Topic:Antisera
Antisera refer to blood serum containing antibodies against specific antigens, produced by the immune system in response to exposure to these antigens. In horses, antisera are commonly used for therapeutic and diagnostic purposes, particularly in the treatment of venomous bites or stings, and in combating infectious diseases. The production of equine antisera involves immunizing horses with a particular antigen and subsequently collecting and processing their blood to extract the serum rich in antibodies. This page compiles peer-reviewed research studies and scholarly articles that explore the production, application, and efficacy of antisera in equine medicine, as well as advancements in safety and regulatory considerations in their use.
Rat muscle acylphosphatase: purification, amino sequence, and immunological characterization. Acylphosphatase was purified from rat skeletal muscle essentially by gel filtration and high-performance ion-exchange chromatography. The complete amino acid sequence was reconstructed by using the sequence data obtained from tryptic, peptic, and S. aureus V8 protease peptides. The protein consists of 96 amino acid residues and is acetylated at the NH2-terminus. The immunological cross-reactivity of acylphosphatase from rat and horse skeletal muscle was examined by ELISA. The reaction with rabbit antiserum revealed the presence of at least five antigenic sites on rat enzyme, two of which are c...
Immunization of horses with Crotalus durissus terrificus (South American rattlesnake) venom. A comparison of four different procedures. 1. A comparative study was carried out on horses immunized with Crotalus durissus terrificus venom using four different inoculation procedures, which included the use of Freund's adjuvant, A1(OH)3 and liposomes as adjuvants. The antibody titer was assessed by enzyme linked immunosorbent assay (ELISA) and the neutralizing potency by the neutralizing median effective dose (ED50). 2. The inoculation schedule used in horses to obtain antivenom serum consisted of sc injections of a 7.5 mg venom starting dose in 5.0 ml sterile saline emulsified with an equal volume of Freund's complete adjuvant. One...
Antigenic relationships among the 47 human adenoviruses determined in reference horse antisera. Reference equine antisera to all 47 serotypes of human adenoviruses presently described have been prepared and evaluated by reciprocal neutralization and hemagglutination-inhibition tests. All tests were carried to endpoint dilutions a minimum of five times in each direction to give accurate values for homologous and heterologous antibody titers. Significant cross-reactions in the horse antisera were compared to similar data obtained from rabbit antisera. Using this analysis, major antigenic relationships exist among types 12-18-31 of subgenus A, types 7-11-14 and 34-35 of subgenus B, types 8-...
Purification of equine neutrophil lysozyme and its antibacterial activity against gram-positive and gram-negative bacteria. Lysozyme from equine neutrophil granulocytes was isolated in a pure form by fast performance liquid chromatography, i.e. ion-exchange chromatography and reversed-phase chromatography. The lysozyme lysed Micrococcus luteus, Bacillus subtilis and Staphylococcus lentus and was also bactericidal against the Gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, Bordetella bronchiseptica, and Serratia marcescens. Staphylococcus aureus and Staphylococcus epidermidis were not lysed. The lysozyme was only very slightly bactericidal for S. epidermidis and S. aureus. Equine neutrophil lysozyme ...
Isolation and partial structural characterization of an equine fibrinogen CNBr fragment that exhibits immunologic cross-reactivity with an A alpha-chain cross-linking region of human fibrinogen. Immunochemical studies of equine fibrinogen were conducted to characterize the structural basis for the immunologic cross-reactivity observed between human and equine A alpha chains when employing an antiserum to the 26K, human cyanogen bromide (CNBr) fragment, A alpha 241-476 (CNBr VIII). A 38K, equine CNBr fragment that reacts with this antiserum was isolated from CNBr-digested equine fibrinogen by Sephadex G-100 gel filtration. It was further purified by sequential hydrophobic chromatography on phenyl-Sepharose CL-4B, followed by reversed-phased (C-8) high-performance liquid chromatography ...
Immunodiffusion test for serodiagnosing subcutaneous zygomycosis. Culture filtrate antigens of Basidiobolus ranarum and Conidiobolus coronatus were analyzed by immunodiffusion (ID) with homologous rabbit antisera. B. ranarum and C. coronatus were each found to have five specific antigens. Results of tests with heterologous antisera indicated that all of the species shared at least one antigen. ID tests incorporating the specific precipitin bands as references were developed for detection of basidiobolomycosis and conidiobolomycosis. These tests were performed with sera from humans and horses with proven basidiobolomycosis and conidiobolomycosis as well as wi...
