title: Nonspecific cytotoxic cell antimicrobial protein (NCAMP-1): a novel alarmin ligand identified in zebrafish.
abstract: Cells from the coelomic cavity of adult zebrafish (zf) were used to study the alarmin-like activities of nonspecific cytotoxic cell antimicrobial protein-1 (NCAMP-1). Immunohistochemistry studies using polyclonal anti-NCAMP-1 identified constitutive NCAMP-1 in epithelial cells of the zf anterior kidney, in liver parenchyma and in the lamina propria of the intestine. NCAMP-1 was also located in the cytosol of mononuclear cells in these tissues. Cytosolic NCAMP-1 was detected in a diverse population of coelomic cells (CC) using confocal microscopy and polyclonal anti-NCAMP-1 staining. Large mononuclear and heterophil-like CC had intracellular NCAMP-1. These studies indicated that NCAMP-1 is constitutively found in epithelial cells and in ZFCC. To establish a relationship between NCAMP-1 and the alarmin functions of ATP, a stimulation-secretion model was initiated using zf coelomic cells (ZFCC). ZFCCs treated with the alarmin ATP secreted NCAMP-1 into culture supernatants. Treatment of ZFCC with either ATP or NCAMP-1 activated purinergic receptor induced pore formation detected by the ZFCC uptake of the dye YO-PRO-1. ATP induced YO-PRO-1 uptake was inhibited by antagonists oxidized-ATP, KN62, or CBB. These antagonists did not compete with NCAMP-1 induced YO-PRO-1 uptake. Binding of ZFCC by both ATP and NCAMP-1 produced an influx of Ca2+. Combined treatment of ZFCC with ATP and NCAMP-1 increased target cell cytotoxicity. Individually NCAMP-1 or ATP treatment did not produce target cell damage. Similar to ATP, NCAMP-1 activates cellular pore formation, calcium influx and cytotoxicity.
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
authors.forename: Margaret Mariscal
authors.initials: MM
authors.lastname: Monette
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
authors.forename: Donald Lee
authors.initials: DL
authors.lastname: Evans
authors.affiliation: Department of Veterinary Biosciences and Diagnostic Imaging, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
authors.forename: Thomas
authors.initials: T
authors.lastname: Krunkosky
authors.affiliation: Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
authors.forename: Alvin
authors.initials: A
authors.lastname: Camus
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
authors.forename: Liliana
authors.initials: L
authors.lastname: Jaso-Friedmann
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000067531:Alarmins / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D023181:Antimicrobial Cationic Peptides / Q000276:immunology / Q000378:metabolism* / Q000633:toxicity; D001583:Benzoxazoles / Q000737:chemistry / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005260:Female; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000473:pathology; D007668:Kidney / Q000378:metabolism / Q000473:pathology; D008024:Ligands; D008856:Microscopy, Fluorescence; D011806:Quinolinium Compounds / Q000737:chemistry / Q000378:metabolism; D058486:Receptors, Purinergic P2X7 / Q000737:chemistry / Q000378:metabolism; D015027:Zebrafish
chemical_list: D000067531:Alarmins; D000906:Antibodies; D023181:Antimicrobial Cationic Peptides; D001583:Benzoxazoles; D008024:Ligands; D011806:Quinolinium Compounds; D058486:Receptors, Purinergic P2X7; C089813:YO-PRO 1; D000255:Adenosine Triphosphate; D002118:Calcium