White blood cells (WBCs) are integral components of the equine immune system, responsible for protecting horses against infections and foreign invaders. These cells circulate in the bloodstream and are produced in the bone marrow. The primary types of white blood cells in horses include neutrophils, lymphocytes, monocytes, eosinophils, and basophils, each with distinct functions in immune defense. Neutrophils are involved in the immediate response to infections, while lymphocytes play a role in adaptive immunity. Monocytes are involved in phagocytosis and the regulation of immune responses. Eosinophils and basophils are associated with allergic reactions and parasitic infections. Variations in white blood cell counts can indicate health conditions, making them valuable in veterinary diagnostics. This page compiles peer-reviewed research studies and scholarly articles that explore the function, regulation, and clinical relevance of white blood cells in equine health.
Salinas C, Espinosa G, Morales N, HenrÃquez C, Morán G, Gajardo G, Uberti B.Obesity is a highly prevalent condition in horses. Dysfunctional neutrophil activity has been reported in metabolically healthy obese humans, but minimal data exist regarding horses. The present study evaluated the effect of obesity on apoptosis, phagocytosis and oxidative burst activity of peripheral blood neutrophils from lean and obese non-insulin dysregulated horses. Seven lean (BCS, body condition score 4-6/9) and five obese (BCS 8-9) horses were enrolled in the study. All animals underwent two metabolic tests (OGT, oral glucose test; IRT, insulin response test) before their selection to ...
Adler DMT, Serteyn D, Franck T, Jørgensen E, Christophersen MT, Denwood M, Verwilghen DR.To compare the extent of inflammation and catabolic collagen response in the middle carpal joints (MCJs) of healthy horses following intra-articular injection of 2% lidocaine, 2% mepivacaine, lactated Ringer solution (LRS), or 0.1% methyl parahydroxybenzoate. 17 adult horses. In the first of 2 experiments, the left middle carpal joint (MCJ) of each of 12 horses was injected with 10 mL of 2% lidocaine (n = 3), 2% mepivacaine (3), or LRS (control; 6). After a 4-week washout period, the right MCJ of the horses that received lidocaine or mepivacaine was injected with 10 mL of LRS, and the right MC...
Connelly C, Norton NA, Hurley DJ, Hart KA, Meichner K, Gogal RM.Peripheral blood is commonly sampled to assess the health status of human and veterinary patients. Venous blood collection is a minimally invasive procedure, and in the horse, the common collection site is the jugular vein. Post blood collection, sample processing for leukocyte enrichment can vary by research laboratory with the potential to yield different effects on the enriched cells and their function. The focus of the present study was to compare a common blood dilution-leukocyte enrichment technique using a Histopaque gradient medium (His) to a modified leukocyte buffy coat syringe-lymph...
Sotelo EDP, Vendruscolo CP, Fülber J, Seidel SRT, Jaramillo FM, Agreste FR, Silva LCLCD, Baccarin RYA.Several studies in human and equine medicine have produced controversial results regarding the role of dimethylsulfoxide (DMSO) as a therapeutic agent. This study aimed to evaluate the effect of joint lavage with different DMSO concentrations on biomarkers of synovial fluid inflammation and cartilage degradation in joints with lipopolysaccharide (LPS)-induced synovitis. Twenty-six tibiotarsal joints of 13 horses were randomly distributed into four groups (lactated Ringer's solution; 5% DMSO in lactated Ringer's; 10% DMSO in lactated Ringer's; and sham). All animals were evaluated for the prese...
Alonso JM, Schmidt EMS, Eckersall PD, Kjelgaard-Hansen M, Alves ALG, Rodrigues CA, Watanabe MJ, Crescencio AP, Hussni CA.The acute phase response is a response to injury and depends on the severity of the trauma. Heparin is routinely used for postsurgical treatment of horses to prevent abdominal adhesions; however, its effect on inflammation is unknown. This study aimed to assess systemic inflammatory response of horses subjected to small colon enterotomy and to evaluate heparin effects on postsurgical inflammation. Ten adult horses were subjected to small colon enterotomy and were assigned to a control or a treatment group. Both groups received prophylactic antibiotics and flunixin, and the treatment group rece...
Lange-Consiglio A, Romele P, Magatti M, Silini A, Idda A, Martino NA, Cremonesi F, Parolini O.Equine amniotic mesenchymal stromal cells (AMSCs) and their conditioned medium (CM) were evaluated for their ability to inhibit in vitro proliferation of peripheral blood mononuclear cells (PBMCs) with and without priming. Additionally, AMSC immunogenicity was assessed by expression of MHCI and MHCII and their ability to counteract the in vitro inflammatory process. Horse PBMC proliferation was induced with phytohemagglutinin. AMSC priming was performed with 10 ng/ml of TNF-α, 100 ng/ml of IFN-γ, and a combination of 5 ng/ml of TNF-α and 50 ng/ml of IFN-γ. The CM generated from naÃ...
