Real-time high resolution 3D imaging of the lyme disease spirochete adhering to and escaping from the vasculature of a living host.
Real-time high resolution 3D imaging of the lyme disease spirochete adhering to and escaping from the vasculature of a living host.
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DOI：
10.1371/journal.ppat.1000090
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发表时间：
2008-06-20
期刊：
影响因子：
6.7
通讯作者：
Chaconas G
中科院分区：
文献类型：
--
作者：
Moriarty TJ;Norman MU;Colarusso P;Bankhead T;Kubes P;Chaconas G
致病性螺旋体是在世界范围内引起许多新出现和再出现疾病的细菌，包括梅毒、钩端螺旋体病、回归热和莱姆病。它们通过致密的细胞外基质有效地导航，并以未知的机制穿过血脑屏障。由于其细长的形态，螺旋体很难通过标准光学显微镜可视化，阻碍了对其原位行为的研究。我们设计了一种表达绿色荧光蛋白（GFP）的莱姆病病原体伯氏疏螺旋体的荧光感染菌株。活体小鼠微血管荧光螺旋体播散的实时3D和4D高分辨率定量分析显示，螺旋体播散是一个多阶段的过程，包括短暂的系链型关联、短期的拖拽相互作用和固定粘附。固定黏附和外渗螺旋体最常见于内皮连接处，螺旋体的平移运动似乎在跨内皮迁移中起着不可或缺的作用。据我们所知，这是首次报道细菌病原体在活体哺乳动物宿主中传播的高分辨率3D和4D可视化，并首次直接了解螺旋体在体内的传播。致病性螺旋体是在世界范围内引起许多新出现和再出现疾病的细菌，包括梅毒、钩端螺旋体病、回归热和莱姆病。它们表现出一种不寻常的运动形式，可以感染许多不同的组织；然而，它们从血液传播到靶部位的机制尚不清楚。在活体宿主的单细胞水平上直接可视化细菌病原体是微生物学的一个重要目标，因为这种方法可能对疾病过程产生关键的见解。我们设计了一种莱姆病病原体伯氏疏螺旋体（Borrelia burgdorferi）的荧光菌株，并使用传统和旋转盘共聚焦活体显微镜在活体小鼠中实时和3D地直接观察这些细菌。我们发现螺旋体与脉管系统的相互作用和传播是一个意想不到的复杂的多阶段过程，螺旋体的运动似乎在传播中起着不可或缺的作用。据我们所知，这是第一份关于细菌病原体在活体哺乳动物宿主中传播的高分辨率3D可视化报告，并首次提供了对活体螺旋体传播的直接见解。
Pathogenic spirochetes are bacteria that cause a number of emerging and re-emerging diseases worldwide, including syphilis, leptospirosis, relapsing fever, and Lyme borreliosis. They navigate efficiently through dense extracellular matrix and cross the blood–brain barrier by unknown mechanisms. Due to their slender morphology, spirochetes are difficult to visualize by standard light microscopy, impeding studies of their behavior in situ. We engineered a fluorescent infectious strain of Borrelia burgdorferi, the Lyme disease pathogen, which expressed green fluorescent protein (GFP). Real-time 3D and 4D quantitative analysis of fluorescent spirochete dissemination from the microvasculature of living mice at high resolution revealed that dissemination was a multi-stage process that included transient tethering-type associations, short-term dragging interactions, and stationary adhesion. Stationary adhesions and extravasating spirochetes were most commonly observed at endothelial junctions, and translational motility of spirochetes appeared to play an integral role in transendothelial migration. To our knowledge, this is the first report of high resolution 3D and 4D visualization of dissemination of a bacterial pathogen in a living mammalian host, and provides the first direct insight into spirochete dissemination in vivo. Pathogenic spirochetes are bacteria that cause a number of emerging and re-emerging diseases worldwide, including syphilis, leptospirosis, relapsing fever, and Lyme disease. They exhibit an unusual form of motility and can infect many different tissues; however, the mechanism by which they disseminate from the blood to target sites is unknown. Direct visualization of bacterial pathogens at the single cell level in living hosts is an important goal of microbiology, since this approach is likely to yield critical insight into disease processes. We engineered a fluorescent strain of Borrelia burgdorferi, a Lyme disease pathogen, and used conventional and spinning disk confocal intravital microscopy to directly visualize these bacteria in real time and 3D in living mice. We found that spirochete interaction with and dissemination out of the vasculature was a multi-stage process of unexpected complexity and that spirochete movement appeared to play an integral role in dissemination. To our knowledge, this is the first report of high resolution 3D visualization of dissemination of a bacterial pathogen in a living mammalian host, and provides the first direct insight into spirochete dissemination in vivo.
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影响因子：
3.6
作者：
Eggers, CH;Caimano, MJ;Radolf, JD
通讯作者：
Radolf, JD
影响因子：
3.8
作者：
COMSTOCK, LE;THOMAS, DD
通讯作者：
THOMAS, DD
影响因子：
7.8
作者：
Albelda, S M;Muller, W A;Buck, C A;Newman, P J
通讯作者：
Newman, P J
DOI：
10.1073/pnas.91.8.3433
发表时间：
1994-04-12
影响因子：
11.1
作者：
GOLDSTEIN, SF;CHARON, NW;KREILING, JA
通讯作者：
KREILING, JA
影响因子：
3.1
作者：
Coburn, J;Magoun, L;Leong, JM
通讯作者：
Leong, JM