The equine immune system is a complex network of cells, tissues, and organs that work collaboratively to defend against pathogens and maintain homeostasis. It consists of innate and adaptive components, each with distinct functions and mechanisms. The innate immune system provides the first line of defense through physical barriers, phagocytic cells, and the complement system. The adaptive immune system involves lymphocytes, such as B cells and T cells, which generate specific responses to antigens and provide immunological memory. Research in equine immunology explores the interactions between these components, the impact of genetic and environmental factors on immune function, and the development of vaccines and therapeutics. This page gathers peer-reviewed studies and scholarly articles focusing on the mechanisms, regulation, and clinical applications of the equine immune system in health and disease.
Theoret CL, Barber SM, Gordon JR.To describe the localization of immunoreactive transforming growth factor (TGF)-beta1 in both normal skin and full-thickness dermal wounds of the limb and the thorax of the horse. Methods: Six full-thickness excisional wounds were created on the lateral aspect of one metacarpal region and on the midthoracic area of each horse. Sequentially collected tissue specimens from wound margins were assessed for TGF-beta1 expression by immunohistochemistry. Methods: Four horses (2 to 4 years of age). Methods: A neutralizing monoclonal anti-human TGF-beta1 antibody was used to detect the spatial expressi...
Jeronimo C, Archambault D.Equine arteritis virus (EAV), an enveloped positive-stranded RNA virus, is the prototype of the arterivirus group. In a previous paper (A. Kheyar, S. Martin, G. St.-Laurent, P. J. Timoney, W. H. McCollum, and D. Archambault, Clin. Diagn. Lab. Immunol. 4:648-652, 1997), we have shown that the unglycosylated membrane (M) protein, which is composed of 162 amino acids (aa), is a major target of equine antibody to EAV. In order to determine the antigenic regions of the M protein, the cDNA encoding the M protein of EAV was inserted into the procaryotic expression vector pGEX-4T-1 to produce recombin...
Learmonth GS, Love DN, Wellington JE, Gilkerson JR, Whalley JM.The unusual mucin-like high molecular mass (Mr) glycoprotein 2 (gp2) has only been described in the equid alphaherpesviruses, among which there is considerable antigenic cross-reactivity. Equine herpesvirus 1 (EHV-1) gp2 is cleaved into a highly glycosylated N-terminal subunit and a 42 kDa C-terminal cleavage product. In order to investigate their antigenic recognition by horses naturally infected with EHV-1 and/or equine herpesvirus 4 (EHV-4), the C-terminal cleavage product and high Mr gp2 were affinity purified. Cross-reactivity between EHV-1 and EHV-4 was observed for the high Mr gp2 using...
Iqbal J, Purewal AS, Edington N.The aim of this study was to investigate the role of immediate early gene (gene63) in the pathogenesis of equine herpesvirus 1 (EHV-1) acute and latent infections in equine and murine models. EHV-1 gene63 mutant virus (g63mut) along with EHV-1 (Ab4) was used for intracerebral and intranasal infection of 3 and 17-day-old mice. Both viruses were recovered at the same frequency from tissues after infection. Two Welsh ponies were infected via the intranasal route with each of the viruses. Acute infection was monitored by virus isolation from nasal swabs and peripheral blood leukocytes. Six weeks p...
Smith R, Chaffin MK, Cohen ND, Martens RJ.To characterize changes in lymphocyte subsets over time in foals from birth to 18 weeks of age, accounting for differences among individuals, and to determine the effect of overnight storage of blood samples on foal lymphocyte subset concentrations. Methods: 8 healthy Quarter Horse foals from birth to 18 weeks of age. Methods: Blood samples were collected longitudinally from birth to 18 weeks of age and a CBC performed on each sample. The samples were stained for lymphocyte markers, either immediately or after overnight storage and analyzed by flow cytometry. Results: Total leukocytes, total l...
Kingston JK, Bayly WM, Sellon DC, Meyers KM, Wardrop KJ.To investigate the potential use of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-human thrombospondin antibody for detection of the activation of equine platelets by use of flow cytometry. Methods: Platelets obtained from 6 Thoroughbreds. Methods: Flow cytometry was used to assess platelet activation as indicated by detection of binding of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-thrombospondin antibody to unactivated and ADP-, collagen-, platelet activating factor (PAF)-, and A23187-activated equine platelets. Human platelets were used...
