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.
Zinkl JG, Brown PD.Horse polymorphonuclear leukocytes (PMN) isolated from horse blood by sedimentation and isotonic lysis and having about 25% accompanying lymphocytes were as effective at chemotaxis as nearly pure PMN isolated by density gradient techniques. N-Formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP), used as a representative of the formylmethionyl peptides (produced by prokaryocytic organisms), was effective as a chemoattractant only at the high concentration of 10(-4) M. When serum was preincubated with FMLP at concentrations as low as 10(-8) M, the serum attracted horse PMN. This activity was not g...
Wagner AE, McIlwraith CW, Martin GS.Orgotein was injected into the right intercarpal joint of each of 8 horses; the corresponding left joint was left alone (not injected) or was given an injection of normal saline solution. Injection with orgotein caused a transient, marked inflammatory response, evidenced by clinical signs and by increased leukocytes and total protein in the synovia (synovial fluid). Leukocyte numbers and total protein concentration were increased (P less than 0.010) in the orgotein-injected joints within 24 hours. However, saline solution alone also elicited a marked inflammatory response, manifested by increa...
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...
Brumbaugh GW, Stitzel KA, Zinkl JG, Feldman BF.Myelomonocytic myeloproliferative disease in a horse was diagnosed on the basis of hematologic, enzymatic, and histopathologic findings. It was characterized clinically by depression, weight loss splenomegaly, lymphadenopathy, coagulopathy, and bacteremia. Hematologic findings included severe refractory anemia, thrombocytopenia, monocytosis, and pleomorphic leukocytes, with a left shift of the myeloid series. The serum lysozyme concentration was 14.5 microgram/ml (normal, less than 5 microgram/ml). The bone marrow contained many immature cells of the myeloid series and had a myeloid-to-erythro...
Jörg A, Henderson WR, Murphy RC, Klebanoff SJ.Horse eosinophils purified to greater than 98% generated slow reacting substance (SRS) when incubated with the calcium ionophore A23187. On a per cell basis, eosinophils generated four to five times the SRS produced by similarly treated horse neutrophils. Eosinophil SRS production was inhibited by 5,8,11,14-eicosatetraynoic acid and augmented by indomethacin and arachidonic acid, suggesting that it was a product(s) of the lipoxygenase pathway of arachidonic acid metabolism. Compounds with SRS activity were purified by high-pressure liquid chromatography (HPLC) and identified by ultraviolet spe...
Wagner AE, Bennett DG.Eighteen clinically normal horses were used to study the characteristics of normal thoracic fluid. Thoracic fluid was obtained from each horse and was found to be similar to equine abdominal fluid. Total leukocytes averaged 3994/ul, total protein 1.8 g/dl, and specific gravity 1.015. Analysis of thoracic fluid from 16 horses with clinical signs of thoracic disease showed abnormalities in every case. Thoracic fluid analysis alone determined a specific diagnosis in 50% of the cases.
Heyneman RA, Vercauteren RE.The subcellular components of purified neutrophil leukocytes from horse blood were fractionated by isopyknic equilibration in sucrose and metrizamide gradients. Five classes of particles have been identified: dense azurophil granules containing the bulk of the lysosomal acid hydrolase and peroxidase activity (A); less dense particles, containing all the lysozyme activity, but not resolved from a second population of azurophils B, and particles of low density, biochemically characterized as a plasma membrane fraction (C). Isopyknic equilibration in sucrose disclosed a minor membrane fraction (D...
Blue MG, Brady AA, Davidson JN, Kenney RM.Forty uterine fluid samples were obtained during oestrus and dioestrus of successive cycles from 4 mares classified as resistant to bacterial infection of the uterus. The flushings were assayed for peroxidase activity and ability to influence phagocytosis by polymorphonuclear neutrophils. Uterine fluid volumes and protein concentrations and serum oestrogen values were similar during oestrus and dioestrus. Serum progesterone concentrations were depressed during oestrus. Equine neutrophils adherent to glass slides were exposed to a genital strain of Streptococcus zooepidemicus suspended in salin...
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:...
Potempa J.The proteolytic activity of the elastase-like proteinase from granules of horse blood leukocytes is retained on a column of Cibacron Blue-Sepharose and can be eluted with 0.5 M KSCN. During this procedure its mol. wt. is reduced from 49000 to 30000 and isoelectric point is shifted towards higher pH. The inactive protein not adsorbed on Cibacron Blue-Sepharose is strongly acidic and shows a mol. wt. of 20000. Upon mixing this protein with the modified enzyme the native proteinase is reconstituted as shown by polyacrylamide gel electrophoresis at pH 8.3 and isoelectric focusing in a sucrose grad...
