Leukocytes, also known as white blood cells, are an integral component of the equine immune system, responsible for defending horses against infections and foreign substances. These cells are produced in the bone marrow and circulate throughout the bloodstream and tissues. In horses, leukocytes are categorized into several types, including neutrophils, lymphocytes, monocytes, eosinophils, and basophils, each with specific functions in immune response. Neutrophils are involved in phagocytosis and the destruction of pathogens, while lymphocytes are key players in adaptive immunity. Monocytes differentiate into macrophages and dendritic cells, contributing to pathogen clearance and antigen presentation. Eosinophils and basophils are associated with allergic reactions and parasitic infections. Variations in leukocyte counts and distribution can indicate underlying health conditions, making them valuable in veterinary diagnostics. This page aggregates peer-reviewed research studies and scholarly articles that explore the physiology, function, and diagnostic relevance of leukocytes in equine health.
Kotilainen T, Huhtinen M, Katila T.The objective of this study was to investigate the inflammatory reaction induced in the equine uterus by insemination with fresh and frozen semen. Eleven groups (6 to 8 mares per group) were studied during 2 breeding seasons. The mares were inseminated using raw semen, frozen semen, extended fresh and frozen semen, concentrated fresh semen, seminal plasma and seminal extenders only. One group was bred naturally. Six hours after insemination, the uteri were flushed with 50 ml of phosphate-buffered saline (PBS). Seventeen out of 104 samples (16%) exhibited slight bacterial growth. Neutrophil con...
Waelchli RO, Lutz H, Hermann M, Rüegg C, Eggenberger E.Hematologic reference values were established in 18 healthy foals in the first two months of life. Blood samples were collected prior to colostrum consumption and at 30 hours, 1 week, 3, 5, 7 and 9 weeks of age. PCV, Hb and RBC decreased during the first week and RBC, but not PCV and Hb, increased toward the end of the second month. With the exception of the sample at 1 week, the foals had mean RBC values significantly higher than those of controls. Mean MCV and MCH did not change during the first week, but decreased slightly thereafter; all means were smaller than in controls. The numbers of ...
Lester GD, Alleman AR, Raskin RE, Meyer JC.Pancytopenia was observed in two 3-year-old geldings and one 11-year-old mare. All horses had a brief history (2 days to 4 weeks) of fever, anorexia, and depression. One of the three horses had blast cells present on a peripheral blood smear. Examination of the bone marrow showed substantial infiltration with neoplastic lymphoid cells. At necropsy, neoplastic cells were restricted to the bone marrow in one horse, present in bone marrow, liver, and spleen in the second horse, and reported in multiple tissues in the third horse, including bone marrow, kidneys, lung, myocardium and lymph nodes. T...
McCulloch J, Williamson SA, Powis SJ, Edington N.It has been suggested that EHV-1 infection may perturb immune responsiveness in the natural equine host. The mechanism underlying this phenomenon is not clear, but disturbances of circulating leucocyte populations could contribute. In order to objectively assess the nature of the haematological changes provoked by EHV-1 infection, two groups of conventionally-maintained Welsh mountain ponies were challenge-infected intra-nasally with the Ab4 isolate of EHV-1. These groups were controlled by similarly-sized groups of non-infected ponies. All data generated was subjected to rigorous statistical ...
Fairbairn SM, Page CP, Lees P, Cunningham FM.Previous studies have shown that bronchoalveolar lavage fluid from horses with allergic respiratory disease and showing clinical symptoms contains increased numbers of neutrophils. In some cases, the eosinophil count is also increased. In this study the time course of changes in lung function and the accumulation of radiolabelled leucocytes and platelets in the lungs of allergic and normal horses has been examined during a 7 hr allergen exposure. Antigen challenge had no effect on pleural pressure or the distribution of radiolabelled neutrophils, eosinophils or platelets in normal horses. In c...
Sullivan KE, Snyder JR, Madigan JE, Pascoe JR, Farver TB, Thurmond MC, Andresen JW.Granulocyte colony-stimulating factor (G-CSF) is a glycoprotein that regulates the proliferation and maturation of hematopoietic progenitor cells and modulates the function of mature neutrophils. The responses to administration of G-CSF alone, and in combination with antimicrobials, were studied in an equine model of ascending colon ischemia. Complete segmental colonic ischemia (3.75 hours) with pelvic flexure enterotomy was created in four treatment groups. Group 1 horses received recombinant canine G-CSF (10 micrograms/kg, every 24 hours, intramuscularly), gentamicin sulfate (2.2 mg/kg, ever...
