title: Killing Bacteria with Cytotoxic Effector Proteins of Human Killer Immune Cells: Granzymes, Granulysin, and Perforin.
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.
authors.affiliation: Unit of Anatomy, Department of Medicine, University of Fribourg, Rue A.-Gockel 1, 1700, Fribourg, Switzerland.
authors.forename: Diego López
authors.initials: DL
authors.lastname: León
authors.affiliation: Unit of Anatomy, Department of Medicine, University of Fribourg, Rue A.-Gockel 1, 1700, Fribourg, Switzerland.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Fellay
authors.affiliation: Unit of Anatomy, Department of Medicine, University of Fribourg, Rue A.-Gockel 1, 1700, Fribourg, Switzerland.
authors.forename: Pierre-Yves
authors.initials: PY
authors.lastname: Mantel
authors.affiliation: Unit of Anatomy, Department of Medicine, University of Fribourg, Rue A.-Gockel 1, 1700, Fribourg, Switzerland. michael.walch@unifr.ch.
authors.forename: Michael
authors.initials: M
authors.lastname: Walch
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects* / Q000276:immunology*; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D004305:Dose-Response Relationship, Drug; D005110:Extracellular Space / Q000276:immunology / Q000382:microbiology; D053804:Granzymes / Q000494:pharmacology*; D006801:Humans; D042541:Intracellular Space / Q000276:immunology / Q000382:microbiology; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism; D054353:Perforin / Q000494:pharmacology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D000945:Antigens, Differentiation, T-Lymphocyte; C075005:GNLY protein, human; D054353:Perforin; D053804:Granzymes
keywords: Antibacterial activity; Cell-mediated cytotoxicity; Granulysin; Granzymes; Immune serine proteases