Moore JN, Garner HE, Coffman JR.Experimental carbohydrate overload of 15 horses resulted in consistent haematological changes during development of laminitis hypertension. Significant alterations in packed cell volume, leucocyte differential count, serum glucose levels and protein values occurred before onset of Obel grade 3 lameness and hyperkinetic circulatory state. Blood platelets were significantly decreased 8 h after the onset of severe lameness. Findings in this study were indicative of haemoconcentration due to compartmental fluid shifts and leucocytic stress response consistent with increased circulating adrenogluco...
Banks KL, Greenlee A.Various cell populations of equine mononuclear leukocytes were identified and isolated. Mononuclear leukocytes were concentrated by isopyknic centrifugation, using a solution of Ficoll and Hypaque. Three additional techniques were explored to separate monocytes from lymphocytes, and 3 methods were used to separate lymphocyte types. Cytochemical techniques for the detection of nonspecific esterase readily distinguished equine monocytes from lymphocytes. Peripheral blood lymphocytes were separated into at least 2 populations. One population had surface traits identical to thymocytes [ie, they re...
Dutta SK, Bumgardner MK, Scott JC, Myrup AC.Various methods of separation and identification of major equine leukocyte populations and subpopulations were used. The purity of T and B lymphocytes separated in Sephadex anti-equine F(ab')2 columns was 87% to 99% and 83% of 97%, respectively. The purity of T lymphocytes separated in nylon-wool columns was 89% to 98%. Preparations of B lymphocytes separated in glass-bead columns were 68% to 79% pure. The presence (or absence) of surface immunoglobulin by immunofluorescence was the most consistent and reliable method for the identification of B or T lymphocytes, respectively. However, the ery...
Searcy GP, Orr JP.A nine year old quarter horse exhibited progressive weight loss and inappetance over a 47 day period. There was clinical evidence of pleuritis and pneumonia substantiated by leukocytosis and elevated protein in pleural fluid. Over the entire period the horse was neutropenic and had circulating abnormal immature granulocytes and low numbers of blast cells. Anemia and thrombocytopenia progressively worsened. Bone marrow examination revealed very few mature granulocytes but large numbers of immature cells of the granulocytic series and marked megaloblastic transformation of erythroid cells. These...
Brownlow MA, Hutchins DR, Johnston KG.Twenty horses, aged one to 17 years (mean age 6 years), presented for elective destruction and subsequently found at autopsy to have no significant peritoneal alterations, were used to determine a variety of reference values for peritoneal fluid. Samples were collected ante mortem or within 1 h post mortem. Each cavity contained 100 to 300 ml of usually clear, pale yellow fluid which in a clinical refractometer showed a mean specific gravity 1.010 (range 1.0081-1.0116) and mean (+/- standard deviation) total protein 7.7 +/- 3.6 g/litre. The mean total nucleated cell count (+/- sd) was 4.33 +/-...
Carakostas MC, Moore WE, Smith JE.A procedure with chromium-51 (51Cr) as the cell label that maintains high-cell viability for studying granulocyte kinetics in horses is described. The procedure combines and modifies several methods for isolating leukocytes and granulocytes for use in the horse when a large volume of labeled cells is required. Also described is an improved technique for measuring granulocyte specific activity in large serial blood samples, using a Ficoll-sedimentation method. The procedure should be useful for determining granulocyte kinetics in the horse, the only major domestic species for which such data ar...
Carakostas MC, Moore WE, Smith JE, Johnson D.The functional capacity of the marrow granulocyte reserve (MGR) in 4 adult horses was studied, using 51Cr-labeled leukocytes. The mean increase in the peripheral granulocyte count following injections of etiocholanolone (0.3 mg/kg) was 870 granulocytes/mm3, and the mean increase following prednisolone administration (200 mg) was 5,880 granulocytes/mm3. Etiocholanolone failed to mobilize the MGR and decreased the rate of granulocyte egress from the blood. Prednisolone rapidly mobilized the MGR and markedly decreased the granulocyte specific activity during the first 3 hours after injection.