Messick JB, Rikihisa Y.The binding, internalization, and proliferation of Ehrlichia risticii in P388D1 cells and equine polymorphonuclear (PMN) leukocytes were studied by immunofluorescent staining and flow cytometric analysis. The binding of ehrlichiae to P388D1 cells at 4 degrees C was dose dependent, and the antigens of bound organisms were susceptible to pronase treatment. Additionally, the binding of ehrlichiae to P388D1 cells was diminished when either P388D1 cells or ehrlichiae were treated with 1% paraformaldehyde for 30 min or 0.25% trypsin for 15 min. These results indicate that the ehrlichial ligand and h...
Pemberton AD, Miller HR, John HA, Scudamore CL.1. Antiproteinase deficiency can result in excessive proteinase-induced tissue damage. The major anti-elastase (Spi1) protein of equine alpha 1-proteinase inhibitor (alpha 1-PI) was isolated from the plasma/serum of three common haplotypes (I, L and U). 2. The N-terminal amino acid sequences of the three inhibitors were identical, but were only approx 65-77% homologous with two other published equine Spi1 sequences. 3. All three inhibitors complexed quickly and irreversibly with equine leucocyte proteinase 2A (kass = 2 x 10(7) M-1 sec-1). They were also efficient inhibitors of chymase (rat mas...
Madigan JE.Equine ehrlichiosis is a seasonal disease of horses first reported in 1969. Clinical signs in horses include high fever, depression, partial hypophagia, anorexia, limb edema, petechiation, icterus, ataxia, and reluctance to move. Hematologic changes include leukopenia, thrombocytopenia, icterus, anemia, and inclusion bodies, principally in neutrophils and occasionally in eosinophils. Diagnosis is made by clinical signs and observing characteristic morulae in a blood smear with standard Wright's stain. Mortality is low unless secondary infection develops or injury occurs as a result of incoordi...
Stockert JC, Trigoso CI, Tato A, Ferrer JM.The structure of specific granules from horse eosinophil leukocytes is still largely unknown. In this work, electron microscopical studies of horse eosinophils reveal that the large cytoplasmic granules contain an external membrane, a matrix of less density, and a dense (non crystalline) core. Round vacuolar inclusions of matrix materials were often observed within the cores. Horse eosinophil granules showed a considerable heterogeneity, and three morphological types could be identified according to structural features of the core and matrix.
Kordula T, Dubin A, Schooltink H, Koj A, Heinrich PC, Rose-John S.Horse blood leucocytes contain an elastase inhibitor (HLEI) belonging to the serpin family. Poly(A)+RNA isolated from these cells was used to construct a cDNA library in lambda gt10, which was first screened with a synthetic degenerate oligonucleotide probe corresponding to the amino acid sequence of the reactive centre of the inhibitor. Three clones were obtained covering the entire coding region of the protein. Sequencing of these clones showed identity with the amino acid sequence obtained from Edman degradation of the elastase inhibitor. The coding sequence of the HLEI cDNA was cloned into...
Sellon DC, Cullen JM, Whetter LE, Gebhard DH, Coggins L, Fuller FJ.An IgG1 mouse monoclonal antibody, designated 1.646, is described which recognizes a cytoplasmic antigen of equine mononuclear phagocytes. Indirect fluorescent antibody staining of peripheral blood leukocytes reveals a granular cytoplasmic staining, predominantly in adherent blood mononuclear cells. Indirect fluorescent antibody staining is positive for alveolar and peritoneal macrophages. In some horses, a few neutrophils are also stained. In equine tissue samples stained by immunohistochemistry, the distribution of positive cells is consistent with the distribution of tissue macrophages. The...
Ainsworth DM, Weldon AD, Beck KA, Rowland PH.Five 3-month-old foals presenting with fever and respiratory disease were found to have pulmonary abscesses with patchy to diffuse alveolar and interstitial pneumonia on post-mortem examination. All affected foals had evidence of Rhodococcus equi infection and had few to abundant Pneumocystis carinii cysts in the sections of affected lung. Of the 5 foals examined radiographically, 3 had a distinct reticulonodular (miliary) pattern which may aid in the ante-mortem diagnosis of P. carinii pneumonia (PCP). Leukocyte counts of foals with PCP were significantly greater than in the control group of ...
Wilson DV, Eberhart SW, Robinson NE, Rice R, Gray PR.The effects of exogenous platelet-activating factor (PAF) were determined in anesthetized ponies. Administration of PAF induced a decrease in cardiac index that resulted in systemic hypotension. This was followed by tachycardia, hypertension, and a return of cardiac index to baseline. Pulmonary arterial pressure increased markedly because of pulmonary vasoconstriction. Exogenous PAF also caused leukopenia and thrombocytopenia. The specific PAF receptor antagonist (WEB 2086) blocked all PAF-induced changes. Flunixin meglumine, a cyclooxygenase inhibitor, abolished the pulmonary hypertension and...