Harrington DJ, Sutcliffe IC, Chanter N.Streptococcus equi is the aetiological agent of strangles, one of the most prevalent diseases of the horse. The animal suffering and economic burden associated with this disease necessitate effective treatment. Current antibiotic therapy is often ineffective and thus recent attention has focused on vaccine development. A systematic understanding of S. equi virulence, leading to the identification of targets to which protective immunity can be directed, is a prerequisite of the development of such a vaccine. Here, the virulence factors of S. equi are reviewed.
Palombo EA.Rotaviruses are the major cause of severe gastroenteritis in young children and animals. The rotavirus genome is composed of eleven segments of double-stranded RNA and can undergo genetic reassortment during mixed infections, leading to progeny viruses with novel or atypical phenotypes. There are numerous descriptions of rotavirus strains isolated from human and animals that share genetic and antigenic features of viruses from heterologous species. In many cases, genetic analysis by hybridization has clearly demonstrated the genetic relatedness of gene segments to those from viruses isolated f...
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,...
Fraser DG, Oaks JL, Brown WC, McGuire TC.Equine infectious anaemia virus (EIAV) is a horse lentivirus causing lifelong, persistent infection. During acute infection, CD8(+) cytotoxic T lymphocytes (CTL) are probably involved in terminating plasma viraemia. However, only a few EIAV CTL epitopes, restricted to fewer horse major histocompatibility complex (MHC) class I alleles, are known. As interferon-gamma (IFN-gamma)-secreting CD4(+), T helper 1 (Th1) lymphocytes promote CTL activity and help maintain memory CTL, identifying broadly recognized EIAV Th1 epitopes would contribute significantly to vaccine strategies seeking to promote s...
Ridgely SL, McGuire TC.The immunogenicity of equine infectious anemia virus (EIAV) Gag and Env equine leukocyte alloantigen (ELA)-A5.1, -A9, and -A1 restricted cytotoxic T lymphocyte (CTL) epitopes synthesized on multiple antigenic peptide (MAP) system coupled to tripalmitoyl-S-glycerylcysteine (P3C) was evaluated in vitro. P3C-MAP-peptide-stimulated peripheral blood mononuclear cells (PBMCs) from horses, chronically infected with EIAV, had memory CTL (CTLm) similar to that of PBMCs stimulated with either the minimal CTL epitopes, longer peptides containing the CTL epitopes, or EIAV. The stimulated CTL lysed EIAV-in...
Yilmaz H, Helps CR, Turan N, Uysal A, Harbour DA.The nucleoprotein of Borna disease virus (BDV-p40) was produced in a Baculovirus expression system using sf9 cells. The purity and specificity of the recombinant p40 was confirmed by SDS-PAGE and immunoblotting. The recombinant p40 was used in an ELISA to screen horse sera in Turkey. For this, 323 horses from selected cities in the Marmara region of Turkey were examined clinically and serum was collected from each. All horses were clinically healthy except for a few with wounds on the skin. Antibodies to BDV were detected in the sera of 82 (25%) of 323 horse sera. Six sera were selected that h...
van der Meulen KM, Nauwynck HJ, Pensaert MB.Equine herpesvirus-1 (EHV-1) is an important pathogen of horses, causing abortion and nervous system disorders, even in vaccinated animals. During the cell-associated viremia, EHV-1 is carried by peripheral blood mononuclear cells (PBMC), mainly lymphocytes. In vitro, monocytes are the most important fraction of PBMC in which EHV-1 replicates, however, mitogen stimulation prior to EHV-1 infection increases the percentage of infected lymphocytes. The role of the cell cycle in viral replication and the role of cluster formation in cell-to-cell transmission of the virus were examined in mitogen-s...
Macintyre AR, Dixon JB.To determine whether living hydatid tissue can, like hydatid fluid, regulate leukocyte growth, T-cell, B-cell, and macrophage lines were cocultured with protoscoleces of Echinococcus granulosus and their growth was compared with that of control cultures by thymidine uptake estimates and chemiluminescent assays of cell number. Protoscoleces supported mitosis of IL-1-deprived D10 T cells, but did not increase D10 count. The action of protoscoleces was affected by the species and organ of their origin and the length of time in culture. Unusually marked mitotic reaction, unaffected by parasite age...
