Listeriolysin O-dependent activation of endothelial cells during infection with Listeria monocytogenes:: activation of NF-κB and upregulation of adhesion molecules and chemokines
Listeriolysin O-dependent activation of endothelial cells during infection with Listeria monocytogenes:: activation of NF-κB and upregulation of adhesion molecules and chemokines
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DOI：
10.1046/j.1365-2958.1999.01305.x
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发表时间：
1999-03-01
影响因子：
3.6
通讯作者：
Berche, P
中科院分区：
文献类型：
--
作者：
Kayal, S;Lilienbaum, A;Berche, P
兼性细胞内细菌单核细胞增生李斯特菌是一种侵入性病原体，其穿过血管内皮并传播到胎盘和中枢神经系统。它与内皮细胞的相互作用是至关重要的发病机制，动脉粥样硬化。用L.在单核细胞增多症中，我们发现野生型细菌诱导粘附分子（ICAM-1和E-选择素）的表达、趋化因子分泌（IL-8和单核细胞趋化蛋白-1）和NF-κ B B核转位。HUVEC的激活需要活菌，而在L.单核细胞增多症，表明毒力基因与内皮细胞活化有关。使用遗传方法与突变体的毒力基因，我们发现，溶血素O（LLO）缺陷突变体失活的hly基因不诱导HUVEC激活，而不是突变体失活的其他毒力基因。粘附分子表达、趋化因子分泌和NF-κ B活化被用编码LLO的hly基因转化的无害李斯特菌菌株完全恢复。在携带NF-κ B反应性lacZ报告基因的转基因小鼠中研究了内皮细胞活化与病毒发病机制的体内相关性，通过在感染有毒性溶血性菌株的小鼠的毛细血管内皮细胞中强烈的lacZ表达来观察NF-κ B B活化，但在感染有非溶血性同基因突变体的小鼠中没有观察到。通过静脉内注射纯化的LLO，从而诱导毛细血管内皮细胞中NF-κ B的刺激，提供了LLO参与转基因小鼠中NF-κ B活化的直接证据。我们的研究结果表明，细菌分泌的功能性溶血素O有助于作为一种有效的炎症刺激，诱导内皮细胞活化在感染过程中。
The facultative intracellular bacterium Listeria monocytogenes is an invasive pathogen that crosses the vascular endothelium and disseminates to the placenta and the central nervous system. Its interaction with endothelial cells is crucial for the pathogenesis of listeriosis. By infecting in vitro human umbilical vein endothelial cells (HUVEC) with L. monocytogenes, we found that wild-type bacteria induced the expression of the adhesion molecules (ICAM-1 and E-selectin), chemokine secretion (IL-8 and monocyte chemotactic protein-1) and NF-kappa B nuclear translocation. The activation of HUVEC required viable bacteria and was abolished in prfA-deficient mutants of L. monocytogenes, suggesting that virulence genes are associated with endothelial cell activation. Using a genetic approach with mutants of virulence genes, we found that listeriolysin O (LLO)-deficient mutants inactivated in the hly gene did not induce HUVEC activation, as opposed to mutants inactivated in the other virulence genes. Adhesion molecule expression, chemokine secretion and NF-kappa B activation were fully restored by a strain of Listeria innocua transformed with the hly gene encoding LLO. The relevance in vivo of endothelial cell activation for listerial pathogenesis was investigated in transgenic mice carrying an NF-kappa B-responsive lacZ reporter gene, NF-kappa B activation was visualized by a strong lacZ expression in endothelial cells of capillaries of mice infected with a virulent haemolytic strain, but was not seen in those infected with a non-haemolytic isogenic mutant. Direct evidence that LLO is involved in NF-kappa B activation in transgenic mice was provided by injecting intravenously purified LLO, thus inducing stimulation of NF-kappa B in endothelial cells of blood capillaries. Our results demonstrate that functional listeriolysin O secreted by bacteria contributes as a potent inflammatory stimulus to inducing endothelial cell activation during the infectious process.