Lunn DP.Monoclonal antibody technology has allowed the recognition and study of numerous leucocyte antigens in man and laboratory animals for over a decade. Numerous advances in the understanding of immune responses and immunopathology have resulted. In recent years equine researchers have started to develop similar reagents, which now offer a powerful tool to investigators of equine immunology and disease.
Kramer J, Tornquist S, Erfle J, Sloeojan G.Large granular lymphocytes (LGL) occur as a leukemic or aleukemic neoplasia in humans,(1) rats,(2) cats,(3,4) dogs,(5) and horses.(6) The single case of equine LGL previously reported was aleukemic.(5) In contrast to this previous report of aleukemic equine LGL neoplasia, the leukemia in the case presented here established the antemortem diagnosis of LGL.
Lunn DP, Holmes MA, Duffus WP.This paper describes the characteristics of a monoclonal antibody (CVS10) that reacts with an equine leukocyte antigen. On the basis of tissue distribution and biochemical characteristics, this antigen is equine MHC II. The equine MHC II antigen was found on a large subset of T-lymphocytes in addition to all B-lymphocytes, as has been reported previously. In addition MHC II was found to be present on a large proportion of both the mutually exclusive equine T-lymphocyte subpopulations which express either the equine homologues of CD4, or CD8. In a study of changes in equine MHC II expression wi...
DelVento VR, Amann RP, Trotter GW, Veeramachaneni DN, Squires EL.A sample of testicular parenchymal tissue, approximately 2 x 7 x 7 mm, was aseptically removed from 1 testis in each of 9 stallions on day 0. Slight to moderate hemorrhage from the tunica albuginea was observed in 8 stallions, but bleeding from the parenchyma was detected in only 2 stallions. Stallions were castrated 27 days later. Normal development of granulation tissue was evident at the biopsy site, but hematomas were not observed. In situ measurement of the widths of the right and left testes, total scrotal width, and evaluation of testicular echogenicity during ultrasonography were varia...
Whithair KJ, Bowersock TL, Blevins WE, Fessler JF, White MR, Van Sickle DC.Septic arthritis was induced in one antebrachiocarpal joint of seven horses by the intra-articular injection of 1 mL Staphylococcus aureus suspension containing a mean of 10(5) colony-forming units. Twenty-four hours after inoculation, four horses were treated by regional perfusion with 1 g of gentamicin sulfate, and three horses received 2.2 mg/kg gentamicin sulfate intravenously (IV) every 6 hours. Synovial fluid was collected for culture and cytology at regular intervals, and the synovial membranes were collected for culture and histologic examination at euthanasia 24 hours after the first ...
Baumann U, Bode W, Huber R, Travis J, Potempa J.The crystal structure of active-site cleaved equine leucocyte elastase inhibitor, a member of the serpin superfamily, has been solved and refined to a crystallographic R-factor of 17.6% at 1.95 A resolution. Despite being an intracellular inhibitor with rather low sequence homology of 30% to human alpha 1-antichymotrypsin and alpha 1-proteinase inhibitor, the three-dimensional structures are very similar, with deviations only at the sites of insertions and few mobile secondary structure elements. The better resolution in comparison with the structures of other cleaved serpins allows a more pre...
Wong CW, Smith SE, Thong YH, Opdebeeck JP, Thornton JR.Chemotactic locomotion and luminol-dependent chemiluminescence of neutrophils, mitogen-induced lymphocyte blastogenesis, serum cortisol concentration, immunoglobulin quantification, and leukocyte counts were determined to evaluate the effect of a single strenuous exercise in horses. Increased serum cortisol concentration (P less than 0.01) and an increased neutrophil-to-lymphocyte ratio (P less than 0.05) indicated that horses had been stressed. The chemotactic index and peak chemiluminescence production decreased significantly (P less than 0.05 and P less than 0.01, respectively) 1 day after ...
Furr MO, Crisman MV, Robertson J, Barta O, Swecker WS.Immune system dysfunction and immunoglobulin deficiency was diagnosed in a 2-year-old horse with disseminated lymphosarcoma. Prolonged (35 days) parenteral nutrition was delivered to support the horse during a period in which immune function studies could be performed. Correction of nutritional compromise by use of parenteral nutrition did not correct the immunoglobulin deficiency, and results of lymphocyte phenotype testing did not indicate abnormal proportions of leukocytes. Lymphoblast transformation studies were suggestive of a circulating immunosuppressive factor in the horse's serum. Nor...
Magdesian KG, Brook D, Wickler SJ.The temporal response of blood and serum proteins to chronic plasmapheresis was determined in 8 horses used in a commercial antibody enterprise. Plasmapheresis of between 4 and 11 L induced significant decreases in total protein, albumin, and IgG values. With the exception of a high hematocrit value for the first postplasmapheresis blood sample, there were no changes in erythrocyte or leukocyte measurements, and no changes in the proportions of serum protein in an electrophoretic profile. Regression equations generated for recovery of proteins after plasmapheresis indicated a return to preplas...
