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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.
Prevalence of hemagglutination-inhibition and neutralizing antibodies to arboviruses in horses of java. A study was conducted to measure the prevalence of hemagglutination-inhibition (HI) and neutralizing antibodies against two arboviruses (Chikungunya and Japanese encephalitis virus) in horses of Java, Indonesia. Blood specimens were collected from a sample of 112 horses at two stables: Pulo Mas, a racing track-horse complex, located in a residential area in North Jakarta, and Pamulang, a riding school, located in a rural environment of West Jaya. Sera were tested by the HI assay and plaque reduction neutralization test. JEV antibodies were detected by HI in 58 (52%) of the horses, while only 1...
Polyclonal antibody-based antigen-detection immunoassay for diagnosis of Trypanosoma evansi in buffaloes and horses. An enzyme-linked immunosorbent assay (ELISA) was employed for the detection of Trypanosoma evansi antigens in serum samples of field cases of buffaloes and horses in northern India. In 323 naturally infected/suspected buffaloes, circulating antigenaemia was detected in 180 (55.72%), whereas parasitaemia by wet blood smear examination was found in 62 (19.19%) only. The antigen-ELISA was positive in 47 of the 62 parasitologically proven cases and in 86 of the 116 cases with anti-trypanosome antibodies detected by ELISA. Of the 80 horses examined antigen-ELISA was positive in 45 (56.75%) sera. Th...
Cloning and expression of two genes from Babesia equi merozoites and evaluation of their diagnostic potential. High-titre equine immune sera were used to screen a lambda gt 11 expression library of Babesia equi cDNA fragments. Two cDNA clones which did not cross-hybridize to each other were studied. Both clones hybridized specifically to DNA from B. equi but not to DNA from B. caballi, B. divergens or B. ovis. Recombinant proteins were expressed as glutathione S-transferase (GST) fusion proteins with apparent molecular weights of 40 kDa and 75 kDa. Polyclonal antibodies directed against the 40 kDa and 75 kDa recombinant proteins detected native antigens of 55 kDa and 50 kDa respectively in crude lysate...
Localization of a protective epitope on a Venezuelan equine encephalomyelitis (VEE) virus peptide that protects mice from both epizootic and enzootic VEE virus challenge and is immunogenic in horses. In order to define more precisely the protective epitope encoded within the first 25 amino acids (aa) of the E2 glycoprotein of the Trinidad donkey strain of Venezuelan equine encephalomyelitis (VEE) virus, we examined the immunogenicity of smaller peptides within the first 19 aa. pep1-9 and pep3-10 elicited virus-reactive antibody, but failed to protect mice from virus challenge. Additionally, pep3-10 was identified by a competitive binding assay using overlapping peptide octamers as the putative binding site of the antipeptide monoclonal antibody (mAb) 1A2B-10. Since the E2 amino-terminal se...
Production of monoclonal antibodies in horses. Monoclonal antibodies (MAbs) have been successfully used to evaluate immune responses in horses, and to target important antigens of equine infectious agents to which protective immune responses may be directed (1–5). Most of these studies are performed with murine MAb produced by fusing spleen cells from immunized mice with an appropriate myeloma cell line, as described in Chapter 3. However, there are experimental questions for which murine MAb are not adequate. These include:
1.Definition of microbial epitopes recognized by the infected host;
2.Identification of immunodominant epito...
Immunisation of mares to control endometritis caused by Streptococcus zooepidemicus. Normal mares were immunised by the intramuscular and intrauterine administration of an antigen with adjuvant and they and unimmunised control mares were later challenged by the intrauterine instillation of pathogenic Streptococcus zooepidemicus; the response of all the mares was monitored clinically and bacteriologically for seven days. Significantly fewer S zooepidemicus were present in cervical swabs taken from the immunised mares than from the control mares (P < 0.01) and the degree of inflammation in the genital tract of the immunised mares was also significantly less (P < 0.001). Th...
Continuing prevalence of African horse sickness in Nigeria. Equine sera collected from 10 widely separated regions throughout Nigeria were tested for antibodies against African horse sickness viruses (AHSV) using a competitive enzyme-linked immunosorbent assay (ELISA). The animals sampled included imported, exotic horses, indigenous and locally cross-bred (local) horses and African donkeys. A high percentage of the sera (79.8%) were positive, confirming the continued prevalence of AHSV antibodies in Nigerian horses and donkeys.