Stokol T.This article uses a case-based approach, complemented with diagnostic algorithms and images, to highlight hematologic changes of pathologic relevance in horses, namely, marked erythrocytosis, anemia or leukocytosis, inflammatory leukograms, lymphocytosis in adult horses, thrombocytopenia, and pancytopenia. These hematologic abnormalities occur with certain diseases and their identification can help clinicians narrow to down differential diagnostic lists. This article highlights the importance of blood smear examination, particularly, but not only, when numerical red flags are identified on aut...
Andrade DGA, Basso RM, Castiglioni MCR, Silva JP, Machado VMV, Laufer-Amorim R, Borges AS, Oliveira-Filho JP.Four causative mutations (D1, D2, D3*, and D4) of chondrodysplastic dwarfism have been described in the equine () gene. Homozygotes for one of these mutations and heterozygotes for any combination of these mutations exhibit the disproportionate dwarfism phenotype. However, no case description of homozygotes for D4 (D4/D4) has been reported in the literature, to our knowledge. We report 2 Miniature horses with the genotype D4/D4 in the gene. Clinically, the 2 dwarfs had a domed head that was large compared to the rest of the body, mandibular prognathism, and short and bowed limbs, mainly in t...
Fernandez NJ, Gilroy CV, Wagg CR, Kwong GPS, Roy MF.The detection of band neutrophils and toxic change via microscopic blood smear review is vitally important, as their presence indicates systemic inflammation. However, in-clinic evaluation of WBC morphology is often limited. Objective: We aimed to determine the agreement between expert raters in the detection of bands and toxic change. Methods: Three board-certified clinical pathologists each evaluated 109 blood smears from horses with acute disease, and 19 control smears from healthy horses. The pathologists determined if bands were present, and if so, the percentage of bands present. They a...
Neuenschwander HM, Moreira JJ, Vendruscolo CP, Fülber J, Seidel SRT, Michelacci YM, Baccarin RYA.The intra-articular use of hyaluronic acid (HA) for the treatment of synovitis and osteoarthritis is still controversial. As a consequence, corticosteroids remain the most frequently employed therapeutic agents, despite their potential systemic and local deleterious effects. This study examined the anti-inflammatory, antioxidant, and chondroprotective activities of low and high molecular weight hyaluronic acid (LMW-HA and HMW-HA) on lipopolysaccharide (LPS)-induced synovitis in horses compared to triamcinolone acetonide (TA). LPS was injected in the metacarpophalangeal joints, which were treat...
Espinosa G, Plaza A, Schenffeldt A, Alarcón P, Gajardo G, Uberti B, Morán G, HenrÃquez C.Polymorphonuclear neutrophils (PMN) are the largest leukocyte population in the blood of most mammals including horses, and play an important defensive role in many infectious diseases. However, the mechanisms that increase PMN as one of the main cellular subsets in the defense against pathogens could also be involved in the pathophysiology of dysregulated inflammatory conditions. Mesenchymal stem/stromal cells (MSCs) are a heterogeneous population with a modulatory potential on the inflammatory response and are known to interact with nearly all cells of the immune system, including PMN. In th...
MarÃn-López A, Barreiro-Piñeiro N, Utrilla-Trigo S, Barriales D, Benavente J, Nogales A, MartÃnez-Costas J, Ortego J, Calvo-Pinilla E.African horse sickness virus (AHSV) is an insect-borne pathogen that causes acute disease in horses and other equids. In an effort to improve the safety of currently available vaccines and to acquire new knowledge about the determinants of AHSV immunogenicity, new generation vaccines are being developed. In this work we have generated and tested a novel immunization approach comprised of nonstructural protein 1 (NS1) of AHSV serotype 4 (AHSV-4) incorporated into avian reovirus muNS protein microspheres (MS-NS1) and/or expressed using recombinant modified vaccinia virus Ankara vector (MVA-NS1)....
Wernery U, Rodriguez Caveney M, Wernery R, Raghavan R, Laroucau K, Syriac G, Thomas SM, John J, Joseph M, Jose S, Joseph S, Woo P.Six horses were challenged experimentally with a strain of Burkholderia pseudomallei isolated from a fatal case of the infection in a dromedary camel years earlier in the Emirate of Dubai. Three horses were inoculated subcutaneously and in 3 the bacterium was administered by the oral route. Four of the horses became serologically positive based on reactions to one or more of the OIE described tests for glanders. B. pseudomallei was re-isolated from the 4 serological positive horses. Only one of the subcutaneously infected horses, developed fever for 3 days. The white blood cell values and the ...
Daden R, Zarhouni FZ, Chakir J, Piro M, Achaâban MR, Ouassat M, El Allali K.To evaluate the effect of plasmapheresis on clinical, hematological, and biochemical parameters after exercise, a plasmapheresis session was realized on six jumping horses (plasmapheresis group) that underwent three consecutive days of physical graded exercise. The control group (n = 6) went through the same exercise but not subjected to the plasmapheresis session. Seventeen milliliters of plasma/kg of body weight was harvested from each horse. The procedure was well tolerated by the horses. The plasmapheresis leads to a significant increase of the hemoglobin, hematocrit, red blood cell, whit...