An indirect sandwich ELISA utilising F(ab’)2 fragments for the detection of African horsesickness virus. 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...
Invasive equine trophoblast expresses conventional class I major histocompatibility complex antigens. Monoclonal antibodies and alloantisera were used in an indirect immunohistochemical assay to determine the expression of class I and class II Major Histocompatibility Complex (MHC) antigens by equine placental cells and the endometrial tissues at the fetal-maternal interface. MHC class I antigens were expressed at high density on the surface of the trophoblast cells of the chorionic girdle at days 32-36, just prior to their invasion of the endometrium. The mature gonadotrophin-secreting cells of the endometrial cups, which are derived from the chorionic girdle cells, had greatly reduced levels...
Cloning and characterization of cDNAs encoding equine infectious anemia virus tat and putative Rev proteins. We isolated and characterized six cDNA clones from an equine infectious anemia virus-infected cell line that displays a Rev-defective phenotype. With the exception of one splice site in one of the clones, all six cDNAs exhibited the same splicing pattern and consisted of four exons. Exon 1 contained the 5' end of the genome; exon 2 contained the tat gene from mid-genome; exon 3 consisted of a small section of env, near the 5' end of the env gene; and exon 4 contained the putative rev open reading frame from the 3' end of the genome. The structures of the cDNAs predict a bicistronic message in ...
Methods for detection of immune-mediated neutropenia in horses, using antineutrophil serum of rabbit origin. Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils....
Characterisation of Chlamydia psittaci isolated from a horse. This paper describes the isolation and characterisation of a strain of Chlamydia psittaci obtained from a nasal swab taken from a horse with serous nasal discharge. Initial isolation was achieved in cycloheximide-treated McCoy cell monolayers. Chlamydial inclusions stained by immunofluorescence either with a rabbit antiserum raised against C. psittaci or with a monoclonal antibody directed against the genus-specific lipopolysaccharide antigen were single and compact. They did not stain with iodine or with a monoclonal antibody reactive against Chlamydia trachomatis. The agent was re-isolated i...
Rapid diagnosis of equine influenza. During an epizootic of equine influenza in Norway caused by influenza A/equine (H3N8) virus the efficacy of rapid virus diagnosis by the indirect immunofluorescence technique was evaluated. The antiserum used in the test was a polyclonal influenza A virus antiserum with reactivity directed mainly against the common nucleoprotein and matrix protein. This antiserum possessed sufficient reactivity for the detection of virus-infected exfoliated nasopharyngeal cells. Nasopharyngeal smear samples from 92 horses were examined and a positive diagnosis was obtained for 57 (62 per cent). Paired serum sa...
Cryopreservation of equine mononuclear cells for immunological studies. A rapid and simple technique for the cryopreservation and recovery of equine mononuclear cells was developed. Buffy-coat leukocytes were frozen in autologous plasma containing 10% DMSO and mononuclear cells were recovered by gradient sedimentation using a standard Ficoll-Hypaque purification procedure. The total numbers of mononuclear cells recovered from cryopreserved samples were 94%-82% of those recovered from fresh blood samples. The functional capabilities of the mononuclear cells from cryopreserved buffy coat preparations were compared with those of mononuclear cells from fresh samples b...
An immunohistochemical study of various peptide-containing endocrine cells and neurones at the equine ileocaecal junction. The ileocaecal junctions of 5 horses and 2 donkeys were examined by using antisera to the following peptides: somatostatin, glucagon, gastrin, neurotensin, vasoactive intestinal peptide (VIP), peptide histidine isoleucine (PHI), calcitonin gene-related peptide (CGRP), substance P (SP) and neuropeptide Y (NPY). Antisera to somatostatin, neurotensin and NPY demonstrated endocrine cells in the ileal- and caecal parts of the ileocaecal junction, while immunoreactivity for glucagon was demonstrated in endocrine cells of the ileal part only. Nerve cell bodies showing immunoreactivity to SP, VIP, CGR...
