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Topic:Antibodies
Antibodies in horses are specialized proteins produced by the immune system in response to foreign substances, known as antigens. These substances can include pathogens such as bacteria, viruses, and parasites. Antibodies function by recognizing and binding to specific antigens, thereby neutralizing them or marking them for destruction by other immune cells. In equine health, antibodies are integral to both natural immune responses and those induced by vaccinations. The study of antibodies in horses encompasses their production, diversity, and role in disease resistance and management. This page gathers peer-reviewed research studies and scholarly articles that explore the generation, function, and implications of antibodies in equine immunology and disease control.
Distribution of fast myosin heavy chain-based muscle fibres in the gluteus medius of untrained horses: mismatch between antigenic and ATPase determinants. The distribution of muscle fibres classified on the basis of their content of different myosin heavy chain (MHC) isoforms was analysed in muscle biopsies from the gluteus medius of adult untrained horses by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies and standard myofibrillar ATPase (mATPase) histochemistry. Percutaneous needle biopsies were taken at 3 depths (20, 40 and 60 mm) from 4 4-y-old Andalusian stallions. The percentage of 'pure' I MHC fibres increased whereas that for pure IIX MHC fibres decreased from the most superficial to the deepest sampling sit...
Detection of equine antibodies to babesia caballi by recombinant B. caballi rhoptry-associated protein 1 in a competitive-inhibition enzyme-linked immunosorbent assay. A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was developed for detection of equine antibodies specific for Babesia caballi. The assay used recombinant B. caballi rhoptry-associated protein 1 (RAP-1) and monoclonal antibody (MAb) 79/17.18.5, which is reactive with a peptide epitope of a native 60-kDa B. caballi antigen. The gene encoding the recombinant antigen was sequenced, and database analysis revealed that the gene product is a rhoptry-associated protein. Cloning and expression of a truncated copy of the gene demonstrated that MAb 79/17.18.5 reacts with the C-termina...
Detection of equine herpesvirus types 2 and 5 (EHV-2 and EHV-5) in Przewalski’s wild horses. In blood samples of seven captive equid species from four German zoos EHV-1 specific antibodies were detected in 76% and EHV-4 specific antibodies in 73% of the 55 animals, whereas 93% were tested positive for EHV-2 and EHV-5, respectively. In only one blood sample from a Przewalski's wild horse EHV-4 DNA was amplified by PCR. From seven Przewalski's wild horses EHV-2, and from another one EHV-5 was isolated by cocultivation. The identity of the virus isolates was verified by PCR and restriction enzyme digestion.
Antigenic profile of African horse sickness virus serotype 4 VP5 and identification of a neutralizing epitope shared with bluetongue virus and epizootic hemorrhagic disease virus. African horse sickness virus (AHSV) causes a fatal disease in horses. The virus capsid is composed of a double protein layer, the outermost of which is formed by two proteins: VP2 and VP5. VP2 is known to determine the serotype of the virus and to contain the neutralizing epitopes. The biological function of VP5, the other component of the capsid, is unknown. In this report, AHSV VP5, expressed in insect cells alone or together with VP2, was able to induce AHSV-specific neutralizing antibodies. Moreover, two VP5-specific monoclonal antibodies (MAbs) that were able to neutralize the virus in a ...
Effect of exercise on the immune response of young and old horses. To compare exercise-induced immune modulation in young and older horses. Methods: 6 young and 6 aged horses that were vaccinated against equine influenza virus. Methods: Venous blood samples were collected for immunologic assessment before and immediately after exercise at targeted heart rates and after exercise for determination of plasma lactate and cortisol concentrations. Mononuclear cells were assayed for lymphoproliferative responses and incubated with interleukin-2 (IL-2) to induce lymphokine-activated killer (LAK) cells. Antibodies to equine influenza virus were measured. Results: Olde...
Serologic testing of horses for granulocytic ehrlichiosis, using indirect fluorescent antibody staining and immunoblot analysis. To diagnose granulocytic ehrlichiosis in horses, compare results of indirect fluorescent antibody (IFA) staining procedures with those of immunoblot analysis, and compare serologic test findings with polymerase chain reaction (PCR) results. Methods: 69 horses with high rectal temperatures (> or = 39 C) and lethargy, anorexia, or limb edema. Methods: 43 convalescent serum samples obtained from 38 horses 2 to 18 weeks after onset of illness were analyzed by use of immunoblot procedures and IFA staining methods, using the NCH-1 or BDS ehrlichial strains. Blood samples from 69 acutely ill horse...