Carossino M, Dini P, Kalbfleisch TS, Loynachan AT, Canisso IF, Cook RF, Timoney PJ, Balasuriya UBR.Equine arteritis virus (EAV) has the unique ability to establish long-term persistent infection in the reproductive tract of stallions and be sexually transmitted. Previous studies showed that long-term persistent infection is associated with a specific allele of the CXCL16 gene (CXCL16S) and that persistence is maintained despite the presence of local inflammatory and humoral and mucosal antibody responses. Here, we performed transcriptomic analysis of the ampullae, the primary site of EAV persistence in long-term EAV carrier stallions, to understand the molecular signatures of viral persiste...
Sheta E, Farghali H, Ragab S, Hassan N, El-Sherif A.Twenty-five Thoroughbred jumper geldings suffered back soreness with poor performance, and 5 control horses were assessed by archived computer data, clinical examination, and laboratory analyses of complete blood picture, serum enzymes, and cortisol level, before and after cloprostenol-pharmacopuncture. The 25 diseased horses before therapy showed significant increases in aspartate aminotransferase and creatine phosphokinase with clinical pains scored mild in 15 horses, moderate in 9 horses, and severe in one horse, without changes in the hormonal and hematological data. After therapy, they re...
Samimi AS, Samimi K, Karimiafshar M, Tajik J.The objective of this study was to comparatively evaluate the anti-inflammatory effects of intravenous administration of insulin (in different doses) and flunixin on physiological variables, acute-phase responses, and cardiovascular biomarkers during inflammatory phase which was induced by Escherichia coli lipopolysaccharide (LPS) serotype O55:B5 in miniature donkeys. A total of twenty-four clinically healthy male adult miniature donkeys aged 5 ± 1 year and weighing 120 ± 10 kg were studied. The animals were assigned randomly to four experimental groups (three treatment and one positive co...
Witkowska-Piłaszewicz O, Bąska P, Czopowicz M, Żmigrodzka M, Szczepaniak J, Szarska E, Winnicka A, Cywińska A.Sport training leads to adaptation to physical effort that is reflected by the changes in blood parameters. In equine endurance athletes, blood testing is accepted as a support in training, however, only the changes before versus after exercise in creatine phosphokinase activity (CPK) and basic blood parameters are usually measured. This study is the first longitudinal investigation of the changes in routinely measured blood parameters and, additionally, serum amyloid A (SAA), during seven months, in Arabian horses introduced to endurance training and competing in events for young horses. It h...
Joonè CJ, Nolan MB, Bertschinger HJ, Schulman ML.A sequence of studies is reviewed that reported the domestic horse (Equus caballus) mare as an appropriate and accessible research platform for recording clinical and laboratory data post-immunisation with anti- GnRH and -zona pellucida (ZP) immunocontraceptive vaccines. Experience with a native porcine ZP (pZP) vaccine in African elephant (Loxodonta africana) cows highlighted needs for improving vaccine formulations and more clearly defining associated ovarian effects and safety profiles. Initially, the efficacy, reversibility and safety of the GnRH vaccine Improvac® in mares was demonstrate...
Bohlin A, Saegerman C, Hoeberg E, Sånge A, Nostell K, Durie I, Husted L, Öhman A, van Galen G.It is highly desirable to assess the probability of survival in sick neonatal foals upon admission. The foal survival score (FSS) is a published scoring system used to estimate the probability of survival in hospitalized neonatal foals <4 days old. Objective: To evaluate the ability of the FSS to predict survival in older foals from a geographically different area compared to the original study. Methods: Five-hundred ninety hospitalized neonatal foals ≤14 days of age. Methods: Retrospective Danish-Swedish multicenter study that included details of signalment, history, clinical examinatio...
Faramarzi B, Rich L.Foals' haematological values change constantly during their first year of life. The use of updated age-based reference intervals (RIs) is imperative for providing accurate diagnosis and optimum care for sick foals. The authors' objective was to provide updated RIs for 13 haematological values in 2, 7, 14, 30, 90, 180 and 365-day-old foals and to investigate the changes over time in each measured value. Venous blood was collected at those ages from clinically healthy foals. Thirteen haematological values were analysed. The 95% RIs were reported using a bootstrapping method. Differences over tim...
Soroko M, Śpitalniak-Bajerska K, Zaborski D, Poźniak B, Dudek K, Janczarek I.The aim of the study was to assess the effects of training on haematological and biochemical blood parameters as well as on the changes in body surface temperature in horses. In order to identify the predictive value of surface temperature measurements as a marker of animal's performance, their correlations with blood parameters were investigated. The study was carried out on nine horses divided into two groups: routinely ridden and never ridden. Infrared thermography was used to assess surface temperature changes before (BT) and just after training (JAT) on a treadmill. Seven regions of int...
