INFLUENZA VIRUS EFFECTS ON MACROPHAGE VESICLE TRAFFIC

流感病毒对巨噬细胞囊泡运输的影响

基本信息

批准号：
2071858
负责人：
ROBERT J LOWY
金额：
$ 6.81万
依托单位：
HENRY M. JACKSON FDN FOR THE ADV MIL/MED
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-05-01 至 1998-07-31
项目状态：
已结题

项目摘要

New and improved anti-microbial therapies are needed due to increasing immunocompromise populations and increasing spread of drug-resistant strains of microorganisms. Phagocytes (PC), important in pathogen clearance, can be inhibited so effectively by pathogens that virulence increases as PCs become the infectious agent reservoir and/or as susceptibility to secondary infections rises. Phagocytosis and endocytosis are among the important cellular functions suppressed by many notorious human pathogens, including influenza virus (IV), M. leprae and M. tuberculosis. These intracellular vesicle-based transport processes are central to both normal PC function and to most therapies, including drug delivery, modulation of activation and antigen processing. Not enough is known about either the specific phagosomal-endosomal processes affected or the physiological and biochemical mechanisms pathogens utilize to rationally design interventions to restore PC function. The major goal of this proposal is to determine what physiological changes occur in which phagosome-endosome processes resulting in failure of these anti-microbial cellular activities during IV interaction with a well-characterized phagocytic cell the J774.1 murine macrophage. Information on macrophage- pathogen effects is lagging that for other PCs, but during immunosuppression they can become the predominate remaining PC. IV is an important disease and many of its effects on PCs are similar to those of bacteria, but it is simpler and therefore the prospect of identifying important PC inhibitor mechanism(s) and improving anti-viral therapies is increased. The specific aims are: 1) Identify the extent and type of changes in macrophage intracellular trafficking pathways after exposure to IV. 2) Determine what changes occur in the intracellular physical-chemical environment of macrophages and intracellular compartments during early influenza virus exposure. 3) Investigate the effects of agents which either mimic or antagonize the IV-caused alterations in macrophage endosome and phagosome function. Quantitative digital video microscopy will be used to examine effects of IV on the J774 intracellular vesicle- based transport functions including the uptake and movement between compartments of molecular markers for specific endocytic and phagocytic functions. fluorescent probes will also be used to monitor the physical- chemical state of phagosomes and endosomes. It is expected that both the methodologies and cell biological findings will be applicable to understanding PC inhibition by other virus types, bacteria and parasites.

由于越来越多的细菌感染，需要新的和改进的抗微生物疗法免疫功能低下的人群和耐药性的日益蔓延微生物菌株。吞噬细胞（PC），在病原体中很重要清除率，可以被病原体如此有效地抑制，以致毒力随着 PC 成为传染源储存库和/或继发感染的易感性增加。吞噬作用和内吞作用是被许多臭名昭著的人抑制的重要细胞功能之一人类病原体，包括流感病毒 (IV)、麻风分枝杆菌和麻风分枝杆菌。结核。这些基于细胞内囊泡的运输过程是对于正常 PC 功能和大多数治疗（包括药物治疗）都至关重要递送、激活调节和抗原加工。还不够的是了解受影响的特定吞噬体-内体过程或病原体利用的生理和生化机制合理设计干预措施以恢复 PC 功能。主要目标是这个提议是为了确定哪些生理变化发生在吞噬体-内体过程导致这些抗微生物药物失效 IV 与充分表征的相互作用期间的细胞活动吞噬细胞J774.1鼠巨噬细胞。巨噬细胞信息- 病原体的影响落后于其他电脑，但在免疫抑制它们可能成为剩余的主要PC。 IV 是一个重要的疾病及其对 PC 的许多影响与细菌，但它更简单，因此识别的前景重要的 PC 抑制剂机制和改进抗病毒疗法是增加。具体目标是： 1) 确定影响的范围和类型暴露后巨噬细胞胞内运输途径的变化四． 2）确定细胞内物理化学发生什么变化早期巨噬细胞和细胞内区室的环境流感病毒暴露。 3) 研究药剂的效果模拟或拮抗 IV 引起的巨噬细胞变化内体和吞噬体功能。定量数字视频显微镜将用于检查 IV 对 J774 细胞内囊泡的影响基于运输功能，包括之间的吸收和移动特定内吞和吞噬的分子标记区室功能。荧光探针也将用于监测物理- 吞噬体和内体的化学状态。预计双方方法学和细胞生物学发现将适用于了解其他病毒类型、细菌和寄生虫对 PC 的抑制作用。