[en] The aim of this work was to measure the myeloperoxidase (MPO) concentration in bronchoalveolar lavage (BAL) fluid collected from horses with recurrent airway obstruction (RAO), both in crisis and in remission, as well as from healthy horses. Seven horses with RAO were exposed to moldy hay until the maximum change in pleural pressure was greater than 1.5 kPa. At that point, BAL was performed, and the total cell counts and percentages in the fluid were immediately determined. To measure the MPO concentration in BAL-fluid supernatant, we used a specific enzyme-linked immunosorbent assay with polyclonal antibodies against equine MPO. The tests were repeated on the horses with RAO after they had spent 2 mo on pasture. Six healthy horses serving as controls underwent the same tests. The absolute and relative neutrophil counts and the MPO concentration in the BAL fluid were significantly greater in the horses with an RAO crisis than in the control horses. After 2 mo on pasture, the horses that had been in RAO crisis were clinically normal, and their neutrophil counts and MPO levels in BAL fluid had significantly decreased; during remission their neutrophil counts were not significantly different from those in the healthy horses, but their MPO concentration remained significantly higher. This study showed that determining the MPO concentration in a horse's BAL fluid is technically possible and that during remission from RAO the concentration remains higher than normal. Thus, MPO may be a marker of neutrophil presence and activation in the lower airways. [fr] Le but de cette étude était de mesurer la concentration de myéloperoxydase (MPO) dans le liquide de lavage broncho-alvéolaire (BAL) récolté chez des chevaux avec obstruction respiratoire récurrente (RAO), en période de crise et en rémission, ainsi que chez des chevaux en santé. Sept chevaux avec RAO ont été exposés à du foin moisi jusqu’à ce que le changement maximal dans la pression pleurale soit plus grand que 1,5 kPa. À cet instant on procéda au BAL et le dénombrement cellulaire total de même que les pourcentages dans le liquide furent immédiatement déterminés. Afin de mesurer la concentration de MPO dans le liquide de BAL on procéda à une épreuve immunoenzymatique spécifique utilisant des anticorps polyclonaux dirigés contre MPO. Les épreuves ont été répétées chez les chevaux avec RAO après qu’ils eurent passé 2 mois au pâturage. Six chevaux en santé servant de témoin ont subi les mêmes tests. Les comptes absolus et relatifs de neutrophiles et les concentrations de MPO dans le liquide de BAL étaient significativement plus élevés chez les chevaux avec RAO en période de crise que chez les chevaux témoins. Après 2 mois au pâturage, les chevaux qui avaient été en crise de RAO étaient cliniquement normaux et leurs comptes en neutrophiles et niveaux de MPO dans le liquide de BAL avaient diminué de manière significative; durant la rémission les comptes de neutrophiles n’étaient pas significativement différents de ceux des chevaux témoins, mais les concentrations de MPO sont demeurées significativement plus élevées. Cette étude a permis de démontrer qu’il était techniquement possible de déterminer la concentration de MPO dans du liquide de BAL équin et que durant la période de rémission d’une crise de RAO la concentration est demeurée plus élevée que la normale. Ainsi, la MPO pourrait être un marqueur de la présence et de l’activation des neutrophiles dans les voies respiratoires inférieures.
(Traduit par Docteur Serge Messier)
Disciplines :
Veterinary medicine & animal health Anatomy (cytology, histology, embryology...) & physiology
Author, co-author :
Art, Tatiana ; Université de Liège - ULiège > Département de sciences fonctionnelles > Phys. neuro-muscul., de l'effort - Méd. sport. des animaux
Franck, Thierry ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Lekeux, Pierre ; Université de Liège - ULiège > Département de sciences fonctionnelles > Physiologie
De Moffarts, Brieuc
Couëtil, Laurent
Becker, Martine
Kohnen, Serge
Deby-Dupont, Ginette
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 :
Myeloperoxidase concentration in bronchoalveolar lavage fluid from healthy horses and those with recurrent airway obstruction
Klebanoff SJ. Myeloperoxidase: friend and foe. J Leukoc Biol 2005;77:598-625.
Serteyn D, Grulke S, Franck T, Mouithys-Mickalad A, Deby-Dupont G. La myéloperoxydase des neutrophiles, une enzyme de défense aux capacités oxydantes. Ann Méd Vét 2003;147:79-93.
Schmekel B, Karlsson SE, Linden M, Sundtrom C, Tegner H, Venge P. Myeloperoxidase in human lung lavage. I. A marker of local neutrophil activity. Inflammation 1990;14:447-454.
Thukkani AK, Martinson BD, Albert CJ, Vogler GA, Ford DA. Neutrophil-mediated accumulation of 2-CIHDA during myocardial infarction: 2-CIHDA-mediated myocardial injury. Am J Physiol Heart Circ Physiol 2005;288:H2955-H2964.
