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Topic:Neutrophils
Neutrophils are a type of white blood cell that play a significant role in the immune system of horses. They are involved in the body's defense mechanisms against infections and are typically one of the first responders to microbial invasion or tissue damage. Neutrophils function by engulfing and destroying pathogens through processes such as phagocytosis and the release of antimicrobial substances. In horses, the number and activity of neutrophils can be indicative of immune status and are often assessed in the context of infectious diseases, inflammation, and other health conditions. This page compiles peer-reviewed research studies and scholarly articles that explore the function, regulation, and clinical implications of neutrophils in equine health.
Equine neutrophil myeloperoxidase in plasma: design of a radio-immunoassay and first results in septic pathologies. The strangulated intestinal pathologies of horses are accompanied by a local activation of the neutrophils, that can be revealed by measuring the tissular enzymatic activity of the granulocytic enzyme myeloperoxidase (MPO). To estimate the possible spreading of this neutrophil activation to the systemic circulation, we designed a radioimmunoassay (RIA) for equine neutrophil myeloperoxidase (MPO) (EC 1.11.1.7) using a specific rabbit antiserum. MPO was labeled with 1 mCi 125I by a technique of self-labeling in the presence of 10(-4) M hydrogen peroxide. The RIA was performed by incubation of 10...
The role of neutrophil chemotactic cytokines in the pathogenesis of equine chronic obstructive pulmonary disease (COPD). Chronic obstructive pulmonary disease (COPD) is defined as a chronic obstructive inflammatory disease affecting the small airways associated with hay dust exposure (Lowell, F.C., 1964. Observation on heaves. An asthma like syndrome in the horse, J. Allergy 35, 322-330). The disease corresponds histopathologically to a chronic bronchiolitis (Gerber, H., 1973. Chronic pulmonary disease in the horse, Equine Vet. J. 5, 26-33; Winder, N.C., Grünig, G., Hermann, M., Howald, B., von Fellenberg, R., 1989. Comparison of respiratory secretion cytology and pulmonary histology in horses, J. Vet. Med., A3...
Flow cytometric determination of oxidative burst activity of equine peripheral blood and bronchoalveolar lavage-derived leucocytes. Flow cytometric techniques were developed for the evaluation of oxidative burst activity in equine peripheral blood neutrophils and lymphocytes, as well as bronchoalveolar lavage derived pulmonary alveolar macrophages and lymphocytes. The oxidation of dichlorofluorescin was measured by the increased fluorescence of cells stimulated with phorbol myristate acetate or a variety of other stimulants. Flow cytometry was a suitable method for the evaluation of the intracellular oxidation in all cell populations evaluated. Analysis was rapid and cell separation before analysis was not required. Hetero...
The flow cytometric evaluation of phagocytosis by equine peripheral blood neutrophils and pulmonary alveolar macrophages. Flow cytometry was used to assess the phagocytosis of fluorescent-labelled bacteria by equine peripheral blood neutrophils and pulmonary alveolar macrophages. Cell populations were prepared from venous blood following ammonium chloride lysis and from washed bronchoalveolar lavage derived samples. Discrete clusters of cells, corresponding to different leucocyte groups, were readily identified on the basis of differing light scattering properties and could thus be discriminated, negating the need for prior cell separation. Cells able to associate with fluorescent-labelled bacteria (by attachment...
Shear-induced platelet activation and platelet-neutrophil aggregate formation by equine platelets. To determine whether platelets become activated and form platelet-platelet or platelet-neutrophil aggregates, or both, when subjected to shear. Methods: Blood obtained from 3 Thoroughbreds. Methods: Blood, with PCV adjusted to 32 (low hematocrit) or 60 (high hematocrit)%, was subjected to shear rates of 11.25, 22.5, 45, 90, 225, and 750/s for 3 minutes by use of a cone-plate viscometer. Flow cytometric techniques were used to identify activated platelets, platelet-platelet aggregates, and platelet-neutrophil aggregates. Results: Shear resulted in decreased platelet count, increased mean platel...
