title: A halotyrosine antibody that detects increased protein modifications in asthma patients.
abstract: Airway inflammation has a pathophysiological role in asthma. Eosinophils, which are commonly increased in asthmatic airways, express eosinophil peroxidase and thereby produce hypobromite and bromotyrosine. Bromotyrosine is believed to be a specific marker for eosinophil activity, but developing an antibody against monobromotyrosine, the predominant brominated tyrosine residue found in vivo has proven difficult. We evaluated whether a 3-bromobenozoic acid hapten antigen produced antibodies that recognized halogenated tyrosine residues. Studies with small-molecule inhibitors or brominated or chlorinated protein suggested that a mouse monoclonal antibody (BTK-94C) selectively bound free and protein mono- and dibromotyrosine and, to a lesser degree, chlorotyrosine, and thus was designated a general halotyrosine antibody. We evaluated if this antibody had potential for characterizing human asthma using an enzyme-linked immunosorbent assay (ELISA) microarray platform to examine the halogenation of 23 proteins in three independent sets of sputum samples (52 samples total). In 15 healthy control or asthmatic subjects, ICAM, PDGF and RANTES had greater proportional amounts of halogenation in asthmatic subjects and the halogenation signal was associated with the severity of exercise-induced airway hyperresponsiveness. In 17 severe asthma patients treated with placebo or mepolizumab to suppress eosinophils, drug-related decreases in halogenation were observed with p values ranging from 0.006 to 0.11 for these 3 proteins. Analysis of 20 subjects that either had neutrophilic asthma or were healthy controls demonstrated a broad increase in halotyrosine (possibly chlorotyrosine) in neutrophilic asthmatics. Overall, these results suggest that an ELISA utilizing BTK-94C could prove useful for assessing airway inflammation in asthma patients.
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Hongjun
authors.initials: H
authors.lastname: Jin
authors.affiliation: Department of Medicine, University of Washington, Seattle, WA, USA.
authors.forename: Teal S
authors.initials: TS
authors.lastname: Hallstrand
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Don S
authors.initials: DS
authors.lastname: Daly
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Melissa M
authors.initials: MM
authors.lastname: Matzke
authors.affiliation: Department of Medicine, McMaster University, Hamilton, ON, Canada.
authors.forename: Parameswaran
authors.initials: P
authors.lastname: Nair
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Diana J
authors.initials: DJ
authors.lastname: Bigelow
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Joel G
authors.initials: JG
authors.lastname: Pounds
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA, USA. Electronic address: richard.zangar@pnnl.gov.
authors.forename: Richard C
authors.initials: RC
authors.lastname: Zangar
mesh_terms: D000293:Adolescent; D000328:Adult; D018927:Anti-Asthmatic Agents / Q000627:therapeutic use; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized / Q000627:therapeutic use; D001249:Asthma / Q000175:diagnosis* / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism / Q000503:physiopathology; D015415:Biomarkers / Q000378:metabolism; D016535:Bronchial Hyperreactivity; D016022:Case-Control Studies; D018946:Chemokine CCL5 / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004804:Eosinophils / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D054879:Halogenation; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism; D008875:Middle Aged; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D010982:Platelet-Derived Growth Factor / Q000276:immunology / Q000378:metabolism; D011237:Predictive Value of Tests; D011499:Protein Processing, Post-Translational; D016032:Randomized Controlled Trials as Topic; D012720:Severity of Illness Index; D013183:Sputum / Q000276:immunology / Q000378:metabolism; D016896:Treatment Outcome; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D055815:Young Adult
chemical_list: D018927:Anti-Asthmatic Agents; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D015415:Biomarkers; C483024:CCL5 protein, human; D018946:Chemokine CCL5; C573420:ICAM1 protein, human; D010982:Platelet-Derived Growth Factor; C475724:bromotyrosine; D018799:Intercellular Adhesion Molecule-1; D014443:Tyrosine; C434107:mepolizumab
keywords: Asthma; Bromotyrosine; EIB; Eosinophil; Halotyrosine; Neutrophil