The seroprevalence of equine trypanosomosis in the pantanal. Since little information is available on the epizootiological status of Trypanosoma evansi in South America and particularly Brazil, we evaluated equine serum samples collected in 1993, 1994, 1995 and 1997 for the presence of antibodies against this trypanosome species. Our study shows corroborative evidence about the correlation among high T. evansi seroprevalence and the rainy season in the Pantanal, Brazil. The higher seroprevalence was 79.2% in horses from a ranch located in the Nhecolândia sub-region in 1994 and the lower 5.8% in animals from the same ranch in 1997. No seroprevalence was...
Isolation and characterization of a novel Forssman active acidic glycosphingolipid with branched isoglobo-, ganglio-, and neolacto-series hybrid sugar chains. Equine kidney and spleen contain a Forssman active glycosphingolipid, and the structure of this glycolipid has been reported to be that of a globopentaosylceramide (GalNAcalpha-1,3GalNAcbeta-1,3Galalpha-1, 4Galbeta-1,4Glcbeta-1,1'Ceramide). We found that equine kidney contains several other anti-Forssman antibody-reactive glycosphingolipids. One of these acidic Forssman active glycosphingolipids was isolated and characterized by means of NMR, mass spectrometry, permethylation studies, and TLC-immunostaining. This glycolipid contains three moles of galactose, one mole of glucose, three moles of...
Purification and biochemical characterization of equine pulmonary surfactant protein D. To characterize surfactant protein D (SP-D) isolated from bronchoalveolar lavage fluid (BALF) of healthy horses. Methods: BALF from 10 Thoroughbreds (5 males, 5 females; 26 to 40 months old) without history or clinical signs of respiratory tract disease. Methods: BALF was obtained and centrifuged at 33,000 X g. The supernatant was applied to a mannose-Sepharose 6B affinity column in the presence of calcium, and the bound protein fraction was analyzed by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis; amino acid composition was determined and partial seque...
A 105- to 94-kilodalton protein in the epididymal fluids of domestic mammals is angiotensin I-converting enzyme (ACE); evidence that sperm are the source of this ACE. SDS-PAGE analysis of luminal fluid from the ram testis and epididymis revealed a protein of about 105 kDa in the fluid in the caput epididymal region. The molecular mass of this fluid protein shifted from 105 kDa to 94 kDa in the distal caput epididymidis and remained at 94 kDa in the lower regions of the epididymis. The possible sperm origin of this protein was suggested by the decrease in intensity of a 105-kDa compound on the sperm plasma membrane extract and by its total disappearance from the fluid of animals with impaired sperm production caused by scrotal heating. The 94-kDa protein was...
[Detection of equine arteritis virus (EAV) in stallions–a contribution to the improvement of EAV diagnosis]. Serum samples from 72 stallions were examined for the occurrence of antibodies against equine arteritis virus, of which 41 animals (57%) were found to be positive. 32 of the seropositive stallions were then screened for persistent EAV infection, before and after the breeding season. Semen samples were investigated by RT-PCR followed by dot blot hybridization and nested PCR, and by virus isolation on cell cultures as well. The carrier state was virologically confirmed in 11 of 32 stallions (34%) during the first and in 9 of 20 (45%) during the second investigation. RT-PCR followed by confirmato...
Detection of Trichinella infection in slaughter horses by ELISA and western blot analysis. In order to determine the presence of Trichinella infections in horses slaughtered at an abattoir in Mexico, 147 serum samples were examined by two immunoenzymatic methods. Specific antibodies were detected by ELISA in 7% of the serum samples at a dilution 1:400 and in 10% at lower dilutions (1:20, 1:40) using Trichinella spiralis muscle larvae (ML) excretory/secretory (E/S) products. Serum samples from four naturally infected horses (confirmed by direct methods) gave negative O.D. values in an ELISA at a 1:400 dilution and only two of them were positive at a 1:20 and 1:40 dilutions. Serum sam...
A particulate viral protein vaccine reduces viral load and delays progression to disease in immunized ponies challenged with equine infectious anemia virus. 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. ...
Detection of antibodies to equine arteritis virus by enzyme linked immunosorbant assays utilizing G(L), M and N proteins expressed from recombinant baculoviruses. Indirect enzyme linked immunosorbant assays (ELISAs) utilizing the three major structural proteins (M, N, and G(L)) of equine arteritis virus (EAV) expressed from recombinant baculoviruses were developed. A large panel of sera collected from uninfected horses, and from animals experimentally and naturally infected with EAV or vaccinated with the modified live virus vaccine against equine viral arteritis, were used to characterize the humoral immune response of horses to the three major EAV structural proteins. The data suggest that the M protein was the major target of the equine antibody resp...