Dunowska M, Meers J, Johnson RD, Wilks CR.Representational difference analysis (RDA) was used to compare gene expression in equine mononuclear cells either infected with equine herpesvirus-2 (EHV-2) or adsorbed with inactivated EHV-2. Seven clones identified in non-infected cells after three rounds of selective subtraction and enrichment for differentially expressed genes contained sequences homologous to equine monocyte chemoattractant protein 1 (MCP-1). This suggested that EHV-2 may down-regulate MCP-1 transcription in infected cells. These findings correlate well with similar findings described for human cytomegalovirus and support...
Marsh AE, Hyun C, Barr BC, Tindall R, Lakritz J.Equine protozoal myeloencephalitis (EPM), caused by a protozoal parasite infection of the central nervous system, is the most commonly diagnosed neurologic disease of horses in North America. In specific regions of the United States approximately 50% of the horse population is seropositive to Sarcocystis neurona. However, not all seropositive horses develop clinical signs. Detailed clinical examination, along with cerebrospinal fluid antibody evaluation are often used to diagnose EPM. Postmortem evaluation of the brain stem and spinal cord for histopathologic lesions compatible with nonsuppura...
Dixon S, Haswell M, Harrington D, Sutcliffe IC.We have investigated the surface localisation of the phosphotransferase system protein HPr in the equine pathogen Streptococcus equi subsp. equi using immunogold localisation and transmission electron microscopy. Like the LppC acid phosphatase lipoprotein, a reference surface antigen, the S. equi HPR could be clearly detected on the surfaces of intact cells. This study is consistent with previous reports that some streptococcal HPr is cell surface associated and suggests that the extracytoplasmic mobilisation and transfer of phosphate groups by streptococci warrant further investigation.
Bjöersdorff A, Bagert B, Massung RF, Gusa A, Eliasson I.We report the isolation and partial genetic characterization of two equine strains of granulocytic Ehrlichia of the genogroup Ehrlichia phagocytophila. Frozen whole-blood samples from two Swedish horses with laboratory-verified granulocytic ehrlichiosis were inoculated into HL-60 cell cultures. Granulocytic Ehrlichia was isolated and propagated from both horses. DNA extracts from the respective strains were amplified by PCR using primers directed towards the 16S rRNA gene, the groESL heat shock operon gene, and the ank gene. The amplified gene fragments were sequenced and compared to known seq...
Tanaka Y, Taniyama H, Tsunoda N, Shinbo H, Nagamine N, Nambo Y, Nagata S, Watanabe G, Herath CB, Groome NP, Taya K.Immunolocalization of the inhibin (a) and inhibin/activin (beta3A and betaB) subunit proteins in equine fetal testes was investigated to determine the ability of the fetal testis to produce inhibins at 120, 150, 200, and 250 days of gestation. In addition, concentrations of immunoreactive (ir)-inhibin, inhibin pro-aC, and inhibin A in both the maternal and fetal circulation were measured. It was found that plasma concentrations of ir-inhibin, inhibin pro-alphaC, and inhibin A were much higher (P < .05) in the fetal than in the maternal circulation at any stage of gestation examined. Similar...
Inagaki M, Kikuchi M, Orino K, Ohnami Y, Watanabe K.Lactoferrin with a molecular mass of 80 kDa was purified from equine seminal plasma by heparin-Agarose affinity chromatography and Sephacryl S-200 gel filtration. Purified lactoferrin was found to be highly homogeneous on the bases of its migration as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and of the monospecificity of rabbit antibodies to the purified protein in immunoblotting of seminal plasma proteins. A sandwich enzyme-linked immunosorbent assay was developed for quantifying lactoferrin in equine seminal plasma. Seminal plasma lactoferrin concentrations ...
Szeredi L, Makrai L, Dénes B.The first objective of this study was to develop an immunohistochemical procedure for rapid detection of Rhodococcus equi in impression smears from affected organs of foals on postmortem examination. The second aim was to demonstrate whether R. equi can be detected in smears of tracheal exudates collected from the same foals using an immunohistochemical method. Impression smears and cryostat and paraffin-embedded sections were made from the lungs and mediastinal lymph nodes of three foals (A, B and C) that had died of respiratory disease caused by R. equi, and also from the caudal mesenteric l...
Van Wulfen KK, Bowker RM.To localize substance P (SP) and neurokinin A (NKA) and their receptors in the insertion of the distal sesamoidean impar ligament (DSIL), deep digital flexor tendon (DDFT), and dorsal hoof wall of healthy feet of horses. Methods: 18 healthy feet from horses. Methods: Samples from the dorsal hoof wall and insertion of the DSIL and DDFT of 10 feet were processed for immunocytochemical analysis, using rabbit polyclonal antisera raised against SP and NKA. Tissue sections from 8 feet were incubated with 125-labeled SP to localize tachykinin receptors and their specificity and with control solutions...