Witonsky S, Buechner-Maxwell V, Santonastasto A, Pleasant R, Werre S, Wagner B, Ellison S, Lindsay D.Equine protozoal myeloencephalitis (EPM) is a common and devastating neurologic disease of horses in the United States. Because some EPM-affected horses have decreased immune responses, immunomodulators such as levamisole have been proposed as supplemental treatments. However, little is known about levamisole's effects or its mechanism of action in horses. Objective: Levamisole in combination with another mitogen will stimulate a macrophage 1 (M1), dendritic cell 1 (DC1), T-helper 1 (CD4 Th1), and T-cytotoxic (CD8) immune response in equine peripheral blood mononuclear cells (PBMCs) in vitro a...
Henkel J, Lafayette C, Brooks SA, Martin K, Patterson-Rosa L, Cook D, Jagannathan V, Leeb T.White spotting phenotypes in horses are highly valued in some breeds. They are quite variable and may range from the common white markings up to completely white horses. EDNRB, KIT, MITF, PAX3 and TRPM1 represent known candidate genes for white spotting phenotypes in horses. For the present study, we investigated an American Paint Horse family segregating a phenotype involving white spotting and blue eyes. Six of eight horses with the white-spotting phenotype were deaf. We obtained whole-genome sequence data from an affected horse and specifically searched for structural variants in the known ...
Silvestre-Ferreira AC, Cotovio M, Maia M, Queiroga F, Pires MJ, Colaço A.The Lusitano horse is an autochthonous Portuguese breed with a growing worldwide expansion. Our objective was to establish reference intervals for haematological parameters using the haematological cell counter LaserCyte (IDEXX). For this purpose, blood samples from 100 healthy adult horses (13 females and 87 males, ranging from 3 to 25 years of age) were analysed. The reference intervals were estimated following the ASVCP guidelines with the Reference Value Advisor software. The obtained reference intervals were 6.4-10.1 × 1012/L for red blood cells, 30.6-45.1% for haematocrit, 11.6-17.1 g/d...
Roszkowska K, Witkowska-Pilaszewicz O, Przewozny M, Cywinska A.Whole body and partial body cryotherapies (WBC and PBC) have been successfully used in human medicine, and currently also are being proposed in veterinary practice.In horses, only the partial body cryotherapy provided in cryosauna is considered, due to the technical conditions. These therapies have been dedicated to human patients with rheumatic and inflammatory diseases as well as an assistance during training in athletes. The anti-inflammatory effects have been demonstrated clinically and indicated by the changes in several hematological and immunological parameters, however, various pattern...
Joonè CJ, Schulman ML, Fosgate GT, Plagis TA, Crafford JE, Gupta SK, Bertschinger HJ.Few studies have investigated the cell mediated immune response during zona pellucida-based immunocontraception, despite hypothesized cytotoxic T-cell involvement in ovarian dysfunction associated with these vaccines. This study aimed to investigate antigen-specific anamnestic responses of helper (CD4) and cytotoxic (CD8) T-lymphocytes in peripheral blood mononuclear cells (PBMC) isolated from pony mares before and after their treatment with native porcine zona pellucida (pZP), recombinant pZP3 and pZP4 antigens (reZP) or adjuvanted saline. Mares were randomly assigned to pZP, reZP and control...
Wimer CL, Schnabel CL, Perkins G, Babasyan S, Freer H, Stout AE, Rollins A, Osterrieder N, Goodman LB, Glaser A, Wagner B.The equine herpesvirus type 1 (EHV-1) ORF1 and ORF71 genes have immune modulatory effects in vitro. Experimental infection of horses using virus mutants with multiple deletions including ORF1 and ORF71 showed promise as vaccine candidates against EHV-1. Here, the combined effects of ORF1 and ORF71 deletions from the neuropathogenic EHV-1 strain Ab4 on clinical disease and host immune response were further explored. Three groups of EHV-1 naïve horses were experimentally infected with the ORF1/71 gene deletion mutant (Ab4ΔORF1/71), the parent Ab4 strain, or remained uninfected. In comparison t...
Hellman S, Hjertner B, Morein B, Fossum C.The immunomodulatory effect of a new particulate adjuvant, G3, alone or in combination with agonists to TLR2/1 or TLR5 was evaluated in cultures of equine PBMC. Exposure to the G3 adjuvant up-regulated genes encoding IFN-γ, IL-1β, IL-6, IL-8, IL-12p40 and IL-23p19 in the majority of the horses tested, indicating that the G3 adjuvant induced a pro-inflammatory and Th1 dominated profile. In accordance, genes encoding IL-13, IL-4, IL-10 and TGF-β remained unaffected and genes encoding IFN-α, IL-17A and TNF-α were only occasionally and weakly induced. The two TLR agonists Pam3CSK4 (TLR2/1) an...