Interactions between lipopolysaccharides and blood factors on the stimulation of equine polymorphonuclear neutrophils. In horses, the mechanisms of lipopolysaccharide (LPS) stimulation of isolated neutrophils to produce reactive oxygen species remain unknown. We re-investigated this problem by monitoring the luminol-enhanced chemiluminescence (CL) produced by LPS-stimulated equine neutrophils. The neutrophils were isolated from horse blood by discontinuous density gradient centrifugation (> or = 99% neutrophils; viability > or = 98%). Increasing concentrations of Escherichia coli (E. coli) LPS (from 0.01-10 microg ml(-1)) were used to activate the neutrophils. When LPS was used directly, without another ...
Inhibition of antigen-induced cutaneous responses of ponies with insect hypersensitivity by the histamine-1 receptor antagonist chlorpheniramine. A whole-body extract of Culicoides impunctatus induced a biphasic increase in oedema formation in ponies with insect hypersensitivity, with maxima after one and eight hours. The Culicoides antigen did not induce similar responses in ponies with no previous history of the disease. In insect-hypersensitive ponies the local administration of chlorpheniramine (12 micrograms) completely inhibited oedema formation in response to histamine (0.04 microgram) and to Culicoides antigen (0.5 microgram) at one hour, and the response to Culicoides antigen at eight hours was inhibited by 63 per cent. Chlorph...
Cytologic evaluation of bronchoalveolar lavage fluid from horses with recurrent airway obstruction after aerosol and parenteral administration of beclomethasone dipropionate and dexamethasone, respectively. To determine cytologic changes in horses with recurrent airway obstruction (heaves) after administration of aerosolized beclomethasone dipropionate and dexamethasone parenterally. Methods: 6 horses with inducible and reversible heaves. Methods: Episodes of heaves were induced by exposure to moldy hay and straw for 7 days. Horses were assigned to treatment groups (aerosolized beclomethasone, parenterally administered dexamethasone, aerosolized propellant), and pulmonary inflammation was evaluated by serial cytologic examination of bronchoalveolar lavage (BAL) fluid samples obtained on days 0, 7...
Inhaled leukotrienes cause bronchoconstriction and neutrophil accumulation in horses. Leukotrienes have been shown to mimic many of the pathophysiological processes in allergic airway disease. In this study the bronchoconstrictor effect of inhaled LTD4, and radiolabelled neutrophil accumulation in response to inhalation of LTB4, have been examined in the horse. In separate studies, solutions of LTD4 and LTB4 were administered to the airways of normal animals by nebulisation. LTD4, but not LTB4, caused a dose-dependent increase in pleural pressure which was maximal at three to four minutes and had returned to baseline by 15 to 20 minutes. On a molar basis LTD4 was 305 to 970 tim...
Effect of the 5-lipoxygenase inhibitor, fenleuton, on antigen-induced neutrophil accumulation and lung function changes in horses with chronic obstructive pulmonary disease. The leukotrienes (LT) LTD4 and LTB4 have been shown to cause bronchoconstriction and neutrophil accumulation, respectively, in horse lungs. Such changes are characteristic of the equine allergic respiratory disease, chronic obstructive pulmonary disease (COPD). To further investigate the role of these putative mediators in the pathogenesis of equine COPD the effect of a 5-lipoxygenase inhibitor, fenleuton, on antigen-induced changes in horses with this condition has been examined. Six horses with COPD underwent a series of four antigen challenges, one month apart, with placebo pre-treatment on...
Gelatinolytic activity in tracheal epithelial lining fluid and in blood from horses with chronic obstructive pulmonary disease. To determine whether gelatinolytic activity in tracheal epithelial lining fluid (TELF), blood neutrophils, and blood lymphocytes from horses was metalloprotease activity, and to compare, for healthy horses and horses with chronic obstructive pulmonary disease, gelatinolytic activity in neutrophils, lymphocytes, and serum with activity in TELF. Methods: 7 horses with chronic obstructive pulmonary disease (COPD) and 4 healthy control horses. Methods: Neutrophils and lymphocytes were obtained by means of Percoll separation. Zymography was used to detect gelatinolytic activity; EDTA inhibition and...