Lucchesi PM, Parma AE, Arroyo GH.Horses infected with Leptospira present several clinical disorders, one of them being recurrent uveitis. A common endpoint of equine recurrent uveitis is blindness. Serovar pomona has often been incriminated, although others have also been reported. An antigenic relationship between this bacterium and equine cornea has been described in previous studies. A leptospiral DNA fragment that encodes cross-reacting epitopes was previously cloned and expressed in Escherichia coli. Results: A region of that DNA fragment was subcloned and sequenced. Samples of leptospiral DNA from several sources were a...
Morotomi M, Yuki N, Kado Y, Kushiro A, Shimazaki T, Watanabe K, Yuyama T.Lactobacillus equi sp. nov. is described on the basis of 18 strains isolated as one of the predominant intestinal lactobacilli from horse faecal specimens. These 18 strains were isolated from 10 horses of 6 different farms out of 20 horses of 10 farms examined. They were gram-positive, facultatively anaerobic, catalase-negative, non-spore-forming, non-motile, lactic-acid-homofermentative rods. The DNA G+C content was 38.9+/-0.8 mol %. DNA-DNA hybridization failed to associate these strains closely with any of the validly described type strains used. Analysis of the 16S rRNA gene sequence of re...
Mott J, Muramatsu Y, Seaton E, Martin C, Reed S, Rikihisa Y.Upon ingestion of adult aquatic insects, horses developed clinical signs of Potomac horse fever, and Neorickettsia risticii was isolated from the blood. 16S rRNA and 51-kDa antigen gene sequences from blood, isolates, and caddis flies fed to the horses were identical, proving oral transmission of N. risticii from caddis flies to horses.
Hamm D, Willeford KO, White G, Reed SM, Hamm J.Suppurative lower airway disease is a common debilitating disease in performance horses and, while rarely fatal, is often recalcitrant to conventional therapy. A variety of treatments have been used to combat this condition and we conducted two types of studies to determine if caprine serum fraction--immunomodulator (CSFI), a nonspecific immunomodulator, improved recovery from lower respiratory disease. Two dose response studies were performed to ascertain the efficacy of CSFI. Horses were maintained daily on conventional antibiotic therapy. Respiratory tract exudate, nasal discharge, dyspnoea...
Mungall BA, Pollitt CC.Cultured equine lamellar hoof explants secrete the pro-enzymes matrix metalloproteinase-2 (MMP-2, 72 kDa) and MMP-2 (92 kDa). Untreated explants remained intact when tested on a calibrated force transducer, but when treated with an MMP activator, developed "in-vitro laminitis", separating at the dermal-epidermal junction. Explants treated with the bacterial protease thermolysin separated dose-dependently; this was accompanied by activation of both MMP-2 and -9. Thermolysin-mediated MP activation did not occur in a cell-free system and was not inhibited by the addition of the MMP inhibitor and ...
Ellison SP, Omara-Opyene AL, Yowell CA, Marsh AE, Dame JB.A gene encoding a major 29 kDa surface antigen from Sarcocystis neurona, the primary causative agent of equine protozoal myeloencephalitis (EPM), was cloned, sequenced, and expressed as a recombinant protein. A cDNA library was prepared in the expression vector lambda ZAP from polyA+mRNA isolated from S. neurona merozoites cultivated in vitro. Random sequencing of 96 clones identified a clone of an abundant transcript having a translated amino acid sequence with 30% identity to the 31-kDa surface antigen of Sarcocystis muris cyst merozoites. Southern blot analysis indicated that the correspond...
Hannant D, O'Neill T, Ostlund EN, Kydd JH, Hopkin PJ, Mumford JA.A paresis isolate of equid herpesvirus 1 (EHV1, Ab4/8) and a plaque-purified virus derived from it (EHV1, Ab4/13), induced long-term suppression of both mitogenic and antigen-specific lymphocyte proliferations in adult outbred ponies. Peripheral blood mononuclear cells (PBMC) taken from a pony after EHV1 infection suppressed the in vitro function of normal cells but serum did not. This showed that the observed immune suppression was associated with circulating PBMC and/or their products rather than circulating soluble factors such as antigen or immune complexes. The results suggested that prod...
