title: Zinc and Manganese Chelation by Neutrophil S100A8/A9 (Calprotectin) Limits Extracellular Aspergillus fumigatus Hyphal Growth and Corneal Infection.
abstract: Calprotectin, a heterodimer of S100A8 and S100A9, is an abundant neutrophil protein that possesses antimicrobial activity primarily because of its ability to chelate zinc and manganese. In the current study, we showed that neutrophils from calprotectin-deficient S100A9(-/-) mice have an impaired ability to inhibit Aspergillus fumigatus hyphal growth in vitro and in infected corneas in a murine model of fungal keratitis; however, the ability to inhibit hyphal growth was restored in S100A9(-/-) mice by injecting recombinant calprotectin. Furthermore, using recombinant calprotectin with mutations in either the Zn and Mn binding sites or the Mn binding site alone, we show that both zinc and manganese binding are necessary for calprotectin's antihyphal activity. In contrast to hyphae, we found no role for neutrophil calprotectin in uptake or killing of intracellular A. fumigatus conidia either in vitro or in a murine model of pulmonary aspergillosis. We also found that an A. fumigatus ∆zafA mutant, which demonstrates deficient zinc transport, exhibits impaired growth in infected corneas and following incubation with neutrophils or calprotectin in vitro as compared with wild-type. Collectively, these studies demonstrate a novel stage-specific susceptibility of A. fumigatus to zinc and manganese chelation by neutrophil-derived calprotectin.
authors.affiliation: Department of Ophthalmology and Visual Science, Case Western Reserve University, Cleveland, OH 44106; Department of Ophthalmology, University of California, Irvine, Irvine, CA 92697; Department of Physiology and Biophysics, University of California, Irvine, Irvine, CA 92697;
authors.forename: Heather L
authors.initials: HL
authors.lastname: Clark
authors.affiliation: Infectious Disease Service, Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10065;
authors.forename: Anupam
authors.initials: A
authors.lastname: Jhingran
authors.affiliation: Department of Ophthalmology and Visual Science, Case Western Reserve University, Cleveland, OH 44106;
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Ophthalmology and Visual Science, Case Western Reserve University, Cleveland, OH 44106;
authors.forename: Chairut
authors.initials: C
authors.lastname: Vareechon
authors.affiliation: Department of Ophthalmology and Visual Science, Case Western Reserve University, Cleveland, OH 44106;
authors.forename: Steven
authors.initials: S
authors.lastname: de Jesus Carrion
authors.affiliation: Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine, Nashville, TN 37232; Veterans Affairs Tennessee Valley Healthcare System, Nashville, TN 37212;
authors.forename: Eric P
authors.initials: EP
authors.lastname: Skaar
authors.affiliation: Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232; Center for Structural Biology, Vanderbilt University School of Medicine, Nashville, TN 37232; and.
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Chazin
authors.affiliation: Instituto de Biología Funcional y Genómica, Centro Mixto del Consejo Superior de Investigaciones Científicas y Universidad de Salamanca, 37002 Salamanca, Spain.
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Calera
authors.affiliation: Infectious Disease Service, Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10065;
authors.forename: Tobias M
authors.initials: TM
authors.lastname: Hohl
authors.affiliation: Department of Ophthalmology and Visual Science, Case Western Reserve University, Cleveland, OH 44106; Department of Ophthalmology, University of California, Irvine, Irvine, CA 92697; Department of Physiology and Biophysics, University of California, Irvine, Irvine, CA 92697; Eric.Pearlman@uci.edu.
authors.forename: Eric
authors.initials: E
authors.lastname: Pearlman
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D001232:Aspergillus fumigatus / Q000254:growth & development* / Q000276:immunology; D001692:Biological Transport / Q000235:genetics; D040501:Calgranulin A / Q000235:genetics / Q000378:metabolism; D040502:Calgranulin B / Q000235:genetics / Q000378:metabolism; D002614:Chelating Agents / Q000378:metabolism; D003315:Cornea / Q000276:immunology / Q000382:microbiology; D004195:Disease Models, Animal; D006801:Humans; D025301:Hyphae / Q000254:growth & development; D007634:Keratitis / Q000276:immunology / Q000382:microbiology*; D039841:Leukocyte L1 Antigen Complex / Q000235:genetics / Q000378:metabolism*; D008345:Manganese / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008875:Middle Aged; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism; D010587:Phagocytosis / Q000276:immunology; D055732:Pulmonary Aspergillosis / Q000276:immunology* / Q000382:microbiology; D013172:Spores, Fungal / Q000254:growth & development / Q000276:immunology; D055815:Young Adult; D015032:Zinc / Q000378:metabolism*
chemical_list: D040501:Calgranulin A; D040502:Calgranulin B; D002614:Chelating Agents; D039841:Leukocyte L1 Antigen Complex; C576329:S100A9 protein, mouse; C086145:S100a8 protein, mouse; D008345:Manganese; D015032:Zinc