COCKROACH ALLERGENS--IMMUNOCHEMISTRY & ALLERGIC DISEASE

蟑螂（CR）侵扰房屋产生蛋白质过敏原， IgE介导的过敏反应和哮喘的重要原因， 少数民族居住在低于标准的住房中。其目的是 格兰特将从德国小蠊和大蠊中克隆过敏原 americana;以产生重组B。德国变应原（Bla g 2、Bla g 4和Bla g 5）;开发免疫测定法以评估CR暴露; 产生不再结合IgE的过敏原分子;并分析T细胞 对CR过敏原的反应。这些目标将通过筛选cDNA来实现 文库与IgE抗体（ab）和生产重组过敏原， 表达载体。重组Bla g 2、Bla g 4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 一项多中心哮喘患者研究中的IgE抗体检测 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 将使用以下方法评估房屋中CR过敏原的暴露 基于单克隆抗体的测定和CR侵染计数。这些 实验将集中于开发P的测试。 美国过敏原，以及B。germanica，为了定义过敏原 粉尘样本中的IgE水平会导致过敏和症状。还有， 研究CR过敏原的空气动力学特性。定点 将使用诱变来定位Bla g 4和Bla g 5上的IgE ab位点 并产生减少与IgE结合的结构改变。氨基 通过分子模拟将酸作为诱变的靶。t细胞 对重组变应原的反应将通过增殖 对每种过敏原的反应的流行率和程度将 下定决心。T细胞位点将使用过敏原变体进行定位， 合成肽或表达的部分变应原序列。 这笔赠款 将增加我们对特定CR的作用和重要性的理解 过敏原引起哮喘。过敏原的改变， 诱变，结合T细胞反应的分析，也可以提供 CR哮喘免疫治疗的合理策略。

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.