Allen BV.A technique for automating equine differential leucocyte counts by analysis of volume distribution curves using the Coulter Channelyzer has been developed and evaluated. A comparison between the results obtained by this method and standard microscopic techniques showed good agreement in most cases. Blood samples can be analysed for both differential and total leucocyte counts at a rate of 25/h. For each sample an average 16,000 leucocytes are classified by the Channelyzer. The method of volume analysis is suitable for the precise counting of polymorphonuclear neutrophils, lymphocytes and eosin...
Koj A, Kurdowska A.Antithrombin III and alpha 1-proteinase inhibitor isolated simultaneously from horse citrated plasma were tested for inhibitory activity against bovine trypsin and chymotrypsin, as well as elastase-like neutral proteinases from horse leucocytes. The stoichiometry of reaction and kinetic parameters (kass, Ko) were estimated and related to the protein pattern obtained after exposure of these proteinases to horse inhibitors as analyzed by polyacrylamide gel electrophoresis (PAGE and PAGE-SDS). As shown by fast reaction rates and low values of dissociation constants the two inhibitors effectively ...
Gleeson LJ, Coggins L.Twenty-one pregnant mares were inoculated with EHV1. Nineteen became infected as evidenced by clinical signs and/or viremia but only one mare aborted a virus-infected fetus. The viremias were leukocyte-associated and appeared to be non-productive, latent infections of these cells. Infectivity, detectable by cocultivation, persisted in the circulating leukocytes for as long as 9 days without resulting in abortion. The data suggest that it is extremely difficult to evaluate the efficacy of vaccines in preventing EHV1 (Rhinopneumonitis) abortion due to the paucity of non-exposed mares, lack of te...
Lazary S, Bullen S, Müller J, Kovacs G, Bodo I, Hockenjos P, De Weck AL.Mono- and oligospecific lymphocytotoxic alloantibodies from primiparous mares were tested on cells from horse families of various breeds in the two-step microcytotoxicity assay. The results showed that the detected antigens were inherited co-dominantly and autosomally as simple Mendelian traits. The membrane antigens showed different linkage with one or more other antigens and seem to be coded by a limited number of loci (at least three) from one chromosome. In the families tested one recombinant for the serologically defined antigens was recognized. The mixed leukocyte reactions of cells from...
Ferrante A, Thong YH.Results presented show that highly purified populations of mononuclear (MN) and polymorphonuclear (PMN) leucocytes can be obtained from horse blood by a procedure similar to that previously described for the separation of these leucocytes from human blood. This involved centrifugation of horse blood on a Ficoll-Hypaque medium with a density of 1.095 g/ml. The procedure required approximately 1 h for completion and resulted in the simultaneous preparation of MN (greater than 98% purity) and PMN (greater than 96% purity) leucocytes. Cell viability exceeded 95% and cells retained immunological fu...
Kings MA, de Weck AL.Pharmacological histamine releasing agents, such as compound 48/80, poly-L-lysine, adrenocorticotrophic hormone (ACTH; beta 1-24 available commercially as Synacthen), catecholamines, purine bases, etc., are well known to induce histamine release from rat peritoneal mast cells and mast cells of other species; and to a lesser extent from peripheral blood leucocytes. It is reported in this paper that several of these potent histamine-releasing agents induce little or no histamine release from horse leucocytes. In particular the calcium ionophore A 23187 induced no histamine release. On the other ...
Ek N.Studies of Pr protein concentrations in sera of sick horses were carried out using ’s (1965) immunodiffusion technique. Relative values against a chosen standard of 100 were determined for a total of 102 horses. Horses with acute infections had Pr protein values significantly above the normal. The highest individual Pr protein value recorded in this group was 202. Horses suffering from acute laminitis and malignant tumours also had increased Pr protein values. There was a positive correlation between the Pr protein value and the blood leucocyte count and a negative correlation between the P...
Froscher BG, Nagode LA.Alkaline phosphatase isoenzymes from small intestine, cecum, large colon, small colon, liver, kidney, leukocytes, and serum from ten clinically normal horses were defined by their sensitivities to L-phenylalanine, L-homoarginine, levamisole and heat, and by polyacrylamide gel disc electrophoresis. Readily identifiable isoenzymes occurred in small intestine, granulocytes, kidney, cecum, and large and small colon. By contrast, alkaline phosphatases from liver, lymphocytes, and serum could not be discriminated by this group of tests.