Intracellular proteins of feline immunodeficiency virus and their antigenic relationship with equine infectious anaemia virus proteins. Feline immunodeficiency virus (FIV) grown in cat lymphocyte and thymocyte cultures was labelled with L-[35S]methionine or [3H]glucosamine and virus-coded proteins were identified using immunoprecipitation. Polypeptides with apparent Mr values of 15K, 24K, 43K, 50K, 120K and 160K were detected. An additional polypeptide of 10K was detected by Western blot analysis. The two highest Mr species sometimes appeared as one band, of which only the 120K polypeptide was glycosylated. In the presence of tunicamycin gp120 was no longer detectable and a non-glycosylated precursor of 75K was found instead. ...
Immunogenicity and allergenicity of Culicoides imicola (Diptera: Ceratopogonidae) extracts. Summer seasonal recurrent dermatitis (SSRD) or "sweet itch" is a seasonally occurring allergic dermatitis of horses provoked by biting midges. The allergic skin reactions have been attributed to allergens present in various Culicoides species. C. imicola is the suspected etiological agent of SSRD in Israel. Whole body extracts of this midge induced hypersensitivity reactions upon injection into susceptible horses and in this study attempts were made to define components of C. imicola which have immunogenic and allergenic properties. Immunogenic potency was evaluated by raising antisera to whol...
Equine rabies immune globulin: a product with an undeserved poor reputation. Four hundred nineteen patients exposed to rabies in Thailand were treated with equine rabies immune globulin (ERIG) manufactured by Sclavo of Italy, a product also licensed in the United States of America. They were followed for a minimum of 1 month after ERIG injection and rabies vaccine administration. Adverse serum sickness-like reactions were noted in 15 patients (3.58%). These were clinically acceptable and only 1 of these patients required corticosteroid therapy and short term hospitalization for serum sickness. ERIG is approximately 1/10 of the cost of human rabies immune globulin (HRIG...
Comparison of heparan sulfate proteoglycans from equine and human glomerular basement membranes. 1. Proteoglycans extracted from human and equine glomerular basement membranes (GBM) were purified by ion-exchange chromatography and gel filtration. 2. The glycoconjugates had an apparent molecular mass of 200-400 kDa and consisted of 75% protein and 25% glycosaminoglycan. Glycosidase and HNO2 treatment and the amino sugar and sulfate composition of both proteoglycan preparations identified heparan sulfate (HS) as the predominant saccharide chain. 3. Hydrolysis with trifluoromethanesulfonic acid yielded comparable core proteins with molecular masses of ca 160 and 120 kDa. 4. The HS chains had...
[Heteroimmune hemolytic anemia associated with antilymphocyte globulin treatment in a patient with aplastic anemia]. A 24-year-old male patient with a severe aplastic anemia (SAA) was treated with equine-antilymphocyte globulin (ALG). As complication of this treatment he developed a severe heteroimmune hemolytic anemia mediated by anti-species pan-agglutinin antibodies present in ALG. In spite of the fact that ALG is absorbed with red-cell stroma and platelets to remove anti-erythrocyte and anti-platelet contaminating antibodies, often only partial absorption is achieved, and the remaining antibodies are passively acquired by the recipient. Neutropenia and especially thrombocytopenia are usual complications ...
[The development of the anti-phospholipase A2 antibody response in horses inoculated with venom for the production of polyvalent antisnake serum in Costa Rica]. The development of antibody response against phospholipase A2 activity of Bothrops asper venom was studied in a group of adult and healthy horses used in the production of the polyvalent antivenom at the Instituto Clodomiro Picado. Simultaneously, the general condition of the animals during the immunization schedule was also studied. There was a great individual variability in the immune response, although most of the horses studied reached the highest neutralizing titer after injection of doses of venom of 30 mg and 50 mg. On the other hand, in horses that had been previously immunized and we...
Antibodies to staphylococcal DNases in sera from different animal species, including humans. 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 ...
Antibody response to Ehrlichia risticii and antibody reactivity to the component antigens in horses with induced Potomac horse fever. The antibody response and the antibody reactivity to component antigens of Ehrlichia risticii were studied in horses with induced Potomac horse fever. These horses had no detectable antibodies to E. risticii in their preinoculation (PrI) sera by indirect fluorescent-antibody assay and enzyme-linked immunosorbent assay (ELISA). All the horses exhibited typical disease features following experimental infection and responded with specific antibodies, as measured by ELISA and indirect fluorescent-antibody assay. A primary antibody response was detected in 70% of the horses, while a secondary-type ...
