Identification of virulence determinants of the human-pathogenic fungus Aspergillus fumigatus by proteome analysis

烟曲霉菌已成为人类最重要的气载真菌病原体，在免疫功能低下的宿主中引起肺炎和侵袭性播散性疾病，死亡率高。病原真菌在感染过程中需要应对各种环境挑战。应对这些挑战的能力需要许多基因的表达。为了全面了解在不同感染过程中存在的蛋白质，并对毒力有潜在的重要性，我们已经开始建立A.烟熏。在这个优先项目中，我们的目标是产生A.在不同条件下生长的烟曲霉，包括真菌与巨噬细胞、嗜中性粒细胞的共培养以及从感染的小鼠中重新分离真菌。基于这些数据以及来自其他组的数据，将建立蛋白质组数据库。通过蛋白质组分析确定的感兴趣的基因将进行遗传和生物化学分析。将在巨噬细胞试验和低剂量吸入性小鼠感染模型中检测相应基因的缺失突变体，以确定这些基因对毒力的贡献。免疫原性转基因A.将产生烟曲霉菌株以分析活小鼠中特异性免疫反应的发展。这些调查将导致重要的毒力，更好地了解病原体和宿主之间的通信，并最终确定抗真菌药物的目标的监管电路的识别。

Aspergillus fumigatus has become the most important airborne fungal pathogen of humans causing pneumonia and invasive disseminated disease with high mortality in the immunocompromised host. Pathogenic fungi need to deal with a variety of environmental challenges during the course of an infection. The ability to meet these challenges requires the expression of many genes. To obtain a comprehensive overview about the proteins present during different infectious processes and potentially important for virulence, we have started to establish proteome analysis for A. fumigatus. Within this priority programme we aim at generating proteome data of A. fumigatus growing under different conditions including the cocultivation of the fungus with macrophages, neutrophils and the reisolation of the fungus from infected mice. Based on these data together with data from other groups, a proteome data base will be established. Interesting genes identified by proteome analysis will be analysed both genetically and biochemically. Deletion mutants of the respective genes will be tested in both macrophage assays and in a low dose, inhalative mouse infection model to determine the contribution of these genes to virulence. An immunogenic transgenic A. fumigatus strain will be generated to analyse the development of a specific immune reaction in living mice. These investigations will lead to the identification of regulatory circuits important for virulence, a better understanding of the communication between pathogen and host and eventually to the identification of targets for antifungal drugs.