Equinins in equine neutrophils: quantification in tracheobronchial secretions as an aid in the diagnosis of chronic pulmonary disease. Equinins are a closely related group of proteins found in equine neutrophil granules. They demonstrate proteinase inhibiting activity restricted to microbial proteinase K and subtilisin, and they also possess antibacterial and antiviral properties. Antiproteinase K activity was measured in tracheobronchial secretions (TBS) of horses with mild (n = 15), moderate (n = 30) and severe (n = 16) chronic pulmonary disease, to determine its usefulness as an indicator of severity of disease and to measure neutrophil content. Determination of proteinase K inhibiting activity was based on a colorimetric ...
A dinucleotide mutation in the endothelin-B receptor gene is associated with lethal white foal syndrome (LWFS); a horse variant of Hirschsprung disease. Lethal white foal syndrome (LWFS) is a congenital anomaly of horses characterized by a white coat colour and aganglionosis of the bowel, which is similar to Hirschsprung disease (HSCR). We decided to investigate possible mutations of the endothelin-B receptor gene ( EDNRB ) in LWFS as recent studies in mutant rodents and some patients have demonstrated EDNRB defects. First, we identified a full-length cDNA for horse EDNRB . This cDNA fragment contained a 1329 bp open reading frame which encoded 443 amino acid residues. The predicted amino acid sequence was 89, 91 and 85% identical to human, bo...
Factors associated with survival in septicemic foals: 65 cases (1988-1995). In order to identify variables obtained at admission that could be used to predict survival in septicemic foals, medical records of 65 foals diagnosed with septicemia were reviewed. Initially, variables were analyzed independently (univariate analysis) for association with survival. Of the physical examination and historical data examined using univariate analysis, the ability to stand at admission, respiratory rate > or = 60 breaths per minute (bpm), and normal-appearing mucous membranes were significantly associated with survival. Foals with history of induced parturition were significant...
Evaluation of platelet activation and platelet-neutrophil aggregates in Thoroughbreds undergoing near-maximal treadmill exercise. To determine whether platelets become activated and form platelet-neutrophil aggregates during near-maximal treadmill exercise in horses. Methods: 4 Thoroughbreds. Methods: Horses were subjected to 4 standardized exercise tests on a treadmill, and blood samples were collected before exercise, at treadmill speed of 12 m/s, and 5 minutes after exercise. Flow cytometric techniques were used to identify activated platelets, and flow cytometric and microscopic techniques were used to identify platelet-neutrophil aggregates. Results: Platelet-neutrophil aggregates increased from 2.8 +/- 0.4% at rest...
Priming induces functional coupling of N-formyl-methionyl-leucyl-phenylalanine receptors in equine neutrophils. The synthetic formylpeptide fMLP is widely used as a model chemoattractant and secretagogue for mammalian neutrophils. Despite possessing fMLP receptors, equine neutrophils do not produce superoxide anions in response to fMLP and there is no inflammatory reaction in the horse when fMLP is injected intradermally. The functional capability of these receptors was investigated after pretreatment with recognized priming agents. Purified neutrophils were pretreated with lipopolysaccharide (LPS), platelet-activating factor (PAF), or tumor necrosis factor alpha (TNF-alpha) and superoxide anion generat...
Differential superoxide anion generation by equine eosinophils and neutrophils. Equine eosinophils and neutrophils are believed to play an important part in the protection of horses against parasitic and bacterial invasion. Eosinophils may also play a key role in the pathogenesis of equine inflammatory conditions such as the allergic skin disease, insect hypersensitivity. The factors which stimulate the respiratory burst of equine eosinophils and neutrophils are poorly understood. The first aim of the present study was to determine the effects of the phorbol ester, phorbol myristate acetate (PMA), which is believed to activate intracellular protein kinase C, and opsonised...
