[en] Equine inflammatory disease is accompanied by a neutrophil activation resulting in the release of granulocytic enzyme myeloperoxidase (MPO). To measure MPO in horse plasma as marker of neutrophil activation, the authors purified equine neutrophil MPO and developed a specific enzyme immunoassay using 2 specific polyclonal antibodies obtained from rabbit (primary antibody) and guinea pig (secondary antibody). The sandwich complex "primary antibody-MPO-secondary antibody" was detected using a goat anti-guinea pig immunoglobulin antibody conjugated to alkaline phosphatase. The enzyme-linked immunosorbent assay (ELISA) showed good precision and accuracy, with intra- and interassay coefficients of variation below 10% for MPO concentrations ranging from 0.78 to 50 ng/ml. A stable plasma MPO value, unaffected by time elapsed between blood collection and centrifugation, was obtained with plasma from EDTA anticoagulated blood. The mean MPO value measured in 38 healthy horses was 181.80 +/- 64.74 ng/ml. In 20 horses suffering from obstruction of the large or small intestine, MPO concentrations measured at the time of arrival at the intensive care unit were significantly higher than mean normal value, ranging from 477.88 to 2,748.13 ng/ml. Work is in progress to apply this MPO ELISA technique to other biological fluids and other equine diseases.
Research Center/Unit :
CORD - Centre de l'Oxygène, Recherche et Développement - ULiège
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Franck, Thierry ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Grulke, Sigrid ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Département clinique des animaux de compagnie et des équidés
Deby-Dupont, Ginette; Université de Liège - ULiège > Centre de l'Oxygène : Recherche et Développement (C.O.R.D.)
Deby, Carol; Centre de Biochimie de l'Oxygène
Duvivier, Hania
Péters, Fabrice
Serteyn, Didier ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Language :
English
Title :
Development of an enzyme-linked immunosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood
Publication date :
September 2005
Journal title :
Journal of Veterinary Diagnostic Investigation
ISSN :
1040-6387
Publisher :
Amer Assoc Veterinary Laboratory Diagnosticians Inc, Turlock, United States - California
Volume :
17
Issue :
5
Pages :
412-419
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
MPOEQUI
Funders :
DGTRE - Région wallonne. Direction générale des Technologies, de la Recherche et de l'Énergie
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Bibliography
Andrews PC, Krinsky NI: 1980, The reductive cleavage of myeloperoxidase in half, producing enzymatically active hemi-myeloperoxidase. J Biol Chem 256:4211-4218.
Benbarek H, Deby-Dupont G, Caudron I, et al.: 1998, Interaction between lipopolysaccharides and blood factors on the stimulation of equine polymorphonuclear neutrophils. Vet Immunol Immunopathol 64:313-322.
Borregaard N, Cowland JB: 1997, Granules of the human neutrophilic polymorphonuclear leukocytes. Blood 89:3503-3521.
Brennan ML, Hazen SL: 2003, Emerging role of myeloperoxidase and oxidant stress markers in cardiovascular risk assessment. Curr Opin Lipidol 14:353-359.
Carr AC, Myzak MC, Stocker R, et al.: 2000, Myeloperoxidase binds to low density lipoprotein: potential implications for atherosclerosis. FEBS Lett 487:176-180.
Cooray R, Peterson CGB, Holmberg O: 1993, Isolation and purification of bovine myeloperoxidase from neutrophil granules. Vet Immunol Immunopathol 38:261-272.
Cooray R, Petersson CGB, Moberg L, Bage R: 1995, A sensitive double-antibody enzyme-linked immunosorbent assay for bovine myeloperoxidase and its application to serum and neutrophil extracts. J Vet Med 42:481-491.
Dagleish MR, Brazil TJ, Scudamore CL: 2003, Potentiation of the extracellular release of equine neutrophil elastase and alpha-1-proteinase inhibitor by a combination of two bacterial cell wall components: fMLP and LPS. Equine Vet J 35:35-39.
Deaton CM, Marlin DJ, Smith NC, et al.: 2004, Breath condensate hydrogen peroxide correlates with both airway cytology and epithelial lining fluid ascorbic acid concentration in the horse. Free Radic Res 38:201-208.
Deby-Dupont G, Deby C, Lamy M: 1999, Neutrophil myeloperoxidase revisited: it's role in health and disease. Intensivmed 36:500-513.
Deby-Dupont G, Grulke S, Caudron I, et al.: 1998, Equine myeloperoxidase in plasma: design of a radio-immunoassay and first results in septic pathologies. Vet Immunol Immunopathol 66:257-271.
Doerner KC, White BA: 1990, Detection of glycoproteins separated by nondenaturing polyacrylamide gel electrophoresis using the periodic acid-Schiff Stain. Anal Biochem 187:147-150.