Seroanalysis of Tyzzer’s disease in horses: implications that multiple strains can infect Equidae. A monoclonal antibody based competitive inhibition assay was used to detect antibodies in horse sera to purified flagellar antigens from distinct Clostridium piliforme isolates. Sequential absorption of hyperimmune rat serum to C. piliforme isolate E (horse-origin isolate), a positive C. piliforme-immune horse serum, and other suspected immune horse sera with unrelated bacteria or C. piliforme isolates E or isolate R1 (rat-origin isolate) alone demonstrated the specificity of this assay for C. piliforme. This specificity was associated with the inhibition of monoclonal antibody binding to C. p...
Bovine respiratory syncytial virus antibodies in non-bovine species. To study the role of non-bovine species in the epidemiology of bovine respiratory syncytial virus (RSV) infections, sera obtained from 9 non-bovine animal species and from humans were examined for bovine RSV specific antibodies. Sera were mainly from animals and humans which had been in contact with cattle. Forty sera of each species were tested in an RSV specific whole virus ELISA as well as in a peptide based ELISA, that was developed to measure antibodies specific for bovine RSV. Antibodies directed against RSV were detected in over 50% of sera obtained from sheep, goat, cattle and human be...
Enzyme-linked immunosorbent assay for myosin heavy chains in the horse. The content in slow and fast myosin heavy chains (MHC 1 and MHC 2) of 5 equine muscles was determined using an enzyme-linked immunosorbent assay. The results obtained with this immunoenzymatic method were compared with complementary techniques: electrophoresis and immunohistochemistry. Slices of masseter, diaphragm, tensor faciae latae, semitendinosus and cutaneus trunci were obtained from a 12-year-old saddle horse after slaughter. Muscular proteins were specifically extracted to be analysed by ELISA. The technique used 2 complimentary monoclonal antibodies (MAb). MAb 1 was prepared from a hu...
A type-specific serological test to distinguish antibodies to equine herpesviruses 4 and 1. We describe a type-specific ELISA, which distinguishes antibody to equine herpesvirus 4 (EHV4; equine rhinopneumonitis) and EHV1 (equine abortion virus) thereby identifying horses that have been infected with either or both of these antigenically related viruses. The antigens used are parts of the EHV4 and EHV1 glycoprotein G (gG) homologues expressed in E. coli as fusion proteins [Crabb and Studdert, 1993: J Virol 67: 6332-6338). The expressed proteins comprise corresponding regions of the gG molecules that are highly divergent and encompass strong, typespecific epitopes. Plasma samples from ...
Response of ponies to adjuvanted EHV-1 whole virus vaccine and challenge with virus of the homologous strain. Five yearling ponies were vaccinated with inactivated Equid herpesvirus type 1 (EHV-1) in Freund's complete adjuvant as a double emulsion and revaccinated 6 weeks later with EHV-1 in Freund's incomplete adjuvant. These ponies and three age-matched controls were challenged intra-nasally after a further 6 weeks with homologous live virus and monitored clinically, biologically and serologically. After challenge, clinical signs were mild in both groups. No cell-associated viraemias were detected in vaccinated ponies. Vaccination induced high levels of complement-fixing (CF) and virus-neutralizing ...
Generic immunoassay of corticosteroids with minimum pre-treatment of urine samples. A generic, rapid and sensitive enzyme linked immunosorbent assay (ELISA) test has been developed which allows large-scale simultaneous testing of synthetic corticosteroids viz., flumethasone, dexamethasone and betamethasone. This assay can be directly applied to diluted urine samples (1 + 9) without hydrolysis of glucuronide or sulfate conjugates or any other treatment of samples. The polyclonal antibody was obtained by immunizing sheep with a flumethasone derivative linked to human serum albumin. This polyclonal antibody displayed high-reactivity with several synthetic corticosteroids whilst ...
Treatment of aplastic anemia with an investigational antilymphocyte serum prepared in rabbits. The authors evaluated antilymphocyte serum prepared in rabbits (ALS-R) as an alternative to antilymphocyte serum prepared in horses (ALG-H) in the therapy of aplastic anemia. Between 1980 and 1993, 57 evaluable patients received ALS-R and prednisone +/- cyclosporine +/- androgens. Standard response criteria were used and patients were evaluated at 3 months from the start of therapy. Median age was 43 years. Disease was present for up to 2 months in 24 patients, 2-5 months in 14 patients, and 6 months or more in 19 patients. Disease was severe in 30 patients and moderate in 27. Responses occurr...
