Antigen-mediated pulmonary eosinophilia in immunoglobulin G1-sensitized guinea pigs: eosinophil peroxidase as a simple specific marker for detecting eosinophils in bronchoalveolar lavage fluid.
Eosinophil peroxidase (EPO) has been used previously to detect the number of eosinophils in the peritoneal exudate and bone marrow of mice. The present study was undertaken to determine 1) whether EPO activity may provide a measure of a change in eosinophils in bronchoalveolar lavage fluid (BALF) of guinea pigs, 2) whether immunoglobulin (Ig)G1 could play a role in pulmonary eosinophilia and 3) effects of pharmacological agents on the EPO response in an IgG1 passively sensitized animal model. The activity of EPO was assessed by the ability of cell lysates (0.1% Triton-100 treatment) to oxidize 1 mM o-phenylenediamine in the presence of 1 mM H2O2 for 5 min at 22 degrees C. The enzyme activity was found to be eosinophil dependent, inhibited by the EPO inhibitor 3-amino-1,2,4-triazole (IC50 = approximately 0.1 mM) and relatively resistant to heat treatment (no loss of activity after 2-hr preincubation at 56 degrees C). To determine antigen-dependent eosinophil and EPO responses, guinea pigs were passively sensitized i.p. with 0.5 mg/kg of an affinity-purified antiovalbumin (OA) IgG1. Two to 3 days later, the sensitized animals were injected with pyrilamine (5 mg/kg, i.p.) before OA aerosol challenge. Aerosolized OA (0.1%) caused a significant increase in both eosinophil number and EPO activity in BALF of sensitized guinea pigs at 18 to 24 hr post-challenge. At a given concentration of aerosolized OA, the enzyme activity increased as a function of the antibody dose and time post-OA challenge.(ABSTRACT TRUNCATED AT 250 WORDS)
Cheng
J B
JB
Department of Immunology and Infectious Diseases, Pfizer Inc, Groton, Connecticut.
Pillar
J S
JS
Shirley
J T
JT
Showell
H J
HJ
Watson
J W
JW
Cohan
V L
VL
D000818:Animals; D000941:Antigens / Q000276:immunology*; D015415:Biomarkers / Q000032:analysis*; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology* / Q000201:enzymology; D047209:Eosinophil Peroxidase; D004804:Eosinophils / Q000201:enzymology*; D005260:Female; D006168:Guinea Pigs; D007074:Immunoglobulin G / Q000276:immunology*; D007962:Leukocytes / Q000201:enzymology; D010047:Ovalbumin / Q000276:immunology; D010544:Peroxidases / Q000032:analysis*; D010972:Platelet Activating Factor / Q000494:pharmacology; D011657:Pulmonary Eosinophilia / Q000175:diagnosis / Q000209:etiology*
D000941:Antigens; D015415:Biomarkers; D007074:Immunoglobulin G; D010972:Platelet Activating Factor; D010047:Ovalbumin; D047209:Eosinophil Peroxidase; D010544:Peroxidases