Graff G, Gamache DA, Brady MT, et al.: 1998, Improved myeloperoxidase assay for quantitation of neutrophil influx in a rat model of endotoxin-induced uveitis. J Pharmacol Toxicol Methods 39:169-178.
Griffin SV, Chapman PT, Lianos EA, Lockwood CM: 1999, The inhibition of myeloperoxidase by ceruloplasmin can be reversed by anti-myeloperoxidase antibodies. Kidney Int 55:917-925.
Grulke S, Benbarek H, Caudron I, et al.: 1999, Plasma myeloperoxidase level and polymorphonuclear leukocyte activation in horses suffering from large intestinal obstruction requiring surgery: preliminary results. Can J Vet Res 63:142-147.
Haqqani AS, Sandhu JK, Birnboim HC: 1999, A myeloperoxidase-specific assay based upon bromide-dependent chemiluminescence of luminol. Anal Biochem 273:126-132.
Harrison JE, Pabalan S, Schultz J: 1977, The subunit structure of crystalline canine myeloperoxidase. Biochem Biophys Acta 493:247-259.
Hazen SL, Hsu FF, Duffin K, Heinecke JW: 1996, Molecular chlorine generated by the myeloperoxidase-hydrogen peroxide-chloride system of phagocytes converts low density lipoprotein cholesterol into a family of chlorinated sterols. J Biol Chem 271:23080-23088.
Iwamoto H, Morita Y, Kobayashi T, Hasegawa E: 1988, Subunit structures of three human myeloperoxidases. J Biochem 103:688-692.
Kaminski KA, Bonda TA, Korecki J, Musial WJ: 2002, Oxidative stress and neutrophil activation - the two keystones of ischemia/reperfusion injury. Int J Cardiol 86:41-59.
Klebanoff SJ: 1991, Myeloperoxidase. Proc Assoc Am Physicians 111:383-389.
Mathy-Hartert M, Bourgeois I, Grulke S, et al.: 1998, Purification of myeloperoxidase from equine polymorphonuclear leucocytes. Can J Vet Res 62:127-132.
Mathy-Hartert M, Deby-Dupont G, Deby C, et al.: 1995, Cytotoxicity towards human endothelial cells, induced by neutrophil myeloperoxidase: protection by ceftazidime. Mediators Inflamm 4:437-443.
Mathy-Hartert M, Deby-Dupont G, Melin P, et al.: 1996, Cultured macrophages acquire a bactericidal activity against Pseudomonas aeruginosa after incorporation of myeloperoxidase. Experientia 52:167-174.
Maxey KM, Maddipati KR, Birkmeier J: 1992, Interference in enzyme immunoassay. J Clin Immunoassay 15:116-120.
McConnico RS, Weinstock D, Poston ME, Roberts MC: 1999, Myeloperoxidase activity of the large intestine in an equine model of acute colitis. Am J Vet Res 60:807-813.
Monzani E, Bonafe B, Fallarini A, et al.: 2001, Enzymatic properties of human hemalbumin. Biochim Biophys Acta 1547:302-312.
Moore RM, Muir WW, Granger DN: 1995, Mechanisms of gastrointestinal ischemia-reperfusion injury and potential therapeutic interventions: a review and its implication in the horse. J Vet Intern Med 9:115-132.
Olsen RL, Little C: 1984, Studies on the subunits of human myeloperoxidase. Biochem J 222:701-709.
Olsen RL, Steigen TK, Holm T, Little C: 1986, Molecular forms of myeloperoxidase in human plasma. Biochem J 237:559-565.
Omi H, Okayama N, Shimizu M, et al.: 2002, Participation of high glucose concentrations in neutrophil adhesion and surface expression of adhesion molecules on cultured human endothelial cells: effect of antidiabetic medicines. J Diabetes Complications 16:201-208.
Pember SO, Fuhrer-Krusi SM, Barnes KC, Kinkade JM: 1982, Isolation of three native forms of myeloperoxidase from human polymorphonuclear leukocytes. FEBS Lett 140:103-108.
Pincemail J, Deby-Dupont G, Deby C, et al.: 1991, Fast double antibody radioimmunoassay of human granulocyte myeloperoxidase and its application to plasma. J Immunol Methods 137:181-191.
Segelmark M, Persson B, Hellmark T, Wieslander J: 1997, Binding and inhibition of myeloperoxidase (MPO): a major function of ceruloplasmin. Clin Exp Immunol 108:167-174.
Taylor KL, Guzman GS, Pohl J, Kinkade JM: 1990, Distinct chromatographic forms of human hemi-myeloperoxidase obtained by reductive cleavage of the dimeric enzyme. J Biol Chem 265:1538-1546.
Weiss DJ, Evanson OA, McClenahan D, et al.: 1997, Evaluation of platelet activation and platelet-neutrophil aggregates in ponies with alimentary laminitis. Am J Vet Res 58:1376-1380.
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