Evaluation of platelet activation and platelet-neutrophil aggregates in ponies with alimentary laminitis. To determine whether platelets are hyperaggregable or form platelet-neutrophil aggregates during the prodromal stages of acute laminitis of ponies. Methods: Healthy adult ponies: 8 experimental and 6 control. Methods: Acute laminitis was induced by oral administration of corn starch and wood flour to 8 ponies, and indices of platelet activation were evaluated. Blood samples were collected before and at 4, 8, 12, 24, 28, and 32 hours after carbohydrate administration, and PCV, total plasma protein concentration, platelet count, activated clotting time, whole blood recalcification time, spontane...
Influence of preoperative complete blood cell counts on surgical outcomes in healthy horses: 102 cases (1986-1996). To assess the value of CBC as a preoperative test in healthy horses undergoing cryptorchidectomy. Methods: Retrospective study. Methods: 117 horses. Methods: Medical records were reviewed to identify horses that had had cryptorchidectomy. Of the 117 horses identified, 102 were found that did not have a known medical condition believed to adversely affect surgery. Preoperative CBC was assessed in terms of abnormalities detected and effects of these abnormalities on development of complications during and after surgery and patient management. Results: Of 102 CBC performed, 55 contained abnormali...
Pharmacology of the 5-lipoxygenase inhibitors BAY Y 1015 and BAY X 1005 in the horse. Calcium ionophore A23187 induced time and concentration dependent production of immunoreactive leukotriene (LT) B4 by equine heparinized whole blood in vitro. Time dependent production of immunoreactive LTB4 by equine neutrophils and immunoreactive LTC4 by equine eosinophils in vitro was also demonstrated. The 5-lipoxygenase activating protein (FLAP) inhibitors, BAY X 1005 and BAY Y 1015, produced concentration dependent inhibition of ionophore-induced LTB4 synthesis by equine whole blood (mean +/- SEM IC50s n = 5; 6.14 +/- 0.28 microM vs. 12.30 +/- 0.75 microM for BAY Y 1015 and BAY X 1005, r...
Opsonic effect of equine plasma from different donors. The ability of equine plasma from different donors to enhance phagocytic capacity was assessed in neutrophils obtained from seven foals, aged 7-8 days (Study A), and from seven adult horses (Study B). Neutrophils were allowed to phagocytize fluorescent yeast cells opsonized with plasma from one of three donors or with pooled serum, all previously frozen (-18 degrees C) and thawed. The results were analysed by flow cytometry. In study A, fresh autologous foal serum was also used for opsonization, and in study B, heat-inactivated plasma and pooled serum were used in addition to untreated samples...
Effect of transportation on lower respiratory tract contamination and peripheral blood neutrophil function. To evaluate the effect of transportation on lower respiratory tract contamination and peripheral blood neutrophil function in horses and to compare results from transported horses with those obtained in earlier experiments from horses confined with heads elevated. Methods: A prospective study. Methods: Six horses were transported by road for 12 h. Clinical and haematological examination, transtracheal aspiration and cell function studies were conducted before and after transportation. Results obtained after transportation were compared to pre-transportation values. Results: After transportatio...
Functional characterization of equine neutrophils in response to calcium ionophore A23187 and phorbol myristate acetate ex vivo. Equine neutrophils (PMN) play a critical role in inflammatory processes in horses. The objective of this study was to characterize equine PMN function ex vivo following stimulation with calcium ionophore A23187 (A23187) and phorbol myristate acetate (PMA). These stimulants trigger different branches of the PMN activation process that occurs in vivo. Equine PMN were isolated from the whole blood of six clinically normal geldings using a one-step discontinuous Percoll gradient technique. Neutrophil aggregation, degranulation, and superoxide anion production were evaluated in assay systems which ...
Prognosis for neonatal foals in an intensive care unit. This study was conducted to develop an equation for the prediction of outcome in neonatal foals undergoing treatment in an intensive care unit (ICU). Fifty-three physical examination, historical, and clinicopathologic variables were analyzed from the records of 99 neonatal foals (< 14 days of age) treated in the neonatal ICU of the Equine Medical Center. The outcome was recorded and the results were categorized into either surviving or nonsurviving groups. The mean values for the 2 groups were compared, and variables that differed significantly between the two groups were retained and used ...