Torbeck RL, Prieur DJ.Cartilaginous defects were created in the radiocarpal joints of 12 horses. Synovial fluid cytologic features, lysozyme activity, and beta-glucuronidase activity were monitored for 16 days. A comparison was made of plasma lysozyme and beta-glucuronidase activity and of synovial fluid lysozyme, beta-glucuronidase, and leukocyte concentrations. Plasma lysozyme was found to be independent of synovial fluid lysozyme activity. Synovial fluid lysozyme was significantly increased (P less than 0.001) in all joints with surgically induced defects (group I) compared with controls (arthrocentesis done; gr...
Schürmann C, Loose M, Failing K, Wehrend A.The aim of this study was to analyze data concerning history, clinical course, and prognosis of retained fetal membranes in mares. Methods: Patient records of 121 hospitalized mares with retained placenta were evaluated. In 82 cases, additional blood examinations were performed and analyzed. Results: There was no significant correlation between age, parity or course of parturition and retained placenta. Eighty-one mares (66.9 %) were presented solely with retained fetal membranes, 40 mares (33.1 %) had additional diseases at presentation, most commonly injuries of the labia, followed by perine...
Laskoski LM, Locatelli-Dittrich R, Valadão CA, Deconto I, Gonçalves KA, Montiani-Ferreira F, Brum JS, de Brito HF, de Sousa RS.The present study was aimed at identifying laminar lesions and leukocyte infiltration in hoof laminar tissue of horses with colic syndrome and its correlation with the total leukocyte count before death. Six healthy horses were used as control group (CG), and eighteen horses with lethal gastrointestinal disease were divided into two groups: leukopenic group (LG) with seven leukopenic horses, and non-leukopenic group (NLG) with 11 horses with total leukocyte count within reference range for the species. Leukocyte infiltration was examined by immunohistochemistry. Laminar lesions were observed i...
Lopes MA, Salter CE, Vandenplas ML, Berghaus R, Hurley DJ, Moore JN.To investigate the effect of ex vivo exposure to lipopolysaccharide (LPS) on the expression of inflammatory genes in leukocytes from horses with gastrointestinal (Gl) disease and determine whether the pattern or magnitude of the response to LPS correlated with the type of disease and outcome. Methods: 49 horses with Gl disease and 10 healthy horses Methods: Leukocytes were isolated from blood samples and submitted to 3 protocols: immediate freezing, freezing after 4-hour incubation in medium, and freezing after 4-hour incubation in medium containing LPS. Expression of 14 genes associated with ...
Feige K, Ehrat FB, Kästner SB, Schwarzwald CC.The purpose of this study was to evaluate plasmapheresis as a method for plasma extraction in comparison with centrifugation or gravity sedimentation. The study was designed as a cross over trial with six Freiberger horses undergoing plasma donation by plasmapheresis followed by whole-blood donation and subsequent plasma production 4 weeks later. Automated plasmapheresis and whole-blood donation were well tolerated in all horses. The plasmapheresis method achieved an almost complete removal of erythrocytes and leucocytes from plasma at all flow rates. After blood bag centrifugation, significan...
Kampl B, Dominis-Kramaric M, Bacar-Huskic L.The activities of leukocyte alkaline phosphatase (l-AP) and serum gamma-glutamil transferase (gamma-GT), and total leukocyte counts were determined in horses submitted to the production of hyper immune sera against tetanus (Clostridium tetani). The purpose of this work was to investigate the prospective changes of mentioned parameters in horses under the described circumstances. In addition, the suitability of these parameters in assessing the health condition of the same horses had to be evaluated. The average total leukocyte count increased in one month from the values considered as physiolo...
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...
Kelley DE, Schnobrich MR, Gayer S, Scoggin C, Bradcamp E, Canisso IF.The objective of this study was to compare a commercially available leukocyte esterase strip test with endometrial cytology in diagnosing endometritis and establish a cutoff point, sensitivity, and specificity for the leukocyte esterase test (LET). Forty-six light breed mares presented for breeding management were enrolled in this study. Transrectal palpation and ultrasonography examinations were performed to determine when the mares were in estrus. Kalayjian endometrial swabs were used for culture, cytology, and the LET by using the cap to retrieve an endometrial scraping. Thirty-six mares ha...