Boulanger P, Bannister GL, Ruckerbauer GM, Corner AH.Clinical field cases of equine infectious anemia were studied and the disease was reproduced experimentally in horses. Attempts were made to adapt the complement-fixation test to the detection of antibodies in the serum of infected animals and to the demonstration of antigens in tissue extracts.A moderate complement-fixing antibody response was demonstrated in the serum of horses shortly after primary exposure to the infectious agent. However, this reactivity was of short duration and occurred with normal as well as with infected saline tissue extracts. It was therefore concluded that this rea...
Hurcombe SD, Matthews AL, Scott VH, Williams JM, Kohn CW, Toribio RE.To determine the predictive value of serum concentrations of total protein (sTP), albumin (sAlb), and globulin (sGlob) measured by automated calorimetric assays to estimate serum immunoglobulin G (sIgG) concentrations in neonatal foals and identify failure of transfer of passive immunity when compared to turbidoimmunometric assay determinations of sIgG. Methods: Retrospective and prospective analysis of laboratory data. Methods: University tertiary care facility. Methods: Group 1 (retrospective): foals (n = 45) ≤7 days of age in which sIgG, sGlob, sAlb, and sTP concentrations were measured o...
Wobeser BK.During a widespread anthrax outbreak in Canada, miniature horses were vaccinated using a live spore anthrax vaccine. Several of these horses died from an apparent immune-mediated vasculitis temporally associated with this vaccination. During the course of the outbreak, other miniature horses from different regions with a similar vaccination history, clinical signs, and necropsy findings were found. Vaccin contre l’anthrax associé à la mort de chevaux miniatures. Durant une vaste éclosion d’anthrax au Canada, des chevaux miniatures ont été vaccinés en utilisant un vaccin à base de sp...
Bishop AE, Hodson NP, Major JH, Probert L, Yeats J, Edwards GB, Wright JA, Bloom SR, Polak JM.In recent years, distinct changes in regulatory peptides have been found in a number of gastrointestinal diseases. Grass sickness is a fatal disease of horses for which the etiology has yet to be fully ascertained. In this study, the peptide-containing nerves and ganglionic and mucosal endocrine cells of the ileum, colon and rectum were investigated in horses with sub-acute or chronic grass sickness and compared with normal controls using immunocytochemistry, at both the light and electron microscopical levels, and radioimmunoassay. A substantial loss of both peptide-containing cells and nerve...
Leegaard J, Roth A.Case history, skin tests and RAST were compared in a group (n = 40) of children with a history suggesting allergy to horses and in a group (n = 43) in whom there was no suspicion of hypersensitivity to horses. There was an agreement of 91% between case history and prick test. The same magnitude of agreement was found between case history and RAST (89%), and the agreement between RAST and prick test was 90%. The results of this investigation are clearly in contrast to earlier earlier reports, in that there was a very good correlation between prick test, RAST and case history. The results sugges...
Gilger BC.This review, which is part of the "Currents in One Health" series, describes the importance of the study of immune-mediated ocular disease in the development of innovative therapeutics, such as cell and gene therapy for the eye. Recent examples of cell and gene therapy studies from the author's laboratory are reviewed to emphasize the importance of One Health initiatives in developing innovative therapies for ocular diseases. Spontaneous immune-mediated corneal disease is common in horses, cats, dogs, and humans. Autologous bone marrow-derived mesenchymal stem cells (BM-MSCs) injected subconju...
Keller LE, Tait Wojno ED, Begum L, Fortier LA.Infiltration of cluster of differentiation (CD) 3 (CD3) T cells into the synovium and synovial fluid occurs in most patients with posttraumatic osteoarthritis. During disease progression, proinflammatory T helper 17 cells and anti-inflammatory regulatory T cells infiltrate the joint in response to inflammation. This study aimed to characterize the dynamics of regulatory T and T helper 17 cell populations in synovial fluid from equine clinical patients with posttraumatic osteoarthritis to determine whether phenotype and function are associated with potential immunotherapeutic targets. An imbala...
Tzipori S, Withers M, Hayes J, Robins-Browne R, Ward KL.Equine small intestinal brush-border membranes, from 40 adult horses were tested in vitro for the presence of receptors for the Escherichia coli adhesive antigens K88ab, K88ac and K99. Only K88-positive strains of E. coli adhered strongly to horse brush-border membranes. In contrast, a K88-negative mutant strain J2, 2 K99-positive strains and 3 E. coli strains isolated from foals failed to adhere to horse brush-border membranes. Purified K88ac pili when reacted with equine brush-border membranes inhibited to a great extent the adhesion of K88-positive E. coli. Similarly, K88-positive E. coli p...
