title: Impaired killing of Candida albicans by granulocytes mobilized for transfusion purposes: a role for granule components.
abstract: Granulocyte transfusions are used to treat neutropenic patients with life-threatening bacterial or fungal infections that do not respond to anti-microbial drugs. Donor neutrophils that have been mobilized with granulocyte-colony stimulating factor (G-CSF) and dexamethasone are functional in terms of antibacterial activity, but less is known about their fungal killing capacity. We investigated the neutrophil-mediated cytotoxic response against C. albicans and A. fumigatus in detail. Whereas G-CSF/dexamethasone-mobilized neutrophils appeared less mature as compared to neutrophils from untreated controls, these cells exhibited normal ROS production by the NADPH oxidase system and an unaltered granule mobilization capacity upon stimulation. G-CSF/dexamethasone-mobilized neutrophils efficiently inhibited A. fumigatus germination and killed Aspergillus and Candida hyphae, but the killing of C. albicans yeasts was distinctly impaired. Following normal Candida phagocytosis, analysis by mass spectrometry of purified phagosomes after fusion with granules demonstrated that major constituents of the antimicrobial granule components, including major basic protein (MBP), were reduced. Purified MBP showed candidacidal activity, and neutrophil-like Crisp-Cas9 NB4-KO-MBP differentiated into phagocytes were impaired in Candida killing. Together, these findings indicate that G-CSF/dexamethasone-mobilized neutrophils for transfusion purposes have a selectively impaired capacity to kill Candida yeasts, as a consequence of an altered neutrophil granular content.
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands r.gazendam@sanquin.nl.
authors.forename: Roel P
authors.initials: RP
authors.lastname: Gazendam
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Annemarie
authors.initials: A
authors.lastname: van de Geer
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: John L
authors.initials: JL
authors.lastname: van Hamme
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Anton T J
authors.initials: AT
authors.lastname: Tool
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Dieke J
authors.initials: DJ
authors.lastname: van Rees
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Cathelijn E M
authors.initials: CE
authors.lastname: Aarts
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Maartje
authors.initials: M
authors.lastname: van den Biggelaar
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Floris
authors.initials: F
authors.lastname: van Alphen
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Paul
authors.initials: P
authors.lastname: Verkuijlen
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Alexander B
authors.initials: AB
authors.lastname: Meijer
authors.affiliation: The Netherlands Netherlands Cancer Institute, Division of Cell Biology, Amsterdam, The Netherlands.
authors.forename: Hans
authors.initials: H
authors.lastname: Janssen
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Dirk
authors.initials: D
authors.lastname: Roos
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Timo K
authors.initials: TK
authors.lastname: van den Berg
authors.affiliation: Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands Emma Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Taco W
authors.initials: TW
authors.lastname: Kuijpers
mesh_terms: D015415:Biomarkers; D002176:Candida albicans / Q000276:immunology*; D015550:Cell Degranulation / Q000187:drug effects / Q000276:immunology; D003594:Cytoplasmic Granules / Q000276:immunology / Q000378:metabolism; D003602:Cytotoxicity, Immunologic; D003907:Dexamethasone / Q000494:pharmacology; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006098:Granulocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D006801:Humans; D016130:Immunophenotyping; D017708:Leukocyte Transfusion; D050296:Microbial Viability / Q000276:immunology*; D019255:NADPH Oxidases / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D010587:Phagocytosis / Q000187:drug effects / Q000276:immunology; D010588:Phagosomes / Q000276:immunology / Q000382:microbiology
chemical_list: D015415:Biomarkers; D016179:Granulocyte Colony-Stimulating Factor; D003907:Dexamethasone; D019255:NADPH Oxidases