Chikhaoui M, Smail F, Adda F.The Barb horse this famous Maghreb horse, symbol of the pride of the Berber peoples It is particularly significant in the Algerian history, tradition and culture. This study consists in establishing the hematologic values specific to the Barb horse in the national stud farm "Chaou-Chaoua" in Tiaret. Algeria. Age of the study animals varied between 1 and 23 years. The effects of age and sex were analyzed by dividing animals into four groups, A (1 - 2 years, n=5), B (3-5 years, n=5), C (5-10 years, n=5) and D (11-23 years, n=5); for females and E (2 - 3 years, n=5), F (5-7, n=5), G (10, n=5) and...
Sutter WW, Kaneps AJ, Bertone AL.To evaluate the buffy coat and apheresis methods for preparation of platelet concentrates from equine blood by comparing platelet and growth factor concentrations. Methods: 15 mature mixed-breed geldings. Methods: Whole blood samples were collected and processed by use of a buffy coat or apheresis method to obtain platelet poor and platelet concentrated fractions. The PCV, WBC count, and platelet count were compared among whole blood samples, platelet poor fractions, concentrates obtained by use of the apheresis method (ie, apheresis platelet concentrates), and concentrates obtained by use of ...
Kleiber C, McGorum BC, Horohov DW, Pirie RS, Zurbriggen A, Straub R.Equine recurrent airway obstruction (RAO) is thought to result from an aberrant immune response to inhaled antigens, modulated by T lymphocytes via the secretion of pro-inflammatory cytokines. However data relating to the phenotypes of the T lymphocytes present in peripheral blood and bronchoalveolar lavage fluid of RAO horses and their cytokine profiles are contradictory. The aim of this study was to further investigate the cytokine (IL-4, IL-5, IL-13 and INF-gamma) mRNA expression profile in peripheral blood lymphocytes and bronchoalveolar lavage lymphocytes from RAO and control horses, befo...
Canisso IF, Ball BA, Cray C, Williams NM, Scoggin KE, Davolli GM, Squires EL, Troedsson MH.Currently, placentitis, an important cause of late pregnancy loss in mares, is diagnosed by clinical signs and ultrasonography. Acute phase proteins (APP) are mainly produced and secreted by the liver in response to acute inflammatory stimuli. We hypothesized that APP are increased in mares with placentitis. Methods: Concentrations of serum amyloid A (SAA), haptoglobin (Hp), fibrinogen (Fb), and white blood cell counts (WBC) were determined in plasma of mares with experimentally induced placentitis and gestationally age-matched control mares. Placentitis was induced via intracervical inoculati...
Capomaccio S, Cappelli K, Barrey E, Felicetti M, Silvestrelli M, Verini-Supplizi A.It is known that moderate physical activity may have beneficial effects on health, whereas strenuous effort induces a state resembling inflammation. The molecular mechanisms underlying the cellular response to exercise remain unclear, although it is clear that the immune system plays a key role. It has been hypothesized that the physio-pathological condition that develops in athletes subjected to heavy training is caused by derangement of cellular immune regulation. The purpose of the present study was to obtain information on endurance horse gene transcription under strenuous conditions and t...
Tumas DB, Hines MT, Perryman LE, Davis WC, McGuire TC.The immune control of chronic equine infectious anaemia (EIA) lentiviral infection was investigated by specifically depleting CD5+ T lymphocytes in vivo with monoclonal antibody (MAb) or by immunosuppression with corticosteroids. MAb was given at 25 to 50 mg/day intravenously for 11 days. Murine IgG1 anti-equine CD2 MAb (n = 2 horses) or IgG1 (n = 2) and IgG2a control MAb (n = 2 normal; 2 EIA-infected) did not deplete CD2+ T lymphocytes in horses. Horses given murine IgG2a anti-CD5 MAb HB19A (n = 4 normal; 5 EIA-infected) had depletion of peripheral blood CD5+ T lymphocytes during treatment. T...
Hammond SA, Cook SJ, Falo LD, Issel CJ, Montelaro RC.Immunization regimens that induce a broadly reactive cytolytic T lymphocyte (CTL) response specific for lentiviral antigens have emerged as the leading candidates in efficacy trials conducted in both animal modelshumans. To date, lentivirus vaccination strategies have overlooked one such immunization strategy, namely the use of particulate antigens. To evaluate the efficacy of targeting antigen into the phagocytic pathway to elicit a cell-mediated immune response to lentiviral antigens, we initiated the first study of a particulate-based vaccination protocol using a large animal model system. ...
Derksen FJ, Scott JS, Miller DC, Slocombe RF, Robinson NE.We performed bronchoalveolar lavage (BAL) in 2 groups of ponies. Principal ponies had a history of heaves, a disease characterized by recurrent airway obstruction and airway hyperreactivity when ponies are housed in a barn and fed hay; control ponies had no history of airway obstruction. Ponies were paired (principal and control), and BAL was performed after 2 months of being pastured when principal ponies were in clinical remission (Period A), after barn housing when principal ponies had acute airway obstruction (Period B), and after a 1- and a 2-wk recovery phase of pasture grazing (Periods ...
