ASPERGILLUS ALLERGENS AND ALLERGIC ASPERGILLOSIS

描述(改编自申请人的摘要)：总体目标 该项目的主要目的是生产和鉴定相关的曲霉 烟曲霉菌(Af)抗原对曲霉菌的早期诊断和了解 过敏性曲霉病的免疫发病机制。过敏性支气管肺 曲霉病(Aspergillosis，ABPA)是一种破坏性的肺部疾病，常见于 特应性个体和囊性纤维化患者。本病的诊断 疾病主要是基于免疫学标准，但我们对 该病的免疫诊断和免疫发病机制为 不能令人满意。此外，由于缺乏可靠的试剂和 检测的重复性不一致，对检测的解释 结果经常是困难的，贡献较小，并已被引用 作为更多患者在早期发病期间未被诊断的主要原因 这种疾病的危害。用分子生物学方法从Af中获得四种纯抗原 克隆已经确定，所有的克隆都显示出显著的免疫球蛋白E ABPA的抗体结合和T细胞增殖活性。两者都在体外 体内试验将用于证明Af特异性IgE抗体在 使用四种重组Af抗原的患者的血清。表位 将确定与IgE的结合并与其他已知过敏原进行比较 独一无二的基因将被克隆和表达。这些克隆的表位含有 多肽的特异性和相关性将在 ABPA的诊断。ABPA患者的T细胞将被克隆用于 通过研究Th1和Th2来了解该病的发病机制 细胞因子图谱。针对Asp f2产生的T细胞系将被刺激 与来自重组蛋白Asp f2序列的合成肽 与ABPA和T细胞表位的反应性将被确定。这个 这些克隆产生的细胞因子对合成的T细胞表位的反应 将进行评估，以了解疾病的发病机制。这个 细胞因子图谱将增加我们对病理生理事件的了解 与疾病相关的。这些研究的结果将提供更多的 需要了解ABPA中活跃的免疫机制，在未来 可以用来有效地控制或管理疾病。

DESCRIPTION (Adapted from the applicant's abstract): The overall objectives of this project are to produce and characterize relevant Aspergillus fumigatus (Af) antigens for early diagnosis of Aspergillus and to understand the immunopathogenesis of allergic aspergillosis. Allergic bronchopulmonary aspergillosis (ABPA) is a destructive lung disease frequently occurring in atopic individuals and in cystic fibrosis patients. The diagnosis of the disease is based mostly on immunological criteria, yet our understanding of the immunodiagnosis and immunopathogenesis of this disease is unsatisfactory. Furthermore, due to the lack of reliable reagents and inconsistent reproducibility of assays, the interpretation of the test results are frequently difficulty and less contributory and has been cited as a major reason why more patients are not diagnosed during the early onset of the disease. Four pure antigens from Af obtained through molecular cloning have been identified and all of them demonstrated significant IgE antibody binding and T-cell proliferation activities in ABPA. Both in vitro and in vivo tests will be used to demonstrate Af specific IgE antibodies in the sera of patients using the four recombinant Af antigens. The epitopes binding to IgE will be identified and compared with other known allergens and unique ones will be cloned and expressed. These cloned epitopes bearing polypeptides will be evaluated for their specificity and relevancy in the diagnosis of ABPA. T-cells from ABPA patients will be cloned for understanding the mechanism of the disease by studying the Th1 and Th2 cytokine profile. T-cell lines generated against Asp f2 will be stimulated with synthetic peptides from the sequence of Asp f2, a recombinant protein with reactivity with ABPA< and T-cell epitopes will be identified. The cytokines produced by these clones in response to synthetic T-cell epitopes will be assessed to understand the pathogenesis of the disease. The cytokine profile will add to our knowledge on the pathophysiological events associated with disease. The results of these studies will provide the much needed understanding of the immune mechanism active in ABPA, which in future can be used to control or manage the disease effectively.