Klei TR, Chapman MR, Dennis VA.The adherence of equine leukocytes to Strongylus vulgaris infective larvae (L3) in the presence of normal and immune sera was examined in vitro. Immune sera promoted adherence of buffy coat cells from ponies with S. vulgaris-induced eosinophilia (eosinophilic ponies) to S. vulgaris L3. However, eosinophils in the buffy coat cells were the predominant adherent cell type. Studies using leukocyte populations enriched for eosinophils, neutrophils, and mononuclear cells from eosinophilic ponies support the observations using buffy coat cells that eosinophils were the main effector cells. Adherent e...
Chong YC, Duffus WP, Hannant D.Peripheral blood mononuclear cells (PBMC) from an adult horse and from foals demonstrated natural killer (NK)-type cytotoxicity against a range of xenogeneic and allogeneic cell targets. The human tumour cell line, Chang liver was consistently the most susceptible. Chang liver, rabbit kidney (RK-13), equine sarcoid (ES) and embryonic equine kidney (EEK) cells were more susceptible when presented to horse PBMC than monolayer cultures. Embryonic equine lung (EEL) and murine YAC-1 cells conversely, were more susceptible in a trypsinized state. Horse PBMC demonstrated higher levels of NK-type acti...
Daniel GB, Tucker RL, Buckman T, Daniel SL.Isolated equine granulocytes (WBC), radiolabeled with 99mTc-hexamethylpropyleneamine oxime (99mTc-HMPAO) or 111In-oxine, were evaluated in vitro for their labeling characteristics, viability, and phagocytic function over a 6-hour postlabeling period. Mean +/- SD labeling efficiency for 111In-oxine-WBC was 62.2 +/- 15.3%, which was significantly (P less than 0.001) higher than that for 99mTc-HMPAO-WBC (32.0 +/- 17.0%). In vitro elution of radiolabel from cells was significantly (P less than 0.02) greater for 99mTc-HMPAO-WBC at 0.5, 2, and 4 hours, but was not significantly different from elutio...
Dennis VA, Klei TR, Miller MA, Chapman MR, McClure JR.Ten helminth-free pony foals divided into three groups were used in this study. Eight foals were each experimentally infected per os with 50 Strongylus vulgaris infective larvae weekly for 4 weeks, at which time one foal died of acute verminous arteritis. The remaining seven foals subsequently received 50 S. vulgaris infective larvae every 2 weeks for an additional 20 weeks. Four of the infected foals remained untreated (Group 1) and three of the infected foals were given ivermectin at 8, 16 and 24 weeks post initial infection (Group 2). Two foals served as controls (Group 3). Foals in Group 1...
Bochsler PN, Slauson DO, Neilsen NR.Neutrophil (PMN) contributions to the acute inflammatory process and host defense include generation of bioreactive oxygen metabolites and secretion of granule enzymes. We assessed equine PMN secretion using several PMN stimuli, singly and in combination with bacterial lipopolysaccharide (LPS). LPS avidly associated with equine PMN, as shown by strong PMN labeling with FITC-conjugated LPS. LPS alone (1 or 10 micrograms ml-1) was a weak stimulus for PMN superoxide anion (O2-) generation, but preincubation with LPS followed by phorbol ester (PMA, 10 ng ml-1) significantly augmented (P less than ...
Welch HM, Bridges CG, Lyon AM, Griffiths L, Edington N.The polymerase chain reaction (PCR) and co-cultivation were used to identify the lymphoreticular system as the site of latency of equid herpesvirus I (EHV-1). Primers for PCR were designed from aligned nucleotide sequences of the glycoprotein gB genes to amplify the same region of both the EHV-1 and EHV-4 genomes. Subsequent restriction digests using specific enzymes distinguished the amplified fragments of the EHV-1 genome from those of the EHV-4 genome. Ten weeks following an experimental infection of five ponies with EHV-1, latent virus was detected by PCR and recovered by co-cultivation, p...
Jaeschke G, Rudolph R.In a review supplementary to prior evaluations further hematological or cytodiagnostic aspects respectively of equine leucosis are discussed. Leukemic nature of the disease is characterized by increased count of normal blood cells or by the presence of morphologically non-differentiated cells in circulating blood or by disorder or loss of function of blood cells respectively. Thrombocytopenia in the most cases is accompanied by anemia, leukemia and tumor cell infiltration in bone marrow. Tumor cells in circulating blood indicate their presence in bone marrow too. In cases of equine leucosis wi...