Experimental immunization of ponies with Strongylus vulgaris radiation-attenuated larvae or crude soluble somatic extracts from larval or adult stages. Protection from Strongylus vulgaris infection through immunization with radiation-attenuated third-stage larvae (L3) or crude soluble homogenates from larval or adult stages was examined. Yearling ponies raised parasite-free were divided into 3 immunization groups: radiation-attenuated L3; soluble adult somatic extracts; larval somatic extracts with excretory/secretory products (E/S) from in vitro culture; and 1 medium control group. Ponies were immunized twice; attenuated larvae were administered orally and somatic extracts or controls injected intramuscularly with adjuvant. Approximately 6 w...
Genetic and antigenic analysis of the influenza virus responsible for the 1992 Hong Kong equine influenza epizootic. An outbreak of influenza occurred among thoroughbred racehorses in Hong Kong in November-December 1992, with morbidity of 37%. All horses involved had been vaccinated against equine-1 and equine-2 influenza viruses but not against the virus responsible for the 1989 equine influenza outbreak in northern China (influenza A/equine/Jilin/89, subtype H3N8). Therefore the source and nature of the virus causing the Hong Kong outbreak was investigated. Virus isolated from a horse infected during the outbreak was used for genetic analysis. All the viral gene segments were similar to those of equine-2 (...
Structural protein relationships among eastern equine encephalitis viruses. We have re-evaluated the relationships among the polypeptides of eastern equine encephalitis (EEE) viruses using SDS-PAGE and peptide mapping of individual virion proteins. Four to five distinct polypeptide bands were detected upon SDS-PAGE analysis of viruses: the E1, E2 and C proteins normally associated with alphavirus virions, as well as an additional more rapidly-migrating E2-associated protein and a high M(r) (HMW) protein. In contrast with previous findings by others, the electrophoretic profiles of the virion proteins of EEE viruses displayed a marked correlation with serotype. The pro...
Genetic Bit Analysis: a solid phase method for typing single nucleotide polymorphisms. A new method for typing single nucleotide polymorphisms in DNA is described. In this method, specific fragments of genomic DNA containing the polymorphic site(s) are first amplified by the polymerase chain reaction (PCR) using one regular and one phosphorothioate-modified primer. The double-stranded PCR product is rendered single-stranded by treatment with the enzyme T7 gene 6 exonuclease, and captured onto individual wells of a 96 well polystyrene plate by hybridization to an immobilized oligonucleotide primer. This primer is designed to hybridize to the single-stranded target DNA immediately...
[Epidemiology of melioidosis in China]. From 1975 to 1989, a total of 73 strains of P. pseudomallei was isolated from the water samples and the pathological samples of human and domestic animals in 13 counties and cities located different latitude from four provinces Qiong, Yue, Gui and Xiang in China. Serological investigation demonstrated that the geographical distribution of the organism had a significant correlation with the positive rate of antibodies against P. pseudomallei and the native foci of the organisms distributed over the southern subtropical zone and the edge of tropical zone in Qiong, Yue and Gui. In endemic areas, ...
Titers to Leptospira species in horses in Alberta. Sera from horses in Alberta, submitted to Agriculture and Agri-Food Canada for routine testing for equine infectious anemia from January 1987 to June 1989, were tested for antibody against 13 serovars of pathogenic Leptospira spp., using the microscopic agglutination test. The purpose of the study was to investigate the prevalence of serum titers to those serovars in horses in Alberta, and to analyze the associated risk factors. Descriptive statistics were compiled and logistic regressions were computed. Titers to L. interrogans serovars icterohaemorrhagiae, bratislava, copenhageni, and autumn...
Eastern equine encephalomyelitis virus in relation to the avian community of a coastal cedar swamp. Eastern equine encephalomyelitis virus (EEEV) is perpetuated in eastern North America in a mosquito-wild bird maintenance cycle that involves Culiseta melanura (Coquillett) as the principal enzootic vector and passerine birds as the primary amplifying hosts. We examined the role of birds in the EEEV cycle at a site in southern New Jersey where EEEV cycles annually at high levels. Birds and mosquitoes were sampled during three epiornitics and one season of limited virus activity. We examined antibody prevalence in birds in relation to eight physical and natural history characteristics. Our goal...