Improved hepatic and pancreatic localisation of the equine alpha-1-proteinase inhibitor family of serpins using an antigen enhancement technique and a monoclonal antibody. Equine alpha-1-proteinase inhibitor (API) consists of three, occasionally four, serum glycoproteins. This study investigated the immunohistochemical localisation of equine API in paraformaldehyde fixed, paraffin embedded equine tissue samples of liver, lung, stomach, pancreas, jejunum and colon in five horses using affinity purified sheep polyclonal and protein A purified mouse monoclonal antibodies, whose specificities were verified by Western blotting. Exposing tissue sections to boiling citrate buffer greatly enhanced antigen recovery and improved immunostaining with both antibodies, result...
Diagnosis and prevention of equine infectious diseases: present status, potential, and challenges for the future. The frequent transfers of horses, whether on a permanent or temporary basis, make strict control of infectious diseases essential. Such control needs a reliable and rapid means to accurately diagnose the relevant diseases. Indirect diagnosis based on antibody detection remains certainly the best method to secure the epidemiologic surveillance of the diseases at regional, national, or even world level, while direct diagnosis is the only way to diagnose a new outbreak. New diagnostic methods resulting from advances in biochemistry, molecular biology, and immunology are now available. As far as a...
Immunolocalization of cathepsin B in equine dyschondroplastic articular cartilage. A polyclonal antiserum raised in sheep against human cathepsin B was tested for specificity and cross-reactivity with the horse homologue by SDS-PAGE and Western blotting, prior to being used for immunolocalization of the enzyme in equine articular cartilage. In Western blots, the antiserum recognized the 30 kDa single chain and 25 kDa heavy chain of the mature enzyme in purified bovine cathepsin B, and corresponding bands at 32 and 27 kDa in equine chondrocyte and fibroblast lysates. This antiserum was then used to compare the expression and distribution of cathepsin B in normal and dyschondr...
[Demonstration of intraocular leptospira in 4 horses suffering from equine recurrent uveitis (ERU)]. Vitreous samples from 43 horses which underwent vitrectomy because of equine recurrent uveitis (ERU) were cultured for leptospires. Out of 4 vitreous samples (4/43 = 9%), leptospires could be isolated. In 3 cases, serovar grippotyphosa, and in one case, a serovar out of the serogroup Australis were identified. So for the first time, in several horses with ERU in vivo cultures of vitreous material were positive for leptospires. A strong evidence of association between leptospiral infection and uveitis is discussed for many years. In this investigation the leptospiral etiology is confirmed. Vitr...
Evaluation of equine infectious anemia virus core proteins produced in a baculovirus expression system in agar gel immunodiffusion test and enzyme-linked immunosorbent assay. Equine infectious anemia virus (EIAV) core proteins (Gag and p26) obtained from a baculovirus expression system were used in agar gel immunodiffusion (AGID) and enzyme-linked immunosorbent assay (ELISA) antigens to test seventy-six horse sera. Those sera showed false-positive reaction in AGID test using Nisseiken antigen. However, none of them showed false-positive reaction with both of the expressed antigens. The 76 horse sera were also tested by ELISA. The sera gave a high background in ELISA using Nisseiken antigen. Gag and p26 reacted strongly against positive sera from horses immunized wi...
Enzyme-linked immunosorbent assay (ELISA) for detection of anti-Trypanosoma evansi equine antibodies. The standardization of ELISA for the detection of anti-Trypanosoma evansi antibodies in naturally and experimentally infected horses is described. Bayesian analysis was used to establish the cutoff between positive and negative sera. In order to determine the assessment of the ELISA test, the results obtained were compared with those from an IFA. A relative sensibility of 98.39%, a specificity of 95.12% and a predictive value of 96.83% were determined. The standardized technique was used to evaluate the antibody production against trypanosome in an experimentally infected equine, in which the ...
Suppressant effect of human or equine rabies immunoglobulins on the immunogenicity of post-exposure rabies vaccination under the 2-1-1 regimen: a field trial in Indonesia. MAS054 Clinical Investigator Group. WHO's reference protocol for post-exposure rabies vaccination advises five intramuscular injections on days 0, 3, 7, 14, and 30; in addition, rabies immunoglobulins (RIG) must be given to serious cases of exposure (grade III severity). Some studies indicate that these immunoglobulins suppress the immunogenicity of rabies vaccine when administered according to an alternative protocol of four injections (2-1-1) on days 0, 7, and 21, which was therefore not recommended for grade III exposures. To test this effect, we conducted a multicentre study in Indonesia using three groups of subjects. One g...
