Killing Bacteria with Cytotoxic Effector Proteins of Human Killer Immune Cells: Granzymes, Granulysin, and Perforin
Killing Bacteria with Cytotoxic Effector Proteins of Human Killer Immune Cells: Granzymes, Granulysin, and Perforin
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DOI：
10.1007/978-1-4939-6673-8_18
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发表时间：
2017-01-01
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影响因子：
--
通讯作者：
Walch, Michael
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文献类型：
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作者：
Leon, Diego Lopez;Fellay, Isabelle;Walch, Michael
细菌性病原体对人类健康构成持续威胁，近年来，对最后手段抗生素产生耐药性的菌株急剧增加，加剧了这一威胁。高等脊椎动物的免疫系统通常进化出几种有效的先天和适应性机制来对抗无处不在的细菌病原体。在这些机制中，免疫蛋白酶被认为在抗菌免疫防御中起着重要作用。适应性免疫系统的效应丝氨酸蛋白酶，即颗粒酶，当它们被原核膜破坏蛋白（如颗粒酶）递送到细菌细胞质中时，会发挥强大的抗菌活性。在本章中，我们详细介绍了在体外研究人类颗粒酶和颗粒酶对细胞外和细胞内细菌病原体的协同细胞毒性作用的实验方案。此外，我们提供了一种简单而快速的方法来生化纯化天然细胞毒性效应分子，这是进行此类研究所必需的。
Bacterial pathogens represent a constant threat to human health that was exacerbated in recent years by a dramatic increase of strains resistant to last resort antibiotics. The immune system of higher vertebrates generally evolved several efficient innate and adaptive mechanisms to fight ubiquitous bacterial pathogens. Among those mechanisms, immune proteases were recognized to contribute essentially to antibacterial immune defense. The effector serine proteases of the adaptive immune system, the granzymes, exert potent antimicrobial activity when they are delivered into the bacterial cytosol by prokaryotic membrane disrupting proteins, such as granulysin.In this chapter, we are detailing experimental protocols to study the synergistic cytotoxic effects of human granzymes and granulysin on extracellular as well as on intracellular bacterial pathogens in vitro. In addition, we provide a simple and fast-forward method to biochemically purify native cytotoxic effector molecules necessary to perform this kind of investigations.