Winder NC, von Fellenberg R.The peroxidase anti-peroxidase technique was used to demonstrate free and intracellular immunoglobulin (Ig) within the lungs of 23 horses with chronic small airway disease. Histologically, all the horses had chronic bronchiolitis; however, the lesions varied in degree from mild in eight horses, to moderate in nine horses and severe in six horses. Lungs from three horses which had no gross or histopathological lesions were used as controls. In comparison with control horses, horses with mild chronic bronchiolitis had increased numbers of Ig A-containing and non-immunoglobulin staining cells aro...
Kovacs S, Racz B, Sotonyi P, Bakos Z.Atrial fibrillation is the most common arrhythmia in horses causing poor performance. The role of pulmonary vein triggers in the pathogenesis has been identified in horses. Ablation methods have been investigated, but the available information on anatomical, histological and immunohistochemical assessment of the pulmonary vein ostia and the conduction system of the myocardial sleeve is still limited. Objective: The aim of the study was to describe the morphological properties of the myocardial sleeve in healthy horses. Methods: Cross-sectional. Methods: Eighty-three equine hearts were dissecte...
Elsheikha HM, Mansfield LS.Sarcocystis neurona has become recognized as the major causative agent of equine protozoal myeloencephalitis (EPM) in the Americas. At least 3 pathogenic species of Sarcocystis, including S. neurona, can be isolated from opossums. Methods are needed to ascertain whether these isolates are viable and capable of causing infections. In this study, the nuclear stain propidium iodide (PI) was used to differentiate between live (viable) and heat-killed (nonviable) S. neurona sporocysts. PI was excluded by live sporocysts but penetrated compromised sporocyst membrane and stained sporozoite nuclei of ...
Høie S, Fossum K.An agar diffusion method using microtiter plates was used to detect antibodies to the DNases produced by Staphylococcus aureus, S. intermedius, and S. hyicus. Antibodies to DNase from S. aureus were demonstrated in most of the sera from the species investigated, except dogs, only 11% of whose sera were positive. Positive titers to S. intermedius DNase were found in 84% of deg sera, 61% of Icelandic pony sera, 41% of pig sera, 21% of human sera, and 20% of cow sera but in only 2 and 4% of goat and sheep sera, respectively. Although antibodies to DNase from S. hyicus were not found in sera from ...
Ozaki H, Shimizu-Nei A, Sugita S, Sugiura T, Imagawa H, Kida H.To provide information on the antigenic variation of the hemagglutinins (HA) among equine H 3 influenza viruses, 26 strains isolated from horses in different areas in the world during the 1963-1996 period were analyzed using a panel of monoclonal antibodies recognizing at least 7 distinct epitopes on the H 3 HA molecule of the prototype strain A/equine/Miami/1/63 (H 3 N 8). The reactivity patterns of the virus strains with the panel indicate that antigenic drift of the HA has occurred with the year of isolation, but less extensively than that of human H 3 N 2 influenza virus isolates, and diff...
Xuan X, Igarashi I, Tanaka T, Fukumoto S, Nagasawa H, Fujisaki K, Mikami T.A latex agglutination test (LAT) using recombinant equi merozoite antigen 1 (EMA-1) for the detection of antibodies to Babesia equi was developed. The LAT was able to differentiate very clearly between sera from B. equi-infected horses and sera from Babesia caballi-infected horses or from normal horses. The LAT results were identical to those of a previously developed enzyme-linked immunosorbent assay. These results indicate that LAT using recombinant EMA-1 might be very useful as a routine screening method for the diagnosis of B. equi infection.
Magnuson NS, Perryman LE, Wyatt CR, Ishizaka T, Mason PH, Namen AE, Banks KL, Magnuson JA.Peripheral blood mononuclear cells (PBMC) from 14 foals with hereditary severe combined immunodeficiency (SCID) were studied to determine the extent of lymphocyte differentiation that occurs in this disorder. PBMC from all 14 horses had the morphologic characteristics of large granular lymphocytes (LGL). Cells from only one of 14 horses were responsive to phytolectin stimulation in a standard blastogenesis assay; however, PBMC from all 14 horses proliferated in continuous culture in the presence of partially purified interleukin 2. Furthermore, there were differences in the growth patterns of ...
