Infection of zebrafish embryos with live fluorescent Streptococcus pneumoniae as a real-time pneumococcal meningitis model
Infection of zebrafish embryos with live fluorescent Streptococcus pneumoniae as a real-time pneumococcal meningitis model
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DOI：
10.1186/s12974-016-0655-y
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发表时间：
2016-08-19
影响因子：
9.3
通讯作者：
Vandenbroucke-Grauls, Christina M. J. E.
中科院分区：
文献类型：
--
作者：
Jim, Kin Ki;Engelen-Lee, JooYeon;Vandenbroucke-Grauls, Christina M. J. E.
背景：肺炎链球菌是细菌性脑膜炎最重要的病因之一，细菌性脑膜炎是一种由细菌和宿主来源的毒素导致不良后果的感染。在这项研究中，我们开发并表征了斑马鱼胚胎中的肺炎球菌脑膜炎模型，该模型允许实时调查早期宿主-微生物相互作用。方法：用绿色荧光野生型5感染斑马鱼尾静脉或后脑室胚胎。肺炎球菌D39或溶血素缺陷突变体。用kdrl:mCherry转基因斑马鱼细胞系观察血管，用远红色抗l -plastin或mpx:GFP/mpeg1:mCherry斑马鱼染色观察吞噬细胞，其中有绿色荧光中性粒细胞和红色荧光巨噬细胞。通过荧光共聚焦和延时显微镜进行成像。结果：尾静脉感染后，可见肺炎球菌在血管内的局灶性堵塞，细菌通过血脑屏障迁移到蛛网膜下腔和脑组织。与野生型菌株相比，后脑室肺炎溶素缺陷链球菌的生长和迁移减弱。延时和共聚焦成像显示，最初的先天免疫反应5。蛛网膜下腔内的肺炎主要由中性粒细胞组成，溶肺素介导的细胞溶解活性导致吞噬细胞明显减少。结论：这种新的脑膜炎模型可以实时详细分析和可视化肺炎球菌脑膜炎中宿主-微生物相互作用，是一种非常有前途的工具，可以进一步深入了解肺炎球菌脑膜炎的发病机制。
Background: Streptococcus pneumoniae is one of the most important causes of bacterial meningitis, an infection where unfavourable outcome is driven by bacterial and host-derived toxins. In this study, we developed and characterized a pneumococcal meningitis model in zebrafish embryos that allows for real-time investigation of early host-microbe interaction.Methods: Zebrafish embryos were infected in the caudal vein or hindbrain ventricle with green fluorescent wild-type 5. pneumoniae D39 or a pneumolysin-deficient mutant. The kdrl:mCherry transgenic zebrafish line was used to visualize the blood vessels, whereas phagocytic cells were visualized by staining with far red anti-L-plastin or in mpx:GFP/mpeg1:mCherry zebrafish, that have green fluorescent neutrophils and red fluorescent macrophages. Imaging was performed by fluorescence confocal and time-lapse microscopy.Results: After infection by caudal vein, we saw focal clogging of the pneumococci in the blood vessels and migration of bacteria through the blood-brain barrier into the subarachnoid space and brain tissue. Infection with pneumolysin-deficient S. pneumoniae in the hindbrain ventricle showed attenuated growth and migration through the brain as compared to the wild-type strain. Time-lapse and confocal imaging revealed that the initial innate immune response to 5. pneumonie in the subarachnoid space mainly consisted of neutrophils and that pneumolysin-mediated cytolytic activity caused a marked reduction of phagocytes.Conclusions: This new meningitis model permits detailed analysis and visualization of host-microbe interaction in pneumococcal meningitis in real time and is a very promising tool to further our insights in the pathogenesis of pneumococcal meningitis.