Mealey RH, Sharif A, Ellis SA, Littke MH, Leib SR, McGuire TC.Cytotoxic T lymphocytes (CTL) are critical for control of lentiviruses, including equine infectious anemia virus (EIAV). Measurement of equine CTL responses has relied on chromium-release assays, which do not allow accurate quantitation. Recently, the equine MHC class I molecule 7-6, associated with the ELA-A1 haplotype, was shown to present both the Gag-GW12 and Env-RW12 EIAV CTL epitopes. In this study, 7-6/Gag-GW12 and 7-6/Env-RW12 MHC class I/peptide tetrameric complexes were constructed and used to analyze Gag-GW12- and Env-RW12-specific CTL responses in two EIAV-infected horses (A2164 an...
Ibrahim S, Saunders K, Kydd JH, Lunn DP, Steinbach F.Three hundred and seventy-nine monoclonal antibodies (mAbs) against various human CD molecules supplied to the HLDA8 animal homologues section (including four isotype controls) were analysed for cross-reactivity with equine leukocytes. First, flow cytometric identification of positively reacting mAbs was performed in one laboratory. Thereafter, a second round of flow cytometric evaluation was performed, involving three laboratories participating in the study. The first test-round indicated 17 mAbs as potentially positive. After the second round of flow cytometric analysis, 14 mAbs remained (di...
Robbin MG, Wagner B, Noronha LE, Antczak DF, de Mestre AM.Several distinct T lymphocyte subpopulations with immunoregulatory activity have been described in a number of mammalian species. This study performed a phenotypic analysis of cells expressing regulatory T cell (Treg) markers in the peripheral blood of a cohort of 18 horses aged 6 months to 23 years, using antibodies to both intracellular and cell surface markers, including Forkhead box P3 (FOXP3), CD4, CD8, CD25, interferon gamma (IFNγ) and interleukin 10 (IL-10). In peripheral blood, a mean of 2.2 ± 0.2% CD4+ and 0.5 ± 0.1% CD8+ lymphocytes expressed FOXP3. The mean percentage of CD4+FOXP...
Castagnetti C, Mariella J, Pirrone A, Cinotti S, Mari G, Peli A.To evaluate and compare the gene expression of interleukin(IL)-1β, IL-8, and interferon-γ during the first 72 hours after birth in healthy foals and during the first 72 hours after hospitalization in sick neonatal foals and investigate correlations of clinicopathologic variables with cytokine expressions in healthy and sick neonatal foals. Methods: 33 foals < 7 days old (10 healthy foals, 7 foals with sepsis, 6 foals with peripartum asphyxia syndrome, and 12 foals with other diseases [2 with failure of passive transfer of immunity only were not further evaluated]). Methods: A blood sample...
McFarlane D, Sellon DC, Gibbs SA.To characterize age-associated changes in lymphocyte population subsets and immunoglobulin isotypes. Methods: 30 healthy young light-breed horses (5 to 12 years old) and 30 healthy aged light-breed horses (> 20 years old). Methods: Lymphocyte subset populations were identified, using monoclonal antibodies to cell surface markers CD5, CD4, CD8, and IgG. Subset populations were quantitated by use of flow cytometric analysis of antibody-stained cells. Serum immunoglobulin concentration was determined using single radial immunodiffusion. Results: Absolute cell counts of total lymphocytes, T cells,...
Carossino M, Loynachan AT, Canisso IF, Cook RF, Campos JR, Nam B, Go YY, Squires EL, Troedsson MHT, Swerczek T, Del Piero F, Bailey E, Timoney PJ....Equine arteritis virus (EAV) has a global impact on the equine industry as the causative agent of equine viral arteritis (EVA), a respiratory, systemic, and reproductive disease of equids. A distinctive feature of EAV infection is that it establishes long-term persistent infection in 10 to 70% of infected stallions (carriers). In these stallions, EAV is detectable only in the reproductive tract, and viral persistence occurs despite the presence of high serum neutralizing antibody titers. Carrier stallions constitute the natural reservoir of the virus as they continuously shed EAV in their seme...
Moraes AP, Moreira JJ, Brossi PM, Machado TS, Michelacci YM, Baccarin RY.This study aimed to verify whether transient inflammatory reactions incited by the administration of intra-articular platelet-rich plasma (PRP) affected joint components through short- and long-term in vivo evaluation of inflammatory biomarkers and extracellular matrix degradation products in synovial fluid. The effects of PRP were analyzed in a short phase protocol (SPP) and in a prolonged phase protocol (PPP), using saline-injected joints as controls. In the SPP, higher white blood cell counts and prostaglandin E2 and total protein concentrations were observed in the synovial fluid of PRP-tr...