Aoki T, Kimura Y, Oya A, Chiba A, Ishii M, Nambo Y.Heavy draft mares potentially have a higher risk of suffering from postpartum fever (PF) than light breed mares. The purpose of this study was to compare hematological and biochemical features between clinically healthy mares (n=40) and PF-affected mares (n=16). Mares were classified as having PF when their rectal temperature rose to >38.5°C within 96 hr of foaling. The numbers of leukocytes, lymphocytes, and neutrophils and the serum magnesium level were significantly lower in PF-affected mares. The serum SAA and NEFA levels were significantly higher in PF-affected mares. Strong inflammation...
Jaworska J, Ropka-Molik K, Kowalczyk-Zięba I, Boruszewska D, Wocławek-Potocka I, Siemieniuch M.Physiological parturition is characterized by sterile, inflammatory-like processes. During parturition, the placenta expresses various proinflammatory mediators, such as chemokines and IL-17. Nevertheless, inflammatory processes present in the parturient mare are poorly characterized. The aim of this study was to investigate the expression of selected chemokines and IL-17 in the allantochorion and the endometrium of mares that retained fetal membranes (RFM) and expelled them physiologically. We hypothesized that the expression of these mediators may be altered in the placenta of mares with RFM...
Riggs LM, Krunkosky TM, Noschka E, Boozer LA, Moore JN, Robertson TP, Peroni JF.To compare characteristics and enzymatic products of leukocytes detected in the skin and laminar tissues of horses administered black walnut heartwood extract (BWHE) and horses administered purified lipopolysaccharide (LPS). Methods: 25 healthy 5- to 15-year-old horses. Methods: Horses were randomly assigned to receive LPS (20 ng of O55:B5 Escherichia coli endotoxin/kg; n = 5) IV or 6 L of BWHE (10) or water (control group; 10) via nasogastric intubation. Horses were euthanatized 12 hours after treatment or at onset of Obel grade 1 lameness. Laminar tissue samples and skin samples from the mid...
Lindberg A, Näsman-Glaser B, Lindgren JA, Robinson NE.To evaluate leukotriene (LT) biosynthetic capacity in lung tissue from healthy horses and horses with recurrent airway obstruction (RAO). Methods: Lung parenchyma and airway specimens from 8 RAO-affected and 5 healthy horses. Methods: Horses were stabled for > or = 72 hours. Blood was drawn before euthanasia, after which lung specimens were collected. Tissue strips from small airways and parenchyma were incubated in organ baths with the precursor LTA4 or stimulated with calcium ionophore A23187 or the tripeptide N-formyl-Met-Leu-Phe (fMLP), with or without exogenous arachidonic acid, in the...
Weiss DJ, Evanson OA.To determine the potential usefulness of tests for detection of platelet activation and platelet-leukocyte aggregates in horses. Methods: Blood from 3 healthy Thoroughbreds. Methods: Microscopic and flow cytometric assays were used to evaluate spontaneous platelet aggregation, platelet activation, and platelet-leukocyte aggregates. Platelet activation was detected by evaluation of binding of anti-human fibrinogen to unactivated and ADP-, thrombin-, thrombin agonist receptor peptide-, and platelet activating factor-activated platelets. Platelet-leukocyte aggregates were evaluated microscopicall...
Frauenfelder HC, Fessler JF, Moore AB, Bottoms GD, Boon GD.The effects of dexamethasone (1 mg/kg of body weight) on hematologic, blood gas, and blood coagulation values in anesthetized ponies during endotoxin-induced shock were evaluated. Fifteen ponies were assigned to 3 groups of 5 ponies each: group 1, anesthetized nontreated and dexamethasone-treated controls; group 2, endotoxin, nontreated; group 3, endotoxin, dexamethasone treated. The hematologic changes in this endotoxin shock model included leukopenia and hemoconcentration. Significant hematologic effects were not seen in ponies after administration of dexamethasone. However, dexamethasone tr...
Escribano BM, Castejón FM, Vivo R, Agüera S, Agüera EI, Rubio MD.The aim of the present paper was: (1) to find out if there were any differences in the nonspecific immunological pattern of peripheral blood neutrophil between two breeds of horses (AA and SA); (2) to evaluate the effects of an exercise in the aerobic-anaerobic threshold. This has been observed in a group of 11 untrained horses (6 SA and 5 AA) of 2.5 years old. No statistically significant differences were found in the different stages of immune response between the rest and immediately after physical exercise to two breeds. However, the chemotaxis was significant higher at rest in the AA than...
