Drosophila S2 cells:: an alternative infection model for Listeria monocytogenes
Drosophila S2 cells:: an alternative infection model for Listeria monocytogenes
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DOI：
10.1046/j.1462-5822.2003.00327.x
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发表时间：
2003-12-01
影响因子：
3.4
通讯作者：
Portnoy, DA
中科院分区：
文献类型：
--
作者：
Cheng, LW;Portnoy, DA
单核细胞增生李斯特菌是一种革兰氏阳性兼性胞内细菌病原体，感染人类和动物。其致病策略涉及介导胞内生长和细胞间传播的毒力蛋白的表达。一个关键的毒力蛋白是胆固醇依赖性溶细胞素，溶细胞素O（LLO），它主要负责介导从吞噬体逃逸到宿主细胞溶质中。为了进一步研究L.为了研究单核细胞增多症感染，我们试图开发果蝇S2细胞作为感染模型。在这里，我们表明，S2细胞与感染的哺乳动物细胞培养模型共享许多特性。与小鼠巨噬细胞一样，吞噬体从S2细胞逃逸需要LLO。此外，液泡逃逸依赖于它们通过ATP酶质子泵的酸化，因为巴弗洛霉素A（1）处理急剧减少逃逸。然而，与小鼠巨噬细胞不同，LLO突变体在S2细胞的吞噬体中复制。果蝇细胞是胆固醇营养缺陷型细胞，外源胆固醇增加了L.单核细胞增多症（LLO独立的），也增加了空泡逃逸的效率（LLO依赖的）。利用RNA干扰等遗传学工具，S2细胞可能成为研究宿主-病原体相互作用的重要模型。
Listeria monocytogenes is a Gram-positive facultative intracellular bacterial pathogen that infects humans and animals. Its pathogenic strategy involves the expression of virulence proteins that mediate intracytosolic growth and cell-to-cell spread. A key virulence protein is the cholesterol-dependent cytolysin, listeriolysin O (LLO), which is largely responsible for mediating escape from the phagosome into the host cytosol. To study further the host processes exploited during L. monocytogenes infection, we sought to develop Drosophila S2 cells as a model for infection. Here, we show that S2 cells share a number of properties with mammalian cell culture models of infection. As with mouse macrophages, LLO was required for phagosomal escape from S2 cells. Furthermore, vacuolar escape was dependent on their acidification via the ATPase proton pumps, as bafilomycin A(1) treatment sharply decreased escape. However, unlike in mouse macrophages, LLO mutants replicated in the phagosome of S2 cells. Drosophila cells are cholesterol auxotrophs, and exogenous cholesterol increased the infection rate of L. monocytogenes (LLO independent) and also augmented the efficiency of vacuolar escape (LLO dependent). With available genetic tools such as RNA interference, S2 cells could become an important model in the study of host-pathogen interactions.