ANTIGENIC STRUCTURE OF PURIFIED HOUSE DUST MITE ALLERGEN

屋尘螨（Dermatophagoides）是引起 过敏性疾病，如常年性鼻炎、哮喘和特应性 dermatitis和两个主要种D. pteronyssinus和D. 在美国的许多地方， States. 该项目的目的是确定主要的 结构和两种主要抗原之间的抗原关系 尘螨过敏原的组：24kD组I过敏原（Derp I， Der f I和Der m I）和15kd II类过敏原（Der p II和Der m I）。 Der f II）。 这些过敏原的氨基酸序列将是 使用肽测序的组合方法确定 （Edman降解和串联质谱法）和cDNA 在λ gt 10中克隆。 结构同源性 已知氨基酸序列过敏原和其它蛋白质， 比较了 鼠单克隆抗体和人单克隆抗体的结合 将比较两个过敏原组的IgG和IgE抗体 通过抗原结合放射免疫测定。 表位作图研究 将通过比较抗体与变性的 过敏原、化学修饰的过敏原和肽片段。 目的是确定B细胞表位的所有组成部分， 它们是连续的还是局部的， 残基提供抗体结合的接触点。 鼠 对I组过敏原的抗体应答将在 使用不同免疫方案的几种近交系， 研究是否影响同种型的免疫模式 表达也影响表位特异性。 人T细胞 将比较对I组和II组过敏原的反应， 利用来自螨的外周血T细胞的体外细胞培养系统 过敏患者 合成肽将用于刺激T 细胞克隆和鉴定T细胞表位。 抗原的作用 呈递细胞在呈递给T细胞之前处理过敏原 细胞也将被研究。 这项建议提供了一个系统的方法来研究 主要尘螨过敏原的分子和抗原结构。 这些研究应能使人们更好地了解 其影响体液和细胞免疫应答， 螨过敏原。 它们还应提供进一步的知识， 为什么这些过敏原是鼻炎，哮喘的常见原因 和特应性皮炎。

House dust mites (Dermatophagoides) are an important cause of allergic diseases such as perennial rhinitis, asthma and atopic dermatitis and the two principal species, D. pteronyssinus and D. farinae have been found in homes in many parts of the United States. The aims of this project are to determine the primary structures and the antigenic relationships between the two major groups of dust mite allergens: the 24kd Group I allergens (der p I, Der f I, and Der m I) and the 15kd Group II allergens (Der p II and Der f II). The amino acid sequences of these allergens will be determined using the combined approaches of peptide sequencing (Edman degradation and tandem mass spectrometry) and cDNA cloning in Lambda gt 10. Structural homology between each allergen and other proteins of known amino acid sequence will be compared. Binding of murine monoclonal antibodies and human IgG and IgE antibodies to both allergen groups will be compared by antigen binding radioimmunoassays. Epitope mapping studies will be carried out by comparing antibody binding to denatured allergens, chemically modified allergens and peptide fragments. The aim will be to determine the repertoire of B cell epitopes, whether they are sequential or topographic and which amino acid residues provide the contact points for antibody binding. Murine antibody responses to Group I allergens will be compared in several inbred strains using different immunization regimes to investigate whether modes of immunization which affect isotype expression also influence epitope specificity. Human T cell responses to the Group I and II allergens will be compared n in vitro cell culture systems using peripheral blood T cells from mite allergic patients. Synthetic peptides will be used to stimulate T cell clones and to identify T cell epitopes. The role of antigen presenting cells in processing allergens prior to presentation to T cells will also be investigated. This proposal provides a systematic approach to studying the molecular and antigenic structure of major dust mite allergens. These studies should result in greater understanding of factors which influence both humoral and cellular immune responses to mite allergens. They should also provide further knowledge as to why these allergens are such a common cause of rhinitis, asthma and atopic dermatitis.