介导的变态反应和哮喘的重要原因 少数族裔人口居住在不合标准的住房中。这样做的目的是 GRANT将克隆德国小蠊和大熊猫的过敏原 产生重组德国白僵菌变应原(Bla g 2、Bla g 4和Bla g 5)；建立免疫分析方法以评估接触CR； 制造不再结合IgE的过敏原分子；并分析T细胞 对CR过敏原的反应。这些目的将通过筛选cDNA来实现 含IgE抗体(Ab)的文库和产生重组变应原的 表达载体。重组Bla-g2、Bla-g4的生物学活性 &Bla g 5将通过皮肤测试、组胺释放和血清进行比较 在一项关于哮喘患者的多中心研究中进行免疫球蛋白单抗检测 夏洛茨维尔、新奥尔良、巴尔的摩、奥古斯塔、佐治亚州和斯特拉斯堡， 法国。房屋中接触CR过敏原的情况将使用以下方法进行评估 基于单抗的CR感染的检测和计数。这些 实验将集中在开发P。 美洲变应原，以及德国伯氏杆菌，以确定变应原 粉尘样本中导致IgE过敏和症状的水平。另外， 目的：研究CR变应原的空气动力学特性。站点定向 突变将被用来定位Bla g 4和Bla g 5上的IgE ab位点 并产生减少与IgE结合的结构变化。氨基 将通过分子模拟将酸作为诱变的目标。T细胞 对重组变应原的反应将通过增殖来比较 检测以及对每种过敏原的反应的流行率和大小将 要下定决心。将使用变应原变体绘制T细胞位置图， 合成肽或表达的部分变应原序列。这笔赠款 将加深我们对特定CR的作用和重要性的理解 引起哮喘的过敏原。变态反应原的产生 突变，结合T细胞反应的分析，也可能提供 CR哮喘的免疫治疗策略更加合理。

Cockroaches (CR) infesting houses produce protein allergens which are an important cause of IgE mediated allergic reactions and asthma among minority populations living in sub-standard housing. The aims of this grant are to clone allergens from Blattella germanica and Periplaneta americana; to produce recombinant B. germanica allergens (Bla g 2, Bla g 4 and Bla g 5); to develop immunoassays to assess exposure to CR; to produce allergen molecules that no longer bind IgE; and to analyze T cell responses to CR allergens. These aims will be achieved by screening cDNA libraries with IgE antibodies (ab) and producing recombinant allergens in expression vectors. The biologic activity of recombinant Bla g 2, Bla g 4 & Bla g 5 will be compared by skin testing, histamine release and serum IgE ab assays in a multi-center study of patients with asthma enrolled in Charlottesville, New Orleans, Baltimore, Augusta, GA, and in Strasbourg, France. Exposure to CR allergens in houses will be assessed using monoclonal antibody based assays and counts of CR infestation. These experiments will focus on developing tests for P. americana allergens, as well as B. germanica, in order to define allergens levels in dust samples which cause IgE sensitization and symptoms. Also, to investigate the aerodynamic properties of CR allergens. Site directed mutagenesis will be used to localize IgE ab sites on Bla g 4 and Bla g 5 and to generate structural alterations that reduce binding to IgE. Amino acids will be targeted for mutagenesis by molecular modeling. T cell responses to recombinant allergens will be compared by proliferation assays and the prevalence and magnitude of responses to each allergen will be determined. T cell sites will be mapped using allergen variants, synthetic peptides or expressed partial allergen sequences. This grant will increase our understanding of the role and importance of specific CR allergens in causing asthma. The production of altered allergens by mutagenesis, combined with analyses of T cell responses, may also provide more rational strategies for immunotherapy of CR asthma.