Real-time studies of the progression of bacterial infections and immediate tissue responses in live animals
Real-time studies of the progression of bacterial infections and immediate tissue responses in live animals
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DOI：
10.1111/j.1462-5822.2006.00799.x
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发表时间：
2007-02-01
影响因子：
3.4
通讯作者：
Richter-Dahlfors, Agneta
中科院分区：
文献类型：
--
作者：
Mansson, Lisa E.;Melican, Keira;Richter-Dahlfors, Agneta
通过结合活体多光子显微镜和细菌遗传学，我们开发了一种能够对活体动物中细菌增殖和组织反应进行实时成像的技术。通过将表达 GFP(+) 的尿路致病性大肠杆菌 (UPEC) 显微注射到活体大鼠外置肾小管中，实现了感染过程的空间和时间控制。 GFP(+)作为单拷贝染色体基因融合体被引入临床UPEC菌株CFT073中。数小时内，细菌定植伴随着明显的缺血效应、血管周围渗漏、肾小管完整性丧失和免疫细胞局部募集。缺乏外毒素溶血素的同基因细菌菌株的病理生理学发生了改变，揭示了体内细菌毒力因子的微妙和暂时的作用。对注射的肾单位进行显微解剖和 RNA 提取，可以对原核和真核基因表达进行分子分析。这里描述的技术可用于研究多种细菌病原体引起的整合细胞通讯，有助于设计对抗细菌感染的策略。
By combining intravital multiphoton microscopy and bacterial genetics we have developed a technique enabling real-time imaging of bacterial proliferation and tissue responses in a live animal. Spatial and temporal control of the infection process was achieved by microinjecting GFP(+)-expressing uropathogenic Escherichia coli (UPEC) into tubules of exteriorized kidneys in live rats. GFP(+) was introduced in the clinical UPEC strain CFT073 as a single-copy chromosomal gene fusion. Within hours, bacterial colonization was accompanied by marked ischaemic effects, perivascular leakage, loss of tubular integrity and localized recruitment of immune cells. The pathophysiology was altered in response to an isogenic bacterial strain lacking the exotoxin haemolysin, revealing the subtle and temporal roles of bacterial virulence factors in vivo. Microdissection and RNA extraction of the injected nephron allowed molecular analysis of prokaryotic and eukaryotic gene expression. The techniques described here can be applied to study the integrated cell communication evoked by a variety of bacterial pathogens, assisting in the design of strategies to combat bacterial infections.