Monoclonal antibodies to equine arteritis virus proteins identify the GL protein as a target for virus neutralization. Monoclonal antibodies (MAbs) to equine arteritis virus (EAV) proteins were produced and characterized. The protein specificities of eight MAbs were determined definitively by immunoprecipitation of EAV proteins expressed from vaccinia virus recombinants (VVRs). Included were two new VVRs produced for this study, expressing the M and the GL proteins, respectively. Three MAbs were determined to be N-specific and five MAbs recognized the GL protein. One GL-specific MAb, 17F5, of the IgA class, efficiently neutralized EAV infectivity. In competitive binding assays (CBAs), the N-specific MAbs defin...
Proteolytic processing of the replicase ORF1a protein of equine arteritis virus. To study the proteolytic processing of the equine arteritis virus (EAV) replicase open reading frame 1a (ORF1a) protein, specific antisera were raised in rabbits, with six synthetic peptides and a bacterial fusion protein as antigens. The processing of the EAV ORF1a product in infected cells was analyzed with Western blot (immunoblot) and immunoprecipitation techniques. Additional information was obtained from transient expression of ORF1a cDNA constructs. The 187-kDa ORF1a protein was found to be subject to at least five proteolytic cleavages. The processing scheme, which covers the entire OR...
Diagnosis of African horsesickness. African horsesickness (AHS) is a very serious, non-contagious disease of horses and other solipeds caused by an arthropod-borne orbivirus of the family Reoviridae. The epizootic nature of the disease makes rapid, accurate diagnosis of AHS absolutely essential. Currently, diagnosis of AHS is based on typical clinical signs and lesions, a history consistent with vector transmission and confirmation by laboratory detection of virus and/or anti-AHS virus antibodies. The clinicopathologic presentation of AHS, current and next generation laboratory diagnostic methods are discussed.
Evaluation of a monoclonal antibody-based colony blot test for rapid identification of virulent Rhodococcus equi. We recently generated a monoclonal antibody immunoglobulin G1 (MAb 10G5), which can recognize 15- to 17-kDa antigens, virulence-associated antigens of Rhodococcus equi, and developed a colony blot enzyme-linked immunosorbent assay with MAb 10G5 for the rapid identification of virulent R. equi. In this epidemiologic study, we evaluated the results of the colony blot test in the identification of virulent isolates of R. equi from feces of horses and soil and compared them with those from a conventional procedure (plasmid profiles of isolates by agarose gel electrophoresis). Environmental isolate...
The immunology of companion animals: reagents and therapeutic strategies with potential veterinary and human clinical applications. There is now a wide range of immunological reagents that can be used in the diagnosis and treatment of diseases in the companion animals (dogs, cats and horses). Many of these diseases are the veterinary equivalents of human conditions, and may therefore provide good models to study basic pathogenic mechanisms.
African horsesickness: pathogenesis and immunity. African horsesickness (AHS) is a serious, non-contagious disease of horses and other solipeds caused by an arthropod-borne orbivirus of the family Reoviridae. In horses, AHS causes three distinct clinicopathologic syndromes, the pulmonary, cardiac and fever forms of the disease. Recent work has shown that the primary determinant of the form of disease expressed by naive horses is the virulence of the virus inoculum. Horses which recover from AHS exhibit solid humoral immunity against homologous challenge. Protective antibodies appear to be directed towards neutralizing epitopes on AHS virus VP...
Characterization of the antiphagocytic activity of equine fibrinogen for Streptococcus equi subsp. equi. The antiphagocytic property of equine fibrinogen for Streptococcus equi subsp. equi strain CF32 was examined in vitro. The results of bactericidal assays demonstrated that the presence of fibrinogen enhanced the ability of overnight and early log-phase cultures of strain CF32 to resist killing by equine neutrophils by 12-fold and seven-fold, respectively (p > 0.01). In addition, fibrinogen-coated bacteria treated with fibrinogen specific F(ab')2 fragments were 32% more susceptible to killing by equine neutrophils after opsonization in serum (p > 0.05), indicating that specific epitopes o...
Characterization of monoclonal antibodies specific for equine homologues of CD3 and CD5. Two monoclonal antibodies (mAb), UC F6G-3 and UC F13C-5, were characterized as being specific for the apparent equine homologues of CD3 and CD5, respectively. Both antibodies exhibited characteristics of pan-T-lymphocyte markers based upon immunohistology and two-colour flow cytometry. UC F6G-3 precipitated a complex of proteins (up to seven) with molecular weights ranging from 18,000 to 42,000, similar to the human and murine CD3 complex. Upon further dissociation of the precipitated complex, two proteins were identified with molecular weights of 22,000 and 27,000. Immobilized UC F6G-3 was ef...