Dawson TRMY, Horohov DW, Meijer WG, Muscatello G.Rhodococcus equi is recognised to cause chronic purulent bronchopneumonia in foals of less than 6 months of age. Virulent strains of the bacteria possess a large 80-90 kb plasmid encoding several virulence-associated proteins, including virulence-associated protein A (VapA), which is associated with disease. R. equi pneumonia can represent significant costs and wastage to the equine breeding industry, especially on stud farms where the disease is endemic. This article reviews knowledge of the equine immune response, both in the immune adult and susceptible neonate, with respect to this pathoge...
Moore BR, Krakowka S, Robertson JT, Cummins JM.Cytologic examination of bronchoalveolar lavage fluid (BALF), including phenotypic analysis of lymphocytes, was performed on 32 Standardbreds with poor race performance and endoscopic examination findings characteristic of inflammatory airway disease (IAD). Nucleated cell counts in BALF from IAD-affected horses were higher than those in control horses; the cytologic profile of BALF in affected horses included mixed inflammation, characterized by mild neutrophilia, lymphocytosis, and monocytosis. Eosinophil and mast cell counts were not higher in the IAD-affected group, compared with those in t...
Sheats MK, Pescosolido KC, Hefner EM, Sung EJ, Adler KB, Jones SL.Neutrophil infiltration is a prominent feature in a number of pathologic conditions affecting horses including recurrent airway obstruction, ischemia-reperfusion injury, and laminitis. Cell signaling components involved in neutrophil migration represent targets for novel anti-inflammatory therapies. In order to migrate into tissue, neutrophils must respond to chemoattractant signals in their external environment through activation of adhesion receptors (i.e. integrins) and reorganization of the actin cytoskeleton. Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS), a highly conserved actin...
Faubladier C, Chaucheyras-Durand F, da Veiga L, Julliand V.This study evaluated the effect of transportation on fecal bacterial communities and activities in horses with or without supplementation of live yeast and attempted to link those effects with changes in blood stress markers. Four mature horses were assigned to a crossover design and fed a basal diet (60:40 forage to concentrate; 1.45% BW on a DM basis), with or without supplementation, of 2 × 10(10) cfu/d of Saccharomyces cerevisiae CNCM I-1077. After a 14-d adaptation to dietary treatments, the 5-d experiment started 1 d before transportation (d -1). At d 0, horses were simultaneously trans...
Flaminio MJ, Antczak DF.At days 36-38 of gestation, the equine invasive trophoblast cells migrate into the endometrium of the pregnant mare to form the endometrial cups. During their migration, they become surrounded by maternal CD4+ and CD8+ T lymphocytes, and stimulate a cytotoxic antibody response to the paternal major histocompatibility complex class I antigens that they express. Nevertheless, endometrial cup cells remain viable at the site of uterine invasion up to days 80-100 of gestation, suggesting the participation of immunomodulatory mechanisms to the maternal cellular immune response. To determine the effe...
Tonkin DR, Whitmore A, Johnston RE, Barro M.Replicon particles derived from Venezuelan equine encephalitis virus (VEE) are infectious non-propagating particles which act as a safe and potent systemic, mucosal, and cellular adjuvant when delivered with antigen. VEE and VEE replicon particles (VRP) can target multiple cell types including dendritic cells (DCs). The role of these cell types in VRP adjuvant activity has not been previously evaluated, and for these studies we focused on the contribution of DCs to the response to VRP. By analysis of VRP targeting in the draining lymph node, we found that VRP induced rapid recruitment of TNF-s...
Trapp S, von Einem J, Hofmann H, Köstler J, Wild J, Wagner R, Beer M, Osterrieder N.Key problems using viral vectors for vaccination and gene therapy are antivector immunity, low transduction efficiencies, acute toxicity, and limited capacity to package foreign genetic information. It could be demonstrated that animal and human cells were efficiently transduced with equine herpesvirus 1 (EHV-1) reconstituted from viral DNA maintained and manipulated in Escherichia coli. Between 13 and 23% of primary human CD3+, CD4+, CD8+, CD11b+, and CD19+ cells and more than 70% of CD4+ MT4 cells or various human tumor cell lines (MeWo, Huh7, HeLa, 293T, or H1299) could be transduced with o...
Giguère S, Hernandez J, Gaskin J, Miller C, Bowman JL.To evaluate WBC concentration, plasma fibrinogen concentration, and an agar gel immunodiffusion (AGID) test for early identification of Rhodococcus equi-infected foals. Methods: Prospective study. Methods: 162 foals from a farm with enzootic R equi infection. Methods: Blood samples were obtained from each foal at 4-week intervals for measurement of WBC and plasma fibrinogen concentrations and at 2-week intervals for detection of anti-R equi antibody by an AGID assay. Diagnostic performance of WBC and fibrinogen concentrations was assessed by use of receiver operating characteristic curve analy...