Williams JM, Lin YJ, Loftus JP, Faleiros RR, Peroni JF, Hubbell JA, Ravis WR, Belknap JK.Laminitis is a serious complication of horses suffering from sepsis/endotoxaemia-related events. Laminitis in horses and organ injury in human sepsis are both reported to involve inflammatory injury to the laminae/organs including early activation of endothelium and leucocytes leading to emigration of neutrophils into the tissue interstitium. In the black walnut extract (BWE) model, systemic inflammatory events coincide with marked increase in laminar mRNA concentrations of inflammatory genes including proinflammatory cytokines (i.e. IL-1beta, IL-6), COX-2, chemokines (i.e. IL-8) and endotheli...
Tosi I, Bureau F, Farnir F, Denoix JM, Lekeux P, Art T.Cytosine-phosphate-guanosine (CpG-ODN) has been described as a potent immunostimulatory agent in different species. No study reported the effect of a P-class CpG when administered systemically in healthy horses. The aim of this study was to evaluate the tolerance and the effect of an intramuscularly administered P-class CpG-ODN on hematology and on plasma cytokines (IFN-α, IL-10, TNF-α, IFN-γ) in 8 healthy horses. Intra-muscular CpG-ODN or placebo (PBS) was administered twice at a 7 days-interval. Groups were inversed after 2 months of washout period. A physical examination, complete bloo...
Alberghina D, Statelli A, Monteverde V, Vazzana I, Cascone G, Panzera M.The values for basal serum cortisol concentrations of horses are available in many studies. However, there are limited data about serum cortisol in donkeys. The present study aimed to determine the baseline values for serum cortisol, to evaluate the influence of age and pregnancy on its levels, and to correlate its values with leucocyte profile, serum cholesterol, and triglycerides. Serum samples were collected from 97 healthy donkeys. Cortisol was analyzed by chemo-luminescent assay. The median and the 2.5th and 97.5th percentiles of serum cortisol measured and calculated in all donkeys were ...
Treonze KM, Alves K, Fischer P, Hagmann WK, Hora D, Kulick A, Vakerich K, Smith ND, Lingham RB, Maniar S, Reger TS, Zunic J, Munoz B, Prasit P....The purpose of this study was to characterize the alpha(4)beta(1) receptor (CD49d/CD29, very late antigen-4, VLA-4) on circulating equine leukocytes and to evaluate the intrinsic potency of an alpha(4)beta(1) receptor antagonist (Compound B) in the horse. Ultimately, these studies would allow us to determine the suitability of treating recurrent airway obstruction (RAO; heaves) affected horses by blocking the cellular recruitment of lymphocytes and neutrophils into the lung. The data demonstrates the alpha(4)beta(1) integrin is present on horse lymphocytes and neutrophils (fluorescence-assiste...
Wilks CR, Barton MD, Allison JF.Immune responses to Rhodococcus equi were assayed in mares and foals on 7 studs in south-eastern Australia using skin test reactivity to the intradermal injection of culture filtrate and an indirect fluorescent antibody test. The prevalence of positive skin-test reactions did not differ between studs with a history of R. equi disease and those without but there were more mares with high antibody titres on studs with a disease history. A leucocyte extract prepared from mares that were skin-test positive was evaluated for its ability to protect foals exposed to experimental or natural challenge:...
Turek JJ, Templeton CB, Bottoms GD, Fessler JF.Endotoxic shock was induced in 5 ponies by intraperitoneal injections of 20, 40, 60, 80, and 80 micrograms of Escherichia coli endotoxin (LPS)/kg of body weight at 0, 6, 12, 18, and 24 hours, respectively. At 24 hours, the ponies also were given 20 micrograms of LPS/kg via catheter in the left ventricle of the heart. A 2nd group of 4 ponies was given 1.1 mg of flunixin meglumine (FM)/kg, IV, at 6, 12, 18, and 24 hours just before the corresponding LPS injection. Two hours after the 24-hour LPS injection, the ponies in both groups were anesthetized, the lungs were perfused with fixative, and po...
Bailey SR, Elliott J.Endotoxin has been implicated in the pathophysiology of acute laminitis. The aim of this study was to examine the direct effects of endotoxin on isolated equine digital blood vessels. Equine digital veins (EDV), incubated in Krebs-Henseleit solution containing lipopolysaccharide (LPS) (1 microg/ml) became hyporesponsive to 5-HT after 16 h. Cycloheximide and ibuprofen blocked this effect of LPS and increased the maximum response obtained to 5-HT when compared to control vessels. L-nitroarginine methyl ester (L-NAME) reversed the hyporesponsiveness caused by LPS. Vessels maintained in culture me...
Perryman LE, McGuire TC.Combined immunodeficiency in horses is a genetic disorder in which there is a defect in the production of committed B and T lymphocytes. In this study, peripheral blood mononuclear leukocytes from foals with combined immunodeficiency were examined for their capacity to stimulate and respond in one-way mixed lymphocyte cultures. Irradiated cells from combined immunodeficient foals were uniformly capable of stimulating cells from unrelated horses. However, none were able to respond to allogeneic stimulation. Examination of cells from known carrier horses revealed no difference in capacity to sti...
