COCKROACH ALLERGENS--IMMUNOCHEMISTRY & ALLERGIC DISEASE

越来越多的证据表明，蟑螂（CR）致敏是一种 哮喘的重要原因，特别是在非洲裔美国人中， 在城市地区的CR出没的住房。 当前的目标 我们的建议是识别，纯化和序列的主要过敏原， 两种最常见的室内CR物种，德国小蠊和大蠊 美国。 鼠单克隆抗体（mAb）将针对 CR提取物并筛选以鉴定人IgE识别的抗原 抗体的 选定的mAb将与生物化学试剂盒一起使用。 纯化CR过敏原的技术。 重组DNA技术将 用于确定主要CR的完整核苷酸序列 过敏原 序列将从聚合酶获得， 链反应（PCR）扩增产物或来自CR cDNA克隆， 将使用寡核苷酸探针、mAb或IgE进行鉴定 抗体的 将PCR产物或cDNA克隆进行亚克隆， 通过双脱氧测序进行测序。 过敏原编码cDNA将被 克隆到细菌载体中，以促进 重组过敏原。 这些研究将确定结构 以及不同CR产生的变应原之间的抗原关系 物种将使用序列相似性搜索来研究 CR变应原的生物学功能及免疫细胞化学技术 将用于定位CR组织中过敏原产生的位点。 基于单克隆抗体的免疫测定将用于测量室内CR过敏原 哮喘患者家中灰尘和空气样本的含量 以及非哮喘对照组的那些。初步研究表明， 在弗吉尼亚州夏洛茨维尔、佐治亚州亚特兰大和特拉华州威尔明顿进行。 的 拟议的研究将调查过敏原积聚的部位， 空气中的颗粒大小和浓度 过敏原目的是建立CR过敏原的形式， 引起IgE抗体反应致敏并确定阈值 暴露水平是致敏和 过敏性疾病的恶化。 这些试验将用于 调查控制CR感染的方法是否会导致 降低CR过敏原水平。 这些研究将有助于建立 在哮喘管理中使用CR控制的可行性。 的 拟议的研究旨在提高对一组 过敏原似乎是哮喘的重要原因，特别是 在那些受最近 这种疾病的死亡率和发病率增加。

Increasing evidence suggests that cockroach (CR) sensitization is an important cause of asthma, particularly among African-Americans living in urban areas in CR infested housing. The aims of the present proposal are to identify, purify and sequence major allergens from the two most common indoor CR species, Blattella germanica and Periplaneta americana. Murine monoclonal antibodies (mAb) will be raised against CR extracts and screened to identify antigens recognized by human IgE antibodies. Selected mAb will be used together with biochemical techniques to purify CR allergen(s). Recombinant DNA techniques will be used to determine the complete nucleotide sequences of major CR allergens. The sequences will be obtained either from polymerase chain reaction (PCR) amplification products or from CR cDNA clones, which will be identified using oligonucleotide probes, mAb or IgE antibodies. PCR products or cDNA clones will be sub-cloned and sequenced by dideoxy sequencing. Allergen encoding cDNA will be cloned into bacterial vectors to facilitate high level expression of recombinant allergens. These studies will determine the structural and antigenic relationships between allergens produced by different CR species. Sequence similarity searches will be used to investigate the biologic function of CR allergens and immunocytochemical techniques will be used to localize sites of allergen production in CR tissues. MAb based immunoassays will be used to measure indoor CR allergen levels in dust and air samples from the houses of patients with asthma and in those of non-asthmatic controls. Preliminary studies have been carried out in Charlottesville VA, Atlanta GA and Wilmington DE. The proposed studies will investigate the sites of allergen accumulation in the home and the particle size and concentration of airborne allergen. The aim will be to establish the form in which CR allergens cause sensitization for IgE antibody responses and to define threshold levels of exposure that are risk factors for both sensitization and exacerbations of allergic disease. The assays will be used to investigate whether methods to control CR infestation can cause reductions in CR allergen levels. These studies would help establish the feasibility of using CR control in the management of asthma. The proposed studies are designed to improve understanding of a group of allergens that appear to be an important cause of asthma, particularly among those minority populations that are most affected by the recent increases in mortality and morbidity from this disease.