Drosophila melanogaster S2 cells:: a model system to study Chlamydia interaction with host cells
Drosophila melanogaster S2 cells:: a model system to study Chlamydia interaction with host cells
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DOI：
10.1111/j.1462-5822.2005.00508.x
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发表时间：
2005-05-01
影响因子：
3.4
通讯作者：
Engel, JN
中科院分区：
文献类型：
--
作者：
Elwell, C;Engel, JN
衣原体属物种是重要的人类疾病的主要原因，但解剖宿主-病原体相互作用受到缺乏细菌遗传学和难以在哺乳动物宿主中进行正向遗传筛选的阻碍。基于RNA干扰（RNAi）的基因失活方法现在可以很容易地在遗传上易于处理的模型宿主，如果蝇，并提供了一种新的方法来确定宿主基因的发病所需的。我们检测了沙眼衣原体感染D。黑腹S2细胞再现了哺乳动物细胞感染的关键方面。在哺乳动物细胞中，C.肝素和细胞松弛素D可显著减少沙眼衣原体的侵入。包涵体在S2细胞中形成，获得高尔基体衍生的鞘脂，并避免吞噬溶酶体融合。观察到初级体（EB）向网状体（RB）分化，但未观察到RB向EB的发育或宿主细胞杀伤。RNAi介导的Rac失活，Rac是一种Rho GT酶，最近被证明是C.沙眼衣原体进入哺乳动物细胞，抑制C.沙眼衣原体感染的S2细胞。我们的结论是果蝇S2细胞忠实地模仿衣原体宿主细胞相互作用的早期事件，并提供了一个真正的系统，系统地解剖宿主功能的重要致病专性细胞内病原体。
Chlamydia spp. are major causes of important human diseases, but dissecting the host-pathogen interactions has been hampered by the lack of bacterial genetics and the difficulty in carrying out forward genetic screens in mammalian hosts. RNA interference (RNAi)-based methodologies for gene inactivation can now be easily carried out in genetically tractable model hosts, such as Drosophila melanogaster, and offer a new approach to identifying host genes required for pathogenesis. We tested whether Chlamydia trachomatis infection of D. melanogaster S2 cells recapitulated critical aspects of mammalian cell infections. As in mammalian cells, C. trachomatis entry was greatly reduced by heparin and cytochalasin D. Inclusions were formed in S2 cells, acquired Golgi-derived sphingolipids, and avoided phagolysosomal fusion. Elementary body (EB) to reticulate body (RB) differentiation was observed, however, no RB to EB development or host cell killing was observed. RNAi-mediated inactivation of Rac, a Rho GTPase recently shown to be required for C. trachomatis entry in mammalian cells, inhibits C. trachomatis infection in S2 cells. We conclude that Drosophila S2 cells faithfully mimic early events in Chlamydia host cell interactions and provides a bona fide system to systematically dissect host functions important in the pathogenesis of obligate intracellular pathogens.