Methods to monitor autophagy in H. pylori vacuolating cytotoxin A (VacA)-treated cells
Methods to monitor autophagy in H. pylori vacuolating cytotoxin A (VacA)-treated cells
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DOI：
10.4161/auto.6.1.10222
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发表时间：
2010-01
期刊：
影响因子：
13.3
通讯作者：
Deepa Raju;N. Jones
中科院分区：
文献类型：
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作者：
Deepa Raju;N. Jones
幽门螺杆菌是一种革兰氏阴性病原体，感染世界上至少一半的人口，不仅与胃癌有关，还与胃炎和消化性溃疡等其他疾病有关。事实上，幽门螺杆菌被认为是胃癌发展的最重要的单一危险因素。幽门螺杆菌分泌的空泡细胞毒素 VacA 可促进细菌的细胞内存活并调节宿主免疫反应。在最近的一项研究中，我们报道了 VacA 诱导自噬。在胃上皮细胞中检测到多层自噬体，与 VacA 形成的大液泡不同。此外，抑制自噬可以稳定 VacA 并减少细胞中的空泡形成，表明毒素正在被自噬降解，从而限制毒素诱导的宿主细胞损伤。本研究中使用的许多方法都是适用于幽门螺杆菌感染和 VacA 中毒的常用技术。在本文中，我们描述了用于评估和监测幽门螺杆菌感染期间自噬的各种方法和具体方案。
Helicobacter pylori is a gram negative pathogen that infects at least half of the world’s population and is associated not only with gastric cancer but also with other diseases such as gastritis and peptic ulcers. Indeed, H. pylori is considered the single most important risk factor for the development of gastric cancer. The vacuolating cytotoxin, VacA, secreted by H. pylori promotes intracellular survival of the bacterium and modulates host immune responses. In a recent study, we reported that VacA induces autophagy. Multilamellar autophagosomes are detected in gastric epithelial cells that are distinct from the large vacuoles formed by VacA. Furthermore, inhibition of autophagy stabilizes VacA and reduces vacuolation in the cells indicating that the toxin is being degraded by autophagy, thus limiting toxin-induced host cell damage. Many of the methods that were used for this study are commonly employed techniques that were adapted for H. pylori infection and VacA intoxication. In this paper, we describe the various methods and specific protocols used for the assessment and monitoring of autophagy during H. pylori infection.