Zaleska M, Anusz K, Winnicka A, Kita J.Twelve horses, all of them 10 years old, were vaccinated intramuscularly on 0 and 28 days of the experiment with inactivated vaccine containing only antigens of A-equi-2/Miami/63. Another three unvaccinated horses, each at the age of 10 years, were the negative control group. One, ten-year-old horse was vaccinated with commercial inactivated vaccine containing both antigens of A-equi-2/Miami/63 as well as A-equi-1/Praha/56 as positive control. Three horses were challenged intranasally with homotypic strain of Miami/63, while six other were challenged with heterotypic strains--three with Suffol...
Gordon BJ, Latimer KS, Murray CM, Moore JN.Continuous-flow centrifugation leukapheresis techniques were used to collect 300-ml volumes of leukocyte-rich plasma from 5 nonmedicated horses and from 5 corticosteroid-stimulated horses. White blood cell counts and differential counts were performed on the horses before (base line) and up to 48 hours after leukapheresis. Systemic administration of hydrocortisone increased numbers of total WBC and neutrophils and improved harvest of these cells. Nonmedicated horses had a mean yield of 3.38 X 10(10) leukocytes in the 300-ml volume. Stimulated horses yielded a mean of 6.88 X 10(10) leukocytes. ...
Clyne MJ.Proteoglycan degradation is central to the development of degenerative joint disease. Proteoglycans may be degraded by lysosomal enzymes from chondrocytes, synoviocytes or leucocytes. Collagen and matrix degradation occurs either by direct damage or due to degrading enzymes released into synovial fluid. Once the pathological sequence has begun it continues in a cyclic manner unless arrested by the ability of chondrocytes to synthesise sufficient matrix components. Treatment should ideally be directed to this end.
Escribano BM, Castejón FM, Santisteban R, Agüera EI, Tovar P, Vivo R, Rubio MD.Gender differences have not been shown in relation to the immune system in athletic horses. The aim of the present paper was to elucidate gender differences in the non-specific immune response of the polymorphonuclear neutrophil (PMN), plasma glucose and in hormones before and after an exercise in the lactate threshold (LT). A group of 12 Anglo-Arabian horses (6 females and 6 males, 4-7 years old) was observed. A submaximal exercise test was carried out at the LT. The results showed that males had a higher PMN percentage, plasma glucose values, Adherence index (AI) and random migration than fe...
Levine RA, Hart AH, Wardlaw SC.Using quantitative buffy coat analysis (QBCA), rapid and accurate measurements can be made of the erythrocyte PCV, total WBC count, and platelet count, and the leukocyte population can be differentiated into total granulocytes (including quantitation of eosinophils), and lymphocytes and monocytes. The QBCA is performed by placing a blood sample (50 to 111 microliters) into a high-precision-bore microhematocrit tube that contains a freely moving, closely fitting, cylindrical plastic float. After centrifugation for 5 minutes, the buffy coat components separate by density. The plastic cylinder fl...
Ozaydın T, Celik I, Sur E, Oznurlu Y, Uluısık D.We attempted to characterize the cytochemical staining patterns of leukocytes and to determine the percentages of alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACPase) positive lymphocytes in peripheral blood of thoroughbred foals at different ages. Blood samples were obtained from the jugular veins of 60 healthy thoroughbred foals, 1 day, 3 days and 1 year old. Each age group included 10 male and 10 female animals. Peroxidase (PO) activity was detected in neutrophils, eosinophils and monocytes. Lymphocytes were negative for PO staining. Periodic acid-Schiff (PAS) staining was o...
Allen AL, Myers SL, Searcy GP, Fretz PB.The Hematologic values of 19 equine fetuses between 202 and 238 days gestation were compared with those of their dams. The red blood cell (RBC) count, hemoglobin concentration, hematocrit, and mean corpuscular hemoglobin concentration were significantly lower in fetal blood, while the mean corpuscular volume, mean corpuscular hemoglobin, and red cell distribution width were significantly higher. Mares had a significantly higher nucleated blood cell count than fetuses, and all nucleated cells were leukocytes (WBC). Most WBC in mare blood were segmented neutrophils and lymphocytes. In contrast, ...
Weiser MG.Using a single channel electronic cell counter and attached particle size analyzer, leukocyte size distribution histograms were determined on canine, feline, bovine, and equine blood diluted with chloride-based diluent and treated with a conventional stromatolysin. Histograms were usually unimodal, but a few were bimodal. Mean values for mean lysed leukocyte particle volume were 49.2, 51.1, 55.4, and 65.0 fl for canine, feline, equine, and bovine blood, respectively. From inspection of histograms, a lower threshold of 30 fl referenced to latex spheres was interpreted to be appropriate for coun...
