MOLECULAR AND CELL BIOLOGY OF FUNGAL PATHOGENS

在这个项目中，我们将进行医学方面的基础研究 重要的真菌在分子和细胞水平上：1)增强 对真菌致病机理的认识；2)建立真菌的生化标志物 真菌感染；3)探索人类吞噬细胞表面受体 与组织胞浆菌(HC)配体的相互作用及其后续作用 吞噬细胞内生化事件的信号；4)进一步表征 卡氏肺孢子虫的基因结构和组织 最近通过小亚基证明是真菌的一员 核糖体RNA比较分析。项目。%1将制作和描述 核酸探针分析基因(S)和其他同源蛋白 负责编码HC酵母中的质膜质子泵 保持体内pH值合理恒定，并可能提高存活率 在宿主巨噬细胞吞噬酶体的酸性环境中。项目。2个将会 检测组织胞浆酵母菌和微孢子在人体内的命运 单核/巨噬细胞和肺泡巨噬细胞。酵母菌中的配体 和分生孢子，由LFA-1，CR3，p150，95受体识别 巨噬细胞将被鉴定和纯化。信号机制 我们将比较酵母与这些受体结合后启动的转导。 而不是C3bi包被红细胞与CR3结合所致。项目。3. 将研究弹性蛋白水解酶在猪瘟病毒毒力中的作用 分离和克隆侵袭性曲霉菌的基因 曲霉弹性蛋白酶(AE)。克隆的AE基因将用于生产 用于诊断试验和毒力研究的重组AE； 将确定基因的染色体位置，并将AE基因探针 将通过原位杂交进行诊断评估。项目。4个人会 追踪观察曲霉和隐球菌的致病菌种 产生大量的多元醇，D-甘露醇。毛细管 将使用气相色谱来确定这种代谢物是否可以 作为侵袭性感染的定量诊断标志 这些真菌存在于实验动物和患者体内。项目。5将延长我们的 利用克隆DNA对卡氏肺孢子虫真菌性质的认识 用于表征基因结构和组织的片段。编码基因 核糖体RNA和钙调蛋白基因将是主要的克隆目标。 RRNA基因、染色体标记和重复元件的克隆将是 用作杂交探针检测猪瘟病毒分离株的基因组变异 鼠类和人类宿主的卡氏肺孢子虫。

In this program project we will conduct basic studies on medically important fungi at the molecular and cellular level to: 1) enhance understanding of fungal pathogenesis; 2) develop biochemical markers of infection by fungi; 3) explore human phagocyte surface receptor interactions with ligands of Histoplasma capsulatum (Hc) and the subsequent signaling of intraphagocyte biochemical events; 4) further characterize the gene structure and organization of Pneumocystis carinii which we and others recently demonstrated to be a member of the fungi by small subunit ribosomal RNA comparative analysis. Proj. 1 will produce and characterize nucleic acid probes to analyze the gene(s) and other cognate proteins responsible for encoding a plasma membrane proton ion pump in Hc yeasts that maintains a reasonably constant internal pH and may enhance survival within the acidic milieu of host macrophage phagolysosomes. Proj. 2 will examine the fate of Histoplasma yeasts and microconidia in human monocyte/macrophages and alveolar macrophages. The ligands within yeasts and conidia that are recognized by the LFA-1,CR3,p150,95 receptors of macrophages will be identified and purified. Mechanisms of signal transduction initiated by yeast binding to these receptors will be compared to those induced by binding of C3bi-coated erythrocytes to CR3. Proj. 3 will study the role of elastinolytic proteinase in the virulence of invasive Aspergillus species by isolating and cloning the gene for Aspergillus elastase (AE). Cloned AE gene will be used to produce recombinant AE for use in diagnostic testing and virulence studies; the chromosomal location of the gene will be determined, and AE gene probes will be evaluated for diagnosis by in situ hybridization. Proj. 4 will pursue observations that pathogenic species of Aspergillus and Cryptococcus produce large amounts of the polyol, D-mannitol. Capillary gas-chromatography will be used to determine if this metabolite can be employed as a quantitative, diagnostic marker of invasive infection by these fungi in experimental animals and patients. Proj. 5 will extend our knowledge of the fungal nature of P. carinii by employing cloned DNA fragments to characterize gene structure and organization. Genes encoding ribosomal RNAs and the calmodulin gene will be primary cloning targets. Clones of rRNA genes, chromosome markers and repetitive elements will be used as hybridization probes to examine genomic variation among isolates of P. carinii from rat and human hosts.