Effect of antigen challenge on the activation of peripheral blood neutrophils from horses with chronic obstructive pulmonary disease. The effect of antigen challenge on the state of activation of peripheral blood neutrophils from horses with chronic obstructive pulmonary disease (COPD) has been determined by measuring neutrophil superoxide anion formation. Prior to a seven-hour antigen challenge superoxide anion production by neutrophils from asymptomatic horses with COPD and normal horses in response to platelet activating factor (PAF) (with and without cytochalasin B), serum treated zymosan (STZ) and phorbol myristate acetate (PMA) was similar. Agonist-induced superoxide production by neutrophils from symptomatic COPD and ...
Gelatinolytic activity in tracheal aspirates of horses with chronic obstructive pulmonary disease. The gelatinolytic activity in tracheal aspirates (TA) of horses with chronic obstructive pulmonary disease (COPD) was analyzed using SDS-PAGE-gelatin-gel electrophoresis (zymography) and compared to TAs from healthy controls. The 110-90 kD MMP-9 type gelatinase was high in symptomatic disease phases (permanent disease 0.46 +/- 0.15, p < 0.001; or intermittent disease 0.47 +/- 0.12, p < 0.001) compared to healthy controls (0.10 +/- 0.07). Similarly, the overall gelatinolytic activity, the activity in high-mw gelatinolytic bands (210-190 and 150 kD) and in proteolytically processed fragmen...
Intrauterine fluid accumulation in oestrous mares. Intrauterine fluid (IUF) was collected using a tampon from mid-oestrous mares (n = 57) with and without ultrasonically detectable accumulations of free intraluminal fluid. Bacteria were cultured and neutrophils counted from all samples (n = 57). Total protein concentration, trypsin-inhibitor capacity (TIC), and plasmin, beta-glucuronidase (B-Gase) and N-acetyl-beta-D-glucosaminidase (NAGase) activities were determined in 27 IUF samples. The motility of spermatozoa in the presence of IUF, IUF extended with Kenney's medium (1:1) and Kenney's medium alone was analysed in 9 samples using a Hamilto...
Collagenolytic activity and its sensitivity to doxycycline inhibition in tracheal aspirates of horses with chronic obstructive pulmonary disease. The collagenolytic activity and its sensitivity to doxycycline inhibition in tracheal aspirates (TA) of horses with chronic obstructive pulmonary disease (COPD) was analyzed with SDS-PA gel electrophoresis (SDS-PAGE), using Type 1 collagen as the substrate. Both autoactive and total collagenase activities were significantly higher in TAs of horses with symptomatic COPD than in TAs of healthy horses. Doxycycline inhibition studies suggest that most of the TA collagenase is of the neutrophil type (MMP-8), but some is derived from other cells such as fibroblasts and monocyte/macrophages (MMP-1) a...
Effect of treatment with erythromycin on bronchoalveolar lavage fluid cell populations in foals. To determine whether oral administration of erythromycin alters the inflammatory response to bronchoalveolar lavage (BAL) in young horses. Methods: 12 healthy, unweaned, mixed-breed foals of either sex, between 2 and 4 months old. Methods: BAL was performed; 250 ml of phosphate-buffered saline solution (300 mOsm, pH 7.4) was administered in 50-ml aliquots. Foals were carefully monitored for 4 days, then erythromycin base (25 mg/kg of body weight, PO, q 12 h) was given to foals of the treated group. After 4 days, foals were reanesthetized, and the same lung was relavaged. Cytologic examination ...
Effect of activated equine neutrophils on sulfated proteoglycan metabolism in equine cartilage explant cultures. To determine the influence of activated equine neutrophils on sulfated glycosaminoglycan metabolism of equine articular cartilage in vitro. Methods: Articular cartilage explants harvested from the metacarpophalangeal joints of 7 horses. Methods: Proteoglycan degradation and synthesis were measured by release of glycosaminoglycan from the explants, and incorporation of [35S]sulfate into newly synthesized glycosaminoglycan. Results: Activated equine neutrophils significantly increased the release of glycosaminoglycan from explant matrix and the magnitude of that response was influenced by durati...