Marr K, Marsh K, Hernandez L, Cunningham FM, Lees P.Leukotrienes, products of the 5-lipoxygenase pathway of arachidonic acid metabolism, possess properties consistent with their involvement in a range of inflammatory diseases. In this study the pharmacokinetics and pharmacodynamics of the selective 5-lipoxygenase inhibitor, fenleuton, have been examined in the horse. Orally administered fenleuton (four 5 mg kg(-1) doses, given once daily) was absorbed from the gastrointestinal tract, and penetrated readily into tissue cage exudate, the ratio of the plasma:exudate AUC0-48h being 0.90+/-0.02 (n=6). Ionophore-stimulated leukotriene (LT) B4 synthes...
Baker JM, Stidworthy M, Gull T, Novak J, Miller JM, Antczak DF.Serological and cellular assays and molecular techniques were used to define features of the major histocompatibility complex (MHC) of the donkey With this information in hand, immune recognition of MHC determinants within and between donkeys and horses was compared. An antibody-mediated, complement-dependent, microcytotoxicity assay using a variety of antisera to donkey histocompatibility antigens, including those induced as a result of intraspecies or interspecies pregnancy in horse mares and jenny donkeys, delineated five donkey leukocyte antigen (DoLA) specificities. Antisera raised across...
Santschi EM, Grindem CB, Tate LP, Corbett WT.Ten student surgery ponies were subjected to exploratory laparotomy. Abdominal paracentesis was performed preoperatively and daily postoperatively for 6 days, then the ponies were euthanatized and necropsied. Initial baseline peritoneal fluid parameters were within established reference limits. Postoperatively, the total leukocyte count and total protein in the peritoneal fluid rose and remained elevated for the 6 days of the study. Complete blood counts (CBCs) were performed preoperatively and on days 1 and 4 postoperatively. On day 1, a stress leukogram with a mild inflammatory component dev...
Goranov Kh.The alkaline phosphatase enzyme, isolated by Morton's method from leukocytes of sheep, goats, and pigs gave after agarose elctrophoresis two isoenzyme fractions moving to the positive pole at the sites of the alpha 1- and alpha 2-globulins of the blood serum. In bovine leukocytes, besides these two fractions there was a third one that moved more slowly in the zone of the beta-globulins. In horses the alkaline phosphatase of leukocytes produced a wide band within the zones of the beta-globulins and the albumins. It was established that the proportion between the individual isoenzyme fractions o...
Auer DE.The role of oxygen-derived free radicals is considered critical to the etio-pathogenesis of equine inflammatory joint disease. In vivo, the superoxide radical in the joint may be derived either from activated polymorphonuclear leukocytes or from an ischemia/reperfusion cycle. In the presence of ferrous iron, it may generate the highly reactive hydroxyl radical (OH *). Predisposing factors may include synovitis, exercise-induced ischemia and minor traumatic injury to the joints. Unlike other inflammatory mediators, oxygen-derived free radicals may damage tissue directly and these reactive speci...
Grünig G, Barbis DP, Zhang CH, Davis WC, Lunn DP, Antczak DF.Equine peripheral blood lymphocytes (PBL) were enriched by positive selection using panning with a mixture of monoclonal antibodies against putative equine CD4 (Equine Leucocyte Antigen Workshop antibodies WS 1 and WS 72), or CD8 molecules (Workshop antibodies WS 12, WS 49, and WS 74). RNA was extracted from CD4 enriched cells (99% purity), from CD8 enriched cells (69% purity), from peripheral blood lymphocytes, and from neonatal equine thymus. RNA extracted from equine granulocytes and from equine kidney served as negative control. The RNA was electrophoresed in agarose and transferred to nyl...
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...
Hedges JF, Demaula CD, Moore BD, McLaughlin BE, Simon SI, MacLachlan NJ.Expression of E-selectin on activated endothelium is a critical initial step that leads to extravasation of leucocytes during inflammation, yet E-selectin is largely uncharacterized in several animal species including the horse. We have sequenced and compared E-selectin genes derived from activated cultures of purified equine (horse), cervid (black-tailed deer) and ovine (sheep) pulmonary artery endothelial cells (ECs). Phylogenetic and amino acid sequence comparisons indicate that bovine, cervid and ovine E-selectin are similar, whereas human and equine E-selectin are more closely related to ...