Wotman KL, Chow L, Martabano B, Pezzanite LM, Dow S.Ocular surface squamous neoplasia (OSSN) is the major cause of corneal cancer in man and horses worldwide, and the prevalence of OSSN is increasing due to greater UVB exposure globally. Currently, there are no approved treatments for OSSN in either species, and most patients are managed with surgical excision or off-label treatment with locally injected interferon alpha, or topically applied cytotoxic drugs such as mitomycin C. A more broadly effective and readily applied immunotherapy could exert a significant impact on management of OSSN worldwide. We therefore evaluated the effectiveness of...
Hosoya M, Inoue A, Kimura N, Arai T.Glucose, free fatty acid (FFA), triglyceride (TG) and immunoreactive insulin (IRI) concentrations in plasma and activities of enzymes related to energy metabolism in some types of peripheral leukocytes were measured in thoroughbred race horses before and after racing. Glucose, FFA, TG and IRI concentrations and enzyme activities did not change significantly in plasma. However, the activities of cytosolic hexokinase, malate dehydrogenase (MDH), mitochondrial MDH and aspartate aminotransferase decreased significantly in leukocytes of the horses after the races. The cytosolic ratio of MDH/LDH act...
Meichner K, Kraszeski BH, Durrant JR, Grindem CB, Breuhaus BA, Moore PF, Neel JA, Linder KE, Borst LB, Fogle JE, Tarigo JL.An 11-year-old, 443-kg Haflinger mare was presented to the North Carolina State University Veterinary Teaching Hospital with a 2-week history of lethargy and a 3-day duration of anorexia, pyrexia, tachycardia, and ventral edema. Severe pitting edema, peripheral lymphadenopathy, and a caudal abdominal mass were noted on physical examination. An extreme leukocytosis (154.3 × 103 /μL) and microscopic hematologic findings suggestive of myelomonocytic leukemia were observed. Serum protein electrophoresis revealed a monoclonal gammopathy and urine protein electrophoresis revealed a monoclonal ligh...
Flaminio MJ, Rush BR, Davis EG, Hennessy K, Shuman W, Wilkerson MJ.This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst,...
Brudvig JM, Swenson CL.Rapid and precise measurement of total and differential nucleated cell counts is a crucial diagnostic component of cavitary and synovial fluid analyses. Objective: The objectives of this study included (1) evaluation of reliability and precision of canine and equine fluid total nucleated cell count (TNCC) determined by the benchtop Abaxis VetScan HM5, in comparison with the automated reference instruments ADVIA 120 and the scil Vet abc, respectively, and (2) comparison of automated with manual canine differential nucleated cell counts. Methods: The TNCC and differential counts in canine pleura...
Lindberg A, Robinson NE, Näsman-Glaser B, Jensen-Waern M, Lindgren JA.To determine the ex vivo leukotriene (LT) biosynthesis in peripheral blood neutrophils (PBNs) and inflammatory cells in bronchoalveolar lavage fluid (BALF) obtained from horses affected with recurrent airway obstruction (RAO). Methods: 6 RAO-affected and 6 control horses. Methods: Before and 6, 24, and 48 hours after stabling, disease severity was determined subjectively by clinical and mucus scores and measurement of the maximal change in pleural pressure (deltaPpl(max)); PBNs were isolated and BALF samples were examined cytologically. The PBN and BALF cells were activated with a calcium iono...
Walters M, Skovgaard K, Andersen PH, Heegaard PMH, Jacobsen S.The role of resident cells such a synoviocytes and chondrocytes in intra-articular inflammation is well-characterized, however the in vivo gene expression patterns of cells (predominantly leukocytes) in the synovial fluid (SF) of an inflamed joint have never previously been investigated. The aim of this study was to investigate gene expression in SF leukocytes from the inflamed joint cavity after intra-articular lipopolysaccharide (LPS) injection in horses to improve our understanding of the temporal regulation of the intra-articular inflammatory response. Gene expression was investigated in S...
Yager JA, Duder CK, Prescott JF, Zink MC.Polymorphonuclear neutrophil leukocytes (PMNL) from 8 healthy foals (2-14 weeks of age) and 2 foals with bacterial pneumonia were separated from whole blood using a 2 step Percoll gradient. Purified PMNL were tested for bactericidal function against Rhodococcus equi and Staphylococcus aureus in the presence of normal horse serum. The percentage uptake after a 15-min pre-incubation of PMNL and bacteria was also calculated. Ultrastructural examination of the interaction of R. equi and normal foal PMNL was performed after 15 min incubation. Results indicated that foal PMNL effectively phagocytose...