Antibody selection for immunohistochemical survey of equine tissue. Immunohistochemical evaluation of equine tissue necessitates the use of antibodies reactive with cells from a heterogeneous population. Lymphoid tissues from 12 horses were fixed in Bouin's fluid, ethanol or formalin and examined for immunohistochemical reactivity with anti-equine and anti-human monoclonal antibodies (MAbs) specific for MHC Class II antigens, T and B lymphocytes, and macrophages. Only a few of the anti-equine MAbs tested were reactive with fixed, paraffin wax-embedded tissue. Anti-human MAbs expanded the desired range of reactivity and increased the consistency in different an...
Measurement of serum and synovial fluid keratan sulphate and antibody to collagen type II in equine osteoarthritis. Keratan sulphate (KS) concentration and anticollagen type II antibody levels were measured in the serum and synovial fluid (SF) of clinically normal horses and horses with osteoarthritis (OA). Serum KS in OA was significantly higher than that in normal horses, while no significant difference was found in KS levels of SF between normal and OA. Assays of antibody to collagen type II showed no significant increase in sera and SF of OA. It was suggested that levels of serum KS would be of value in the pathological detection of OA in the joint, although there was no evidence that the measurable aut...
Seroprevalence of equine herpesvirus 1 in thoroughbred foals before and after weaning. To investigate the seroprevalence of equine herpesvirus 1 in foals around weaning and after weaning on two large Thoroughbred farms using a type-specific enzyme-linked immunosorbent assay to determine exposure to infection. Methods: A longitudinal population study in groups of Thoroughbred weanling foals. Methods: Two hundred weanling Thoroughbred foals from a population of about 380 foals were enrolled on two adjacent stud farms in the Hunter Valley of New South Wales. Foals on both farms were weaned from February to May 1995 into randomly selected groups of 10 to 15 foals. Farms were selecte...
Immunohistochemical studies in equine recurrent uveitis (ERU). Despite extensive clinical research, the etiology of equine recurrent uveitis (ERU) is still unknown. After an immunologic pathogenesis was established in recurrent uveitis in humans, a similar pathogenic mechanism was assumed to exist in ERU. To investigate whether immunopathologic mechanisms are involved in ERU, 20 eyes of 15 horses with ERU were examined immunohistochemically with a T cell marker, B cell marker, and anti-major histocompatibility complex (MHC) class II antibodies. Twenty-six eyes of 20 horses were used for investigation of MHC class II antigen expression in normal equine eye...
Diagnosis and sero-epizootiology of equine herpesvirus type 1 and type 4 infections in Japan using a type-specific ELISA. Recently, a type-specific ELISA using equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4) glycoprotein Gs (gGs) was developed by Crabb and Studdert [1993]. To investigate the dissemination of EHV-1 and -4 among horses in Japan, we applied their ELISA as suitable for discriminating between EHV-1 and -4 infections serologically. Type-specificity of the ELISA was confirmed by using paired sera of infected horses with either EHV-1 or -4. Application of the ELISA to sera collected before and after the winter season of 1995-1996 from 80 racehorses revealed that 30 horses showed significant antibody...
Iron loading into ferritin by an intracellular ferroxidase. An intracellular, membrane-bound enzyme exhibiting both p-phenylenediamine oxidase activity and ferrous iron oxidase activity was isolated with the plasma membrane fraction of horse heart and studied for its ability to load iron into ferritin. The ferroxidase activity of the tissue oxidase was stimulated approximately twofold by horse spleen apoferritin, and the iron was loaded into ferritin. The loading of iron into ferritin by the tissue oxidase was inhibited by anti-horse serum ceruloplasmin antibody. The stoichiometry of iron oxidation and oxygen consumption during iron loading into ferrit...
Enzyme-linked immunosorbent assay for serological survey of equine arteritis virus in racehorses. To examine antibodies against equine arteritis virus (EAV), an enzyme-linked immunosorbent assay (ELISA) using purified virus antigen was developed. The results of ELISA were compared with those of serum neutralization (SN) tests. The ELISA absorbance values and the SN titers in sera collected weekly from EAV-infected horses showed a similar pattern. The ELISA could detect antibody to EAV in horses experimentally infected with not only a homologous virus strain, which was used as the ELISA antigen, but also a heterologous strain. Using the ELISA, serum samples collected in 1996 from racehorses...
Experimental infection of four horses with Ehrlichia phagocytophila. Four clinically healthy horses which were negative for antibodies to Ehrlichia phagocytophila, the agent of bovine ehrlichiosis, were infected experimentally with E phagocytophila-containing bovine leucocytes, administered intravenously. The horses were examined daily for four weeks, and blood samples were collected daily for cytological, haematological and biochemical examination and for a nested polymerase chain reaction (PCR). An indirect immunofluorescence test was used to determine when the horses seroconverted and the duration of positive titres. There were no abnormal clinical, haematol...