Andreassen SM, Vinther AML, Nielsen SS, Andersen PH, Tnibar A, Kristensen AT, Jacobsen S.Septic arthritis is a common and potentially devastating disease characterized by severe intra-articular (IA) inflammation and fibrin deposition. Research into equine joint pathologies has focused on inflammation, but recent research in humans suggests that both haemostatic and inflammatory pathways are activated in the joint compartment in arthritic conditions. The aim of this study was to characterize the IA haemostatic and inflammatory responses in horses with experimental lipopolysaccharide (LPS)-induced joint inflammation. Inflammation was induced by IA injection of LPS into one antebrach...
Little D, Tomlinson JE, Blikslager AT.Post operative ileus (POI) remains an important cause of post operative morbidity and mortality in the horse. However, clinical progression of naturally occurring cases of POI in both horse and man does not entirely support the 'neurogenic' hypothesis as the sole mechanism of POI; and the hypothesis that inflammation plays a major role at 12-24 h after surgery requires validation. Objective: An inflammatory infiltrate in the muscularis externa and myenteric plexus of equine jejunum is present 18 h following a period of ischaemia. Methods: Samples of normal jejunum, jejunum from the proximal re...
Schnabel CL, Wemette M, Babasyan S, Freer H, Baldwin C, Wagner B.Chemokines are soluble molecules directing immune cell trafficking and homing, mediating inflammation, and initiating immune responses to infection. In horses, the analysis of chemokines has been limited by the lack of specific antibodies. We generated mAbs specific for the equine C-C motif chemokine ligands (CCL) CCL2 (MCP-1), CCL3 (MIP-1α), CCL5 (RANTES) and CCL11 (eotaxin) using hybridoma technology. Antibody specificity was confirmed by intracellular staining of Chinese Hamster Ovary cells transfected with expression vectors encoding for CCL2, CCL3, CCL5, or CCL11. Transfectants were stai...
Naskou MC, Norton NA, Copland IB, Galipeau J, Peroni JF.Platelet lysate (PL) has been extensively used for the laboratory expansion of human mesenchymal stem cells (MSC) in order to avoid fetal bovine serum (FBS) which has been associated with immune-mediated host reactions and transmission of bovine-origin microbial contaminants. Before suggesting the routine use of PL for MSC culture, we wanted to further investigate whether PL alone might trigger inflammatory responses when exposed to reactive white blood cells such as monocytes. Our objectives were to evaluate the inflammatory profile of equine monocytes cultured with equine PL (ePL) and to det...
Morris DD, Crowe N, Moore JN.Serum tumor necrosis factor (TNF) activity was quantitated in 8 horses given an IV infusion of endotoxin (0.03 micrograms of lipopolysaccharide/kg of body weight, from Escherichia coli 055:B5) in 0.9% NaCl solution over 1 hour. Serum TNF activity was likewise measured in 6 horses given only 0.9% sterile NaCl solution at the same rate. The duration of serum TNF activity was determined, and serum TNF activity was correlated with clinical and laboratory changes during the induced endotoxemia. Horses had no serum TNF activity prior to endotoxin administration, but geometric mean serum TNF activity...
Wilsterman S, Soboll-Hussey G, Lunn DP, Ashton LV, Callan RJ, Hussey SB, Rao S, Goehring LS.Infection with equine herpesvirus-1 (EHV-1) causes respiratory disease, late term abortions and equine herpesvirus myeloencephalitis (EHM) and remains an important problem in horses worldwide. Despite increasing outbreaks of EHM in recent years, our understanding of EHM pathogenesis is still limited except for the knowledge that a cell-associated viremia in peripheral blood mononuclear cells (PBMCs) is a critical link between primary respiratory EHV-1 infection and secondary complications such as late-term abortion or EHM. To address this question our objective was to identify which PBMC subpo...
Fontenot RL, Sink CA, Werre SR, Weinstein NM, Dahlgren LA.This study evaluated the quality and bacteriologic safety of platelet-rich plasma (PRP) produced by 3 simple, inexpensive tube centrifugation methods and a commercial system. Citrated equine blood collected from 26 normal horses was processed by 4 methods: blood collection tubes centrifuged at 1200 and 2000 × g, 50-mL conical tube, and a commercial system. White blood cell (WBC), red blood cell (RBC), and platelet counts and mean platelet volume (MPV) were determined for whole blood and PRP, and aerobic and anaerobic cultures were performed. Mean platelet concentrations ranged from 1.55- to 2...
Kim HY, Mott J, Zhi N, Tajima T, Rikihisa Y.Human granulocytic ehrlichiosis (HGE), a tick-borne zoonosis, is caused by an obligatory intragranulocytic bacterium, the HGE agent, a strain of Anaplasma phagocytophila. The equine model of HGE is considered valuable in understanding pathogenic and immune mechanisms of HGE. In the present study, cytokine mRNA expression by peripheral blood leukocytes (PBLs) in horses was examined during the course of infection by intravenous inoculation of A. phagocytophila or by allowing feeding by infected ticks. The p44 genes encoding the major outer membrane protein P44s of A. phagocytophila were detected...