Janda J, Lehmann M, Luttmann W, Marti E.Interleukin-3 is a growth and differentiation factor for various hematopoietic cells. IL-3 also enhances stimulus-dependent release of mediators and cytokine production by mature basophils. Function of IL-3 has not been studied in horses because of lack of horse-specific reagents. Our aim was to produce recombinant equine IL-3 and test its effect on sulfidoleukotriene and cytokine production by equine peripheral blood leukocytes (PBL). Equine IL-3 was cloned, expressed in E. coli and purified. PBL of 19 healthy and 20 insect bite hypersensitivity (IBH)-affected horses were stimulated with Culi...
Vallejo AN, Miller NW, Clem LW.This work was undertaken to investigate whether or not antigen processing and presentation are important in channel catfish in vitro secondary immune responses elicited with structurally defined proteins, namely, pigeon heart cytochrome C (pCytC), hen egg lysozyme, and horse myoglobin. The use of in vitro antigen-pulsed and fixed B cells or monocytes as antigen presenting cells (APC) resulted in autologous peripheral blood leukocytes (PBL) responding with vigorous proliferation and antibody production in vitro. In addition, several long-term catfish monocyte lines have been found to function a...
Watson ED, Stokes CR, Bourne FJ.Both random and directional migration of blood neutrophils from 9 mares susceptible to persistent endometritis were significantly less (p less than 0.05) than neutrophils from 8 resistant mares. Serum from susceptible mares had significantly more (p less than 0.01) chemotactic activity than serum from resistant mares. Although phagocytosis of yeast blastospores by blood neutrophils from 4 resistant and 3 susceptible mares was similar, uterine neutrophils from susceptible mares were significantly worse (p less than 0.01) at phagocytosis than uterine neutrophils from resistant mares. Uterine was...
Kimura S, McCoy AM, Boothe DM, Wooldridge AA, Graff E, Hammack SM, Cresci L, Hofmeister E, Lascola KM.To describe misoprostol pharmacokinetics and anti-inflammatory efficacy when administered orally or per rectum in endotoxin-challenged horses. Methods: 6 healthy geldings. Methods: A randomized 3-treatment crossover design was performed with a minimum washout period of 28 days between treatment arms. Prior to endotoxin challenge (lipopolysaccharide, 30 ng/kg IV over 30 minutes), horses received misoprostol (5 µg/kg once) per os (M-PO) or per rectum (M-PR) or water as control (CON). Clinical parameters were evaluated and blood samples obtained to measure plasma misoprostol free acid concentrat...
Grondin TM, DeWitt SF, Keeton KS.A 9-year-old Arabian mare was evaluated for a 7-day history of malaise. Results of a CBC included a leukocyte concentration within the reference interval (8.4 x 10(3)/microL, reference interval 6.0-14.0 x 10(3)/microL) with an apparent degenerative left shift (segmented neutrophils 1.2 x 10(3)/microL, reference interval 2.5-7.5 x 10(3)/microL; hyposegmented neutrophils 1.8 x 10(3)/microL, reference interval 0.0-0.2 x 10(3)/microL). Serum clinical chemistry results included increased aspartate transaminase, alkaline phosphatase, and gamma-glutamyltransferase activities. A presumptive diagnosis ...
Mérant C, Bonnefont C, Desbos A, Greenland T, Cadoré JL, Monier JC.The recognition of equine lymphocyte antigens by monoclonal antibodies (mAbs) directed against human CD11a, CD18, CD21, CD23, CD29 and DR, as well as mouse CD23 was studied by flow cytometry. Unlike anti-CD11a, -CD21, -CD23 and DR mAbs, anti-CD18 and CD29 mAbs labelled the same percentage of horse peripheral blood lymphocytes (PBL) as human PBL. Double-staining with anti-horse immunoglobulin antibodies showed that anti-CD21 and -CD23 mAbs are mainly bound to peripheral blood B lymphocytes. The seven mAbs were also tested on the lymph node and thymus cells. The molecular targets of anti-CD11a, ...
Hansen PJ, Hoggard MP, Rathwell AC.The ability of stallion seminal plasma to modify phagocytosis of spermatozoa and Streptococcus zooepidemicus was examined. Phagocytosis was monitored indirectly as the H2O2 produced by peripheral blood leukocytes after addition of spermatozoa or bacteria. Hydrogen peroxide production after addition of ejaculated spermatozoa was greater (P less than 0.01) than after addition of epididymal sperm. Furthermore, pre-incubation of epididymal sperm with 6.25-50% seminal plasma caused a dose-dependent increase in subsequent H2O2 production by leukocytes (P less than 0.05). In addition, equine serum wa...
