Abstract
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.
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Acknowledgments
This work was supported by grants from the Novartis Foundation for Medical-Biological Research, from the Research Pool and Fund of the University of Fribourg, as well as from the Gottfried and Julia Bangerter-Rhyner foundation (to M.W. and P.Y.M.). We thank Luis Filgueira at the University of Fribourg and Denis Martinvalet at the University of Geneva for helpful discussions. We also acknowledge the contributions of Judy Lieberman and Farokh Dotiwala at Harvard Medical School to the design and establishment of the outlined methods.
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León, D.L., Fellay, I., Mantel, PY., Walch, M. (2017). Killing Bacteria with Cytotoxic Effector Proteins of Human Killer Immune Cells: Granzymes, Granulysin, and Perforin. In: Nordenfelt, P., Collin, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology, vol 1535. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6673-8_18
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DOI: https://doi.org/10.1007/978-1-4939-6673-8_18
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