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

烟曲霉已成为人类最重要的空气传播真菌病原体，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。在感染过程中，病原真菌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。为了全面了解不同感染过程中存在的蛋白质以及对毒力可能重要的蛋白质，我们开始对烟熏假单胞菌进行蛋白质组分析。在这个优先项目中，我们的目标是生成烟曲霉在不同条件下生长的蛋白质组数据，包括真菌与巨噬细胞、中性粒细胞的共培养以及从感染小鼠中重新分离真菌。在此基础上，结合其他组的数据，建立蛋白质组数据库。通过蛋白质组分析确定的有趣基因将进行遗传和生化分析。将在巨噬细胞试验和低剂量吸入性小鼠感染模型中测试各自基因的缺失突变，以确定这些基因对毒力的贡献。将产生一种免疫原性转基因烟曲霉菌株，分析其在活鼠体内特异性免疫反应的发展。这些研究将导致鉴定对毒力重要的调控回路，更好地了解病原体和宿主之间的交流，并最终鉴定抗真菌药物的靶点。

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.