Apidianakis Yiorgos, Rahme Laurence G
Department of Surgery, Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA.
Nat Protoc. 2009;4(9):1285-94. doi: 10.1038/nprot.2009.124. Epub 2009 Aug 13.
Conservation of host signaling pathways and tissue physiology between Drosophila melanogaster and mammals allows for the modeling of human host-pathogen interactions in Drosophila. Here we present the use of genetically tractable Drosophila models of bacterial pathogenesis to study infection with the human opportunistic pathogen Pseudomonas aeruginosa. We describe and compare two protocols commonly used to infect Drosophila with P. aeruginosa: needle-pricking and injector-pumping. Each model has relevance for examining host components and bacterial factors in host defense and virulence. Fly survival and bacterial proliferation within host flies can be assessed as a measure of host susceptibility and pathogen virulence potential. The profiles of host responses toward P. aeruginosa virulent and non-virulent strains can be determined, enabling the identification of interaction-specific genes that could potentially favor or limit the initiation and progression of infection. Both of the protocols presented herein may be adapted for the inoculation and study of other microbial pathogens. P. aeruginosa cell preparation requires 24 h, fly inoculation 1 h, and fly survival and bacterial proliferation 1-4 d.
黑腹果蝇与哺乳动物之间宿主信号通路和组织生理学的保守性使得在果蝇中模拟人类宿主与病原体的相互作用成为可能。在此,我们展示了利用遗传易处理的果蝇细菌致病模型来研究人类机会致病菌铜绿假单胞菌的感染。我们描述并比较了两种常用于用铜绿假单胞菌感染果蝇的方案:针刺法和注射器注射法。每个模型对于研究宿主防御和毒力中的宿主成分及细菌因素都具有相关性。果蝇的存活率以及宿主果蝇体内细菌的增殖情况可作为衡量宿主易感性和病原体毒力潜力的指标进行评估。可以确定宿主对铜绿假单胞菌有毒和无毒菌株的反应谱,从而能够鉴定出可能有利于或限制感染起始和进展的相互作用特异性基因。本文介绍的两种方案均可适用于其他微生物病原体的接种和研究。铜绿假单胞菌细胞制备需要24小时,果蝇接种需要1小时,果蝇存活和细菌增殖需要1 - 4天。