Jenvey C, Caraguel C, Howarth GB, Riley CB.Prior to the start of endogenous production of immunoglobulins (Igs), absorption of maternal Igs is important to protect against pathogens in the early neonatal period. It is possible that mare- or foal-associated factors may influence neonatal IgA concentrations. Objective: The temporal relationships among serum and milk IgA concentrations in Thoroughbred mare-foal pairs were explored to determine if periparturient mare- and foal-associated factors contribute to the prediction of foal serum IgA concentrations. Methods: Blood and milk samples as well as complete veterinary records, were collec...
Liu IK, Turner JW, Van Leeuwen EM, Flanagan DR, Hedrick JL, Murata K, Lane VM, Morales-Levy MP.In this study of equids, we investigated the antibody response and the effect on the estrous cycle following a single inoculation of porcine zonae pellucidae (pZP) employing controlled-release methodology. We also investigated the use of two different water-soluble adjuvants as an alternative to oil-based adjuvants. Twenty-seven domestic mares were inoculated with various formulations of pZP and adjuvant. We showed that the anti-pZP antibodies generated as a result of the inoculations persisted for at least 43 weeks (length of the study). Of the various formulations used in the study, pZP and ...
Okumura M, Fujinaga T.To establish a sensitive and specific monoclonal antibody (MAB) against equine keratan sulfate (KS) and to develop an enzyme immunoassay for measurement of the concentration of KS in serum and synovial fluid from horses. Methods: 18 synovial fluid and 48 serum samples were obtained from clinically normal horses and horses with arthritis. Methods: BALB/c mice were immunized with chondroitinase-ABC-digested proteoglycan monomer from equine joint cartilage, and MAB were raised, using Sp2/O cells as a fusion partner. A competitive ELISA was optimized, using one of the established MAB, and KS conce...
Lancheros-Buitrago J, Rodriguez-Villamil P, Gregory J, Bastos H, Camacho CA, Caballeros JE, Cazales N, Barros E, José de Jesus Silva M, Pimentel A....Characterization of the uterine proteome before the entry of the conceptus to the uterus is essential to know the factors involved in the physiological events of gestation. The objective of the study was to compare proteomic profile of uterine fluid collected on day 5 post ovulation of cyclic and inseminated mares. Samples of endometrial secretion were recovered over 2 cycles during the fifth day post ovulation. The first cycle constituted the Cyclic group and in the following cycle, the same mares were inseminated and considered as the Inseminated group. All the samples were subjected to two-...
Barton AJ, Prow NA, Hall RA, Kidd L, Bielefeldt-Ohmann H.This report summarises the findings from a case of naturally-occurring Murray Valley encephalitis in a 2-year-old filly presenting with acute onset of depression and weakness. Serum samples tested at the onset of clinical signs were negative for Hendra and Kunjin virus antibodies, but positive for Murray Valley encephalitis virus (MVEV) using IgM-capture ELISA (1 : 300 dilution). A virus neutralisation assay performed 4 weeks later confirmed a titre of 1 : 160. Sera collected in the weeks preceding neurological signs returned a negative titre for MVEV 2 weeks prior followed by a titre ...
Elferink AJW, Entiriwaa D, Bulgarelli P, Smits NGE, Peters J.Processed milk and milk products produced from bovine milk, commonly contain β-casein A1 (βCA1) and β-casein A2 (βCA2). Since the presence of βCA1 is linked to milk intolerance and digestion problems, A2A2 milk, which only contains βCA2, is proposed as a healthier alternative. To support this health claim, the purity of A2A2-milk has to be guaranteed. In the presented study, a multiplex immunoassay, able to distinguish between βCA2 and βCA1, was developed and real-life applicability was shown on raw milk samples from genotyped A1A1, A1A2 and A2A2 cows. Because of its ability to discrim...
Barton MH, Bruce EH, Moore JN, Norton N, Anders B, Morris DD.To test efficacy of murine monoclonal, rabbit polyclonal recombinant equine or human tumor necrosis factor-alpha (rETNF or rHTNF, respectively) antibodies to inhibit native equine tumor necrosis factor (TNF) activity. Methods: 8 and 18 healthy adult horses for parts 1 and 2 of the study, respectively. Methods: In part 1, supernates from endotoxin-activated peritoneal macrophages were incubated with various dilutions of each rETNF antibody and subsequently tested for TNF activity. Serum was also obtained from a horse 1 hour after infusion with 20 ng of endotoxin/kg of body weight and was incuba...
van der Meulen KM, Nauwynck HJ, Pensaert MB.In the present study, equine herpesvirus-1 (EHV-1)-infected cells were identified in ionomycin/phorbol dibutyrate (IONO/PDB)-stimulated peripheral blood mononuclear cells (PBMC) and the mechanism by which stimulation increases the percentage of infected cells was examined. In the population of viral antigen-positive PBMC, 38.4+/-4.5% were CD5(+) T-lymphocytes (18.1+/-3.2% CD4(+) 13.6+/-1.8% CD8(+)), 18.1+/-5.4% were B-lymphocytes, 8.5+/-3.9% were monocytes and 35% remained unidentified. The role of the cell cycle in the increased susceptibility to EHV-1 upon stimulation was examined by stimula...