Radioimmunoassay of inhibin in various mammals. A sensitive radioimmunoassay (RIA) for the determination of inhibin in peripheral plasma and tissue homogenates of different species has been developed using antisera to partially purified bovine follicular fluid (bFF) inhibin and 125I-labelled bFF 32 kDa inhibin. Antisera were produced by immunization of rabbits with partially purified bFF inhibin prepared by immunoaffinity chromatography. Increasing doses of a high titre antiserum could neutralize the suppressing effect of bFF, porcine follicular fluid and rat ovarian homogenate on FSH secretion from rat anterior pituitary cells in culture. ...
Evaluation of the opsonic capacity of core lipopolysaccharide antiserum of equine origin against smooth Escherichia coli 0111:B4, using macrophage chemiluminescence. A study was performed to determine whether equine antiserum to core lipopolysaccharide (LPS) would enhance phagocytosis of smooth gram-negative (GN) organisms by equine macrophages. Five healthy adult horses (group A) were immunized with a bacterin prepared from the J-5 mutant of Escherichia coli 0111:B4 and Salmonella minnesota R595 to produce antibodies to core LPS. Five horses (group B) served as nonimmunized controls and were given physiologic saline solution instead of the rough mutant bacterin. Serum antibody titers to core LPS and to smooth E coli 0111:B4 were determined by indirect ELI...
Surface antigens on equine sarcoid cells and normal dermal fibroblasts as assessed by xenogeneic antisera. To characterise the expression of surface antigens on equine sarcoid cells compared to normal equine fibroblasts, immune sera were produced in rabbits against transformed cells of a virus-containing sarcoid cell line (Mc-1) and normal dermal fibroblasts, respectively. The specificities of the sera were analysed by antibody-dependent cellular cytotoxicity against 51Cr-labelled target cells using human lymphocytes as effector cells. Anti-Mc-1 antiserum induced strong cytotoxicity against transformed cells of two sarcoid cell lines (Mc-1 and Bay Mc-1), whereas the cytotoxicity against transformed...
Isolation of enterotoxigenic Escherichia coli from a foal with diarrhea. Enterotoxigenic Escherichia coli was isolated from a 3-day-old foal with diarrhea. The isolate was distinguished from nonpathogenic E coli by determining the presence of pili and enterotoxin production. A standard slide agglutination test was performed, using pooled antisera that contained antibodies against K99 and F41 pilus antigens, K87 capsular antigen, and 0101 somatic antigen. Agglutination of the antisera occurred in the presence of the isolate. Piliation was verified by use of negative-contrast electron microscopy. Further, the isolate produced a heat-labile enterotoxin-like antigen th...
Complement fixing antibodies against arboviruses in horses at Lagos, Nigeria. Sixty-two sera horse collected from two stables at Lagos, Nigeria, were tested for complement fixing antibody to 8 arbovirus antigens; Chikungunya, Igbo-Ora, Yellow fever, Wesselsbron, West Nile, Potiskum, Uganda S and Rift Valley fever. Ten per cent of the horse sera examined contained CF antibody to one or more of the test antigens and indicated considerable arbovirus activity in the two stables. Reactions with flavivirus antigens were most common and the highest antibody titres were obtained with Wesselsbron and Yellow fever viruses. Eleven per cent of the sera tested reacted with alphaviru...
A previously reported polymorphic plasma protein of dogs and horses, identified as apolipoprotein A-IV. By using immunoblotting with antiserum specific to human plasma apolipoprotein A-IV (apoA-IV), a previously reported polymorphic plasma protein of dogs viz postalbumin-2 (Pa2) and one of horses viz serum protein 2 (SP2), were identified as apoA-IV of these species. This along with earlier published results implied that: (1) both dog and horse show a high degree of polymorphism at the APOA4 locus with three common alleles in each of the two species; and (2) apoA-IV phenotyping in these two species can be done by analysing plasma/serum samples by a simple method of two-dimensional electrophoresi...
Immunization of equines with phospholipase A2 protects against the lethal effects of Crotalus durissus terrificus venom. Equines (2 horses and 2 donkeys) immunized with whole Crotalus durissus terrificus venom or its phospholipase A2 component either presented an increased survival time determined 3 days after challenge or were totally resistant to a challenging lethal dose of 200 mg crude venom 270 days after the initial immunization or 90 days after the last booster injection. The resistance was demonstrable on the basis of a good correlation with antibody titers determined by the ELISA method but not with the flocculation and neutralization assays. Since phospholipase A2 is essentially nontoxic, it can be use...