Wagner B, Radbruch A, Rohwer J, Leibold W.In this study we describe the generation of monoclonal antibodies (mAbs), which recognize different epitopes of the equine IgE constant heavy chain. Equi-murine recombinant IgE (rIgE), composed of the murine V(H)186.2 heavy chain variable region, linked to the equine IgE constant heavy chain and expressed together with the murine lambda(1) chain in J558L cells was used to immunize BALB/C mice. A total of 17 different mAbs were obtained, which recognized the rIgE heavy chain constant region. None of the mAbs reacted with monoclonal equine isotypes IgM, IgG1 (IgGa), IgG3 (IgG(T)), IgG4 (IgGb) or...
Wagner B, Siebenkotten G, Leibold W, Radbruch A.Due to characteristic clinical signs, immunoglobulins of isotype E (IgE) are believed to be involved in several allergic diseases of the horse. To date, closer investigations have been hampered by the fact that neither purified equine IgE nor anti-equine IgE monoclonal antibodies were available for IgE isotype determination. As an approach to solve this problem, we constructed a stable cell line (EqE6) that expresses recombinant equi-murine IgE specific for 4-(hydroxy-3-nitro-phenyl) acetyl (NP). Biochemical analysis of the purified protein revealed a highly glycosilated IgE monomer of approxi...
Lambertini C, Bombardi C, Zannoni A, Bernardini C, Dondi F, Morini M, Rinnovati R, Spadari A, Romagnoli N.Proteinase activated receptor 4 (PAR) in the gastrointestinal tract is involved in the regulation of inflammation and pain pathways. The aim of the present study was to evaluate the distribution and expression of PAR in the jejunum of healthy horses and in the pathologic tracts from horses undergoing surgery for herniation of the small intestine through the epiploic foramen. Eight healthy horses (Group H) and eight horses with epiploic hernia (Group EH) were included; the jejunum samples were collected at the slaughter or intraoperatively after enterectomy, respectively. To evaluate PAR expres...
Dinev D, Dzhurov A.An experiment in comparative developmental aspect was carried out with 5 horses and 5 cattle having a total of 60 skin and muscular wounds. Light microscopic studies on hematoxylin-eosin stained preparations revealed that the dystrophic changes in the underlying tissues were more strongly manifested in horses. This had to be taken into consideration in the surgical treatment of wounds with this animal species. The basic differences comprised the better manifested leukocytic reaction in horses and the more strongly expressed processes of growth and differentiation on the part of the connective ...
Köhne M, Kirch F, Tönissen A, Martinsson G, Rabe U, Sieme H, Schuberth HJ.To improve accuracy in evaluating stallion ejaculates, an antibody-based, flow cytometric assay for the detection and identification of leukocyte subpopulations (CD4-, CD8-, CD21-, CD172a-positive cells) in stallion semen (n = 12) was established. For establishment of the assay, native semen was supplemented with blood leukocytes (control: 20% leukocytes, 80% sperm cells) and analysed by flow cytometry. Adding antioxidants (ascorbic acid and butylated hydroxytoluol) to semen immediately after collection inhibited rapid death of lymphoid cells in sperm leukocyte mixtures. In control set-ups...
Degroote RL, Hauck SM, Kremmer E, Amann B, Ueffing M, Deeg CA.The molecular mechanism which enables activated immune cells to cross the blood-retinal barrier in spontaneous autoimmune uveitis is yet to be unraveled. Equine recurrent uveitis is the only spontaneous animal model allowing us to investigate the autoimmune mediated transformation of leukocytes in the course of this sight threatening disease. Hypothesizing that peripheral blood immune cells change their protein expression pattern in spontaneous autoimmune uveitis, we used DIGE to detect proteins with altered abundance comparing peripheral immune cells of healthy and ERU diseased horses. Among ...
Rapacz-Leonard A, Dąbrowska M, Janowski T.During pregnancy in larger mammals, the maternal immune system must tolerate the fetus for months while resisting external infection. This tolerance is facilitated by immunological communication between the fetus and the mother, which is mediated by Major Histocompatibility Complex I (MHC I) proteins, by leukocytes, and by the cytokines secreted by the leukocytes. Fetal-maternal immunological communication also supports pregnancy by inducing physiological changes in the mother. If the mother "misunderstands" the signal sent by the fetus during pregnancy, the fetus will be miscarried or deliver...