Hwang HJ, Han JW, Jeon H, Han JW.Lectin is an important protein in medical and pharmacological applications. Impurities in lectin derived from natural sources and the generation of inactive proteins by recombinant technology are major obstacles for the use of lectins. Expressing recombinant lectin with a tandem repeat structure can potentially overcome these problems, but few studies have systematically examined this possibility. This was investigated in the present study using three distinct forms of recombinant mannose-binding lectin from (BPL2)-i.e., the monomer (rD1BPL2), as well as the dimer (rD2BPL2), and tetramer (rD4...
Kahn SK, Cywes-Bentley C, Blodgett GP, Canaday NM, Turner-Garcia CE, Flores-Ahlschwede P, Metcalfe LL, Nevill M, Vinacur M, Sutter PJ, Meyer SC....Hyperimmune plasma raised against β-1→6-poly-N-acetyl glucosamine (PNAG HIP) mediates more opsonophagocytic killing of Rhodococcus equi (R equi) than does R equi hyperimmune plasma (RE HIP) in vitro. The relative efficacy of PNAG HIP and RE HIP to protect foals against R equi pneumonia, however, has not been evaluated. Objective: Transfusion with PNAG HIP will be superior to RE HIP in foals for protection against R equi pneumonia in a randomized, controlled, blinded clinical trial. Methods: Four hundred sixty Quarter Horse and Thoroughbred foals at 5 large breeding farms in the Un...
van Doorn DC, Kooyman FN, Eysker M, Hodgkinson JE, Wagenaar JA, Ploeger HW.Cyathostomins are considered to be the primary helminth pathogen of horses and macrocyclic lactones (ML) are the most frequently used anthelmintics. Therefore, ML resistance is a serious threat for the control of these parasites. In the present study ivermectin resistant cyathostomin L3 were in vitro selected, using a reiterative larval migration inhibition assay (rLMIA) and differentiated by reverse line blot (RLB). Larvae were obtained from two populations, one from a never treated, free-roaming horse population in the nature reserve Oostvaardersplassen (OVP) and the other from regularly ive...
du Plessis DH, van Wyngaardt W, Bremer CW.African horsesickness virus (AHSV), an important disease of equines is caused by an orbivirus. Because of the need to contain the spread of the disease, it is often essential to make a rapid diagnosis. For this purpose, an ELISA capable of detecting viral antigen in animal tissue and in cell culture fluid was developed. Immobilised F(ab')2 fragments prepared by digestion of AHSV-specific IgG with pepsin were used to trap virus from tissue homogenates or cell culture supernatant. After addition of intact IgG as detecting antibody, Staphylococcus aureus protein A labelled with horseradish peroxi...
Marr KA, Foster AP, Lees P, Cunningham FM, Page CP.The effect of antigen challenge on the state of activation of peripheral blood neutrophils from horses with chronic obstructive pulmonary disease (COPD) has been determined by measuring neutrophil superoxide anion formation. Prior to a seven-hour antigen challenge superoxide anion production by neutrophils from asymptomatic horses with COPD and normal horses in response to platelet activating factor (PAF) (with and without cytochalasin B), serum treated zymosan (STZ) and phorbol myristate acetate (PMA) was similar. Agonist-induced superoxide production by neutrophils from symptomatic COPD and ...
Sabban S, Ye H, Helm B.The interaction of IgE with its high-affinity Fc receptor (FcεRI) followed by an antigenic challenge is the principal pathway in IgE mediated allergic reactions. As a consequence of the high affinity binding between IgE and FcεRI, along with the continuous production of IgE by B cells, allergies usually persist throughout life, with currently no permanent cure available. Horses, especially race horses, which are commonly inbred, are a species of mammals that are very prone to the development of hypersensitivity responses, which can seriously affect their performance. Physiological responses ...