Deby-Dupont G, Grulke S, Caudron I, et al. Equine neutrophil myeloperoxidase in plasma: design of a radio-immunoassay and first results in septic pathologies. Vet Immunol Immunopathol 1998;66:257-271.
Mathy-Hartert M, Bourgeois E, Grulke S, et al. Purification of myeloperoxidase from polymorphonuclear leucocytes. Can J Vet Res 1998;62:127-132.
Yarbrough B, Snyder JR, Harmon FA, O'Connell KA. Evaluation of myeloperoxidase concentrations in experimentally induced equine colonic ischaemia and reperfusion. Equine Vet J 1994; 26:67-69.
McConnico RS, Weinstock D, Poston ME, Roberts MC. Myeloperoxidase activity of the large intestine in an equine model of acute colitis. Am J Vet Res 1999;60:807-813.
Grulke S, Benbarek H, Caudron I, et al. Plasma myeloperoxidase level and polymorphonuclear leukocyte activation in horses suffering from large intestinal obstruction requiring surgery: preliminary results. Can J Vet Res 1999;63:142-147.
McGorum BC, Dixon PM, Halliwell REW. Response of horses affected with chronic obstructive pulmonary disease to inhalation challenges with mould antigens. Equine Vet J 1993;25: 261-267.
Robinson NE. International Workshop on Equine Chronic Airway Disease. Michigan State University, 16-18 June 2000. Equine Vet J 2001;33:5-19.
Franck T, Grulke S, Deby-Dupont G, et al. Development of an enzyme-linked immunosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood. J Vet Diagn Invest 2005;17:412-419.
Krawisz JE, Sharon P, Stenson WF. Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models. Gastroenterology 1984;87:1344-1350.
Weiland JE, Davis WB, Holter JF, Mohammed JR, Dorinsky PM, Gadek JE. Lung neutrophils in the adult respiratory distress syndrome. Clinical and pathophysiologic significance. Am Rev Respir Dis 1986;133:218-225.
Lamb NJ, Gutteridge JM, Baker C, Evans TW, Quinlan GJ. Oxidative damage to proteins of bronchoalveolar lavage fluid in patients with acute respiratory distress syndrome: evidence for neutrophil-mediated hydroxylation, nitration, and chlorination. Crit Care Med 1999;27:1738-1744.
Brazil TJ. The role of neutrophils in equine heaves. In: Proceedings of the 2nd World Equine Airways Symposium and 19th Comparative Respiratory Society Meeting; 2001 July 19-23; Edinburgh, Scotland. Guelph, Ontario: Lifelearn, 2001 [CD-ROM].
Léguillette R. Recurrent airway obstruction - heaves. Vet Clin Equine 2003;19:63-86.
Olszewski M, Laber G. Production of free oxygen radicals by phagocytes from respiratory tract lavaged as well as from peripheral blood of horses with chronic obstructive pulmonary disease (COPD) in comparison to healthy animals. Wien Tierarztl Monatsschr 1993;80:332-337.
Koivunen AL, Maisi P, Konttinen YT, Prikk K, Sandholm M. Collagenolytic activity and its sensitivity to doxycycline inhibition in tracheal aspirates of horses with chronic obstructive pulmonary disease. Acta Vet Scand 1997;38:9-16.
Raulo SM, Sorsa TA, Kiili MT, Maisi PS. Evaluation of collagenase activity, matrix metalloproteinase-8, and matrix metalloproteinase-13 in horses with chronic obstructive pulmonary disease. Am J Vet Res 2001;62:1142-1148.
Art T, Kirschvink N, Smith N, Lekeux P. Indices of oxidative stress in blood and pulmonary epithelium lining fluid in horses suffering from recurrent airway obstruction. Equine Vet J 1999;31:397-401.
Olszewski MA, Robinson NE, Zhu FX, Zhang XY, Tithof PK. Mediators of anaphylaxis but not activated neutrophils augment cholinergic responses of equine small airways. Am J Physiol 1999;276:L522-L529.
Robinson NE. Recurrent airway obstruction (heaves). In: Lekeux P, ed. Equine Respiratory Diseases [document B0317.1101]. Ithaca, New York: International Veterinary Information Service, 2001; available at www.ivis.org/special_books/Lekeux/robinson/chapter_frm.asp?LA=1 (accessed 2006 July 4).
Marr KA, Foster AP, Lees P, Cunningham FM, Page CP. Effect of antigen challenge on the activation of peripheral blood neutrophils from horses with chronic obstructive pulmonary disease. Res Vet Sci 1997;62:253-260.
Marr KA, Lees P, Cunningham FM. Antigen challenge increases adherence of circulating neutrophils in horses with chronic obstructive pulmonary disease. Equine Vet J 2002;34:65-70.
Rickards KJ, Page CP, Lees P, Cunningham FM. Phosphodiesterase activity in neutrophils from horses with chronic obstructive pulmonary disease. Vet Immunol Immunopathol 2000;76:319-330.