title: Antitumor immunity triggered by melphalan is potentiated by melanoma cell surface-associated calreticulin.
abstract: Systemic chemotherapy generally has been considered immunosuppressive, but it has become evident that certain chemotherapeutic drugs elicit immunogenic danger signals in dying cancer cells that can incite protective antitumor immunity. In this study, we investigated whether locoregionally applied therapies, such as melphalan, used in limb perfusion for melanoma (Mel-ILP) produce related immunogenic effects. In human melanoma biopsies, Mel-ILP treatment upregulated IL1B, IL8, and IL6 associated with their release in patients' locoregional sera. Although induction of apoptosis in melanoma cells by melphalan in vitro did not elicit threshold levels of endoplasmic reticulum and reactive oxygen species stress associated with danger signals, such as induction of cell-surface calreticulin, prophylactic immunization and T-cell depletion experiments showed that melphalan administration in vivo could stimulate a CD8(+) T cell-dependent protective antitumor response. Interestingly, the vaccination effect was potentiated in combination with exogenous calreticulin, but not tumor necrosis factor, a cytokine often combined with Mel-ILP. Our results illustrate how melphalan triggers inflammatory cell death that can be leveraged by immunomodulators such as the danger signal calreticulin.
authors.affiliation: Cell Death Research and Therapy Laboratory, Department of Cellular and Molecular Medicine, Faculty of Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Aleksandra M
authors.initials: AM
authors.lastname: Dudek-Perić
authors.affiliation: Laboratory of Clinical and Experimental Endocrinology, Department of Clinical and Experimental Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Gabriela B
authors.initials: GB
authors.lastname: Ferreira
authors.affiliation: Department of Immunology, Center of Biostructure Research, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Angelika
authors.initials: A
authors.lastname: Muchowicz
authors.affiliation: Translational Cell and Tissue Research, Department of Imaging and Pathology, Faculty of Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Jasper
authors.initials: J
authors.lastname: Wouters
authors.affiliation: Antiviral Immunity, Biotherapy and Vaccine Unit, Infection and Epidemiology Department, Institute Pasteur, Paris, France.
authors.forename: Nicole
authors.initials: N
authors.lastname: Prada
authors.affiliation: Cell Death Research and Therapy Laboratory, Department of Cellular and Molecular Medicine, Faculty of Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Shaun
authors.initials: S
authors.lastname: Martin
authors.affiliation: Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Faculty of Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Santeri
authors.initials: S
authors.lastname: Kiviluoto
authors.affiliation: Department of Immunology, Center of Biostructure Research, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Winiarska
authors.affiliation: Bioceros, CM Utrecht, the Netherlands.
authors.forename: Louis
authors.initials: L
authors.lastname: Boon
authors.affiliation: Laboratory of Clinical and Experimental Endocrinology, Department of Clinical and Experimental Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Chantal
authors.initials: C
authors.lastname: Mathieu
authors.affiliation: Translational Cell and Tissue Research, Department of Imaging and Pathology, Faculty of Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Joost
authors.initials: J
authors.lastname: van den Oord
authors.affiliation: Surgical Oncology, Department of Oncology, KU Leuven, Leuven, Belgium.
authors.forename: Marguerite
authors.initials: M
authors.lastname: Stas
authors.affiliation: Antiviral Immunity, Biotherapy and Vaccine Unit, Infection and Epidemiology Department, Institute Pasteur, Paris, France.
authors.forename: Marie-Lise
authors.initials: ML
authors.lastname: Gougeon
authors.affiliation: Department of Immunology, Center of Biostructure Research, Medical University of Warsaw, Warsaw, Poland. Institute of Physical Chemistry, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: Jakub
authors.initials: J
authors.lastname: Golab
authors.affiliation: Cell Death Research and Therapy Laboratory, Department of Cellular and Molecular Medicine, Faculty of Medicine, KU Leuven, Leuven, Belgium. Patrizia.Agostinis@med.kuleuven.be abhishek.garg@med.kuleuven.be.
authors.forename: Abhishek D
authors.initials: AD
authors.lastname: Garg
authors.affiliation: Cell Death Research and Therapy Laboratory, Department of Cellular and Molecular Medicine, Faculty of Medicine, KU Leuven, Leuven, Belgium. Patrizia.Agostinis@med.kuleuven.be abhishek.garg@med.kuleuven.be.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Agostinis
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000502:physiology*; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D037282:Calreticulin / Q000502:physiology*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects / Q000502:physiology; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D008545:Melanoma / Q000188:drug therapy / Q000276:immunology*; D008558:Melphalan / Q000494:pharmacology*; D051379:Mice; D017382:Reactive Oxygen Species / Q000378:metabolism / Q000494:pharmacology; D012878:Skin Neoplasms / Q000188:drug therapy / Q000276:immunology*
chemical_list: D000954:Antigens, Surface; D018906:Antineoplastic Agents, Alkylating; D037282:Calreticulin; D016207:Cytokines; D018836:Inflammation Mediators; D017382:Reactive Oxygen Species; D008558:Melphalan