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.
[Comparative efficacy of homologous and heterologous biologicals against diphtheria]. Comparative assessment of efficacy of homologous and heterologous diphtheria antitoxins on the example of diphtheria intoxication. Methods: Homologous hyperimmune sera were obtained through immunization of rabbits and guinea-pigs with diphtheria toxoid according to schedule. Immune rabbit sera contained 70 - 100 IU/mL of antitoxin antibodies and guinea-pig sera contained 60 - 80 IU/mL. Equine diphtheria antitoxin was used as a heterologous one. Measurement of antitoxin level using experimental animals is based on quantitative assessment of ability of studied sera to neutralize specific dermone...
Evaluation of neutralization patterns of the five unique Argentine equine arteritis virus field strains reported. Equine viral arteritis (EVA) is a contagious viral disease that frequently causes mild or subclinical infections in adult horses. Only one EAV serotype has been described. However, there are differences in antigenicity, pathogenicity and neutralization characteristics of virus field strains. The interaction of two viral proteins, GP5 and M, is critical for infectivity and amino acid changes in the GP5 sequences have an effect on the neutralizing phenotype, regardless the effects of other viral proteins. The objective of the present study was to evaluate the neutralization phenotypes of the 5 u...
Pharmacokinetics in rabbits and anti-sphingomyelinase D neutralizing power of Fab, F(ab’)(2), IgG and IgG(T) fragments from hyper immune equine plasma. We describe the separation of whole IgG, IgG(T)-less IgG (called here merely IgG) and IgG(T) and the production of Fab and F(ab')(2) fragments. We studied the pharmacokinetics of these immunoglobulins and fragments in rabbits. Both, the isotypes and the whole IgG fragments were purified and/or produced from the same plasma lot from horses hyper immunized against sphingomyelinase D to produce anti-Loxosceles antivenom. The sphingomyelinase D neutralizing ability of the isotypes and their fragments was measured. Fab and F(ab')(2) PK was well described by a tri-exponential kinetics. IgG and IgG(T...
[Two horses with neurological symptoms: could this be equine botulism?]. Symptoms, diagnosis and therapy of equine botulism are discussed by the presentation of two detailed reports of horses with neurological symptoms and the results of laboratory investigations over the period 2003-2008 in the Netherlands. In addition a brief summary of the available literature is presented. Prevailing symptoms of botulism in horses include paralysis of the tongue, salvation, dysphagia and paresis and paralysis of the skeletal muscles, as well as signs of colic. Symptoms and prognosis vary with the amount of botulinum neurotoxin (BoNT) involved. For early clinical diagnosis of bo...
Identification and characterisation of an immunodiagnostic marker for cyathostomin developing stage larvae. Parasitic nematodes of the group Cyathostominae are an important cause of disease in horses. This group consists of approximately 50 species, all of which have similar life cycles that involve encystment of larval stages in the large intestinal wall. Encysted larvae can persist for months to years and, occasionally, large numbers can accumulate and emerge synchronously to cause severe pathology, resulting in diarrhoea, weight loss, colic and/or oedema. This syndrome, known as larval cyathostominosis, can be fatal in up to 50% of cases, despite treatment. There is no diagnostic method that enab...
Evidence for a novel vasospastic transmitter system, neuromedin U, in the equine digital circulation. The brain-gut peptide neuromedin U (NMU) is a ligand for the G-protein-coupled receptors, NMU1 and NMU2. In humans, an extended form of this peptide, NMU-25, and the structurally related peptide, neuromedin S (NMS), both produce potent vasoconstriction in isolated blood vessels. The aim of this study was to determine whether NMU fulfilled criteria for controlling vasoreactivity in the equine digital circulation. NMU receptors were characterised in the equine digital artery and vein based on the pharmacological criteria of specific, saturable and high affinity binding. Immunoreactive peptide wa...
Stability of equine IgG antivenoms obtained by caprylic acid precipitation: towards a liquid formulation stable at tropical room temperature. Liquid formulations of antivenom require a cold chain for their distribution and storage, especially in tropical countries characterized by high temperature and humidity (climatic zone IV). Since cold chain is often deficient in many regions, there is a need to develop novel formulations of liquid antivenoms of higher stability at room temperatures. The effect of addition of the polyols mannitol and sorbitol on the thermal stability of caprylic acid-fractionated equine whole IgG antivenoms was assessed in preparations having different concentrations of protein and phenol. Results evidenced tha...
Paraspecific neutralization of the venom of African species of cobra by an equine antiserum against Naja melanoleuca: a comparative study. Venoms of snakes belonging to the same Genera tend to share biochemical, toxinological and antigenic characteristics. Accordingly, paraspecific neutralization of venom lethality by experimental antisera and commercial antivenoms has been reported. We studied the spectrum of neutralization of lethality of an experimental monovalent equine antiserum against the strongly neurotoxic African forest cobra (Naja melanoleuca) when tested against venoms of most species of African Naja, both neuro and cytotoxic as described by some authors. We report a comparison of the median lethal doses (LD50) of the...
Rational design of synthetic peptides to generate antibodies that recognize in situ CD11c(+) putative dendritic cells in horse lymph nodes. A three-dimensional model of the alphaX I-domain of the horse integrin CD11c from dendritic cells provided information for selecting two segments of the primary structure for peptide synthesis. Peptide 1 contains 20 amino acids and peptide 2 has 17 amino acid residues. The first spans from position Thr229 to Arg248 of an alpha-helix segment of the structure, whereas peptide 2 goes from Asp158 to Phe174 and corresponds to an exposed segment of the loop considered to be the metal ion-dependent adhesion site. Murine polyclonal antisera against both peptides were generated and assayed in periphera...
Standardization and validation of Vero cell assay for potency estimation of diphtheria antitoxin serum. Diphtheria toxin has the capacity to block protein synthesis in cultured mammalian cells, and thus causing cell death. This capacity of diphtheria toxin was utilized for in-vitro neutralization test to determine antibody titer, using Vero cells, which have been found to be susceptible to diphtheria toxin. In the present study, a Vero cell assay was standardized and validated for potency estimation of diphtheria antitoxin serum (DATS). The results obtained by Vero cell assay were compared with in-vivo biological assay. High degree of correlation (+0.98) was found between in-vivo biological assa...
Assessment of the anti-Naja haje antibodies elicited in a low dose multi-site immunization protocol. The horse antibodies to Naja haje (Nh) elicited in a low dose multi-site immunization protocol were investigated from binding perspective in the context of antivenom maturation. We found that, this protocol evoked the production of lethality neutralizing avid antibodies in the first round of immunization which increases over the successive immunization rounds. The changes and the relative changes in the antibody parameters of each horse were taken as a measure for the efficacy of its immune system to respond by gaining and magnifying the antivenom parameters. While the avidity increases over t...
Comparison of Montanide adjuvants, IMS 3012 (Nanoparticle), ISA 206 and ISA 35 (Emulsion based) along with incomplete Freund’s adjuvant for hyperimmunization of equines used for production of polyvalent snake antivenom. The use of adjuvant is of fundamental importance in vaccines formulations and antisera production. Currently selection and use of adjuvant systems in snake antivenom preparation has become a major issue in terms of animal welfare as well as economics. In order to minimize disadvantages associated with traditionally used Freund's adjuvant (FA) in equines and to produce potent polyvalent antivenom against four Indian snake venoms in minimum possible period, a comparison was made between various commercially available non-emulsion/emulsion based adjuvants like IMS 3012, ISA 206 and ISA 35 with In...
Effects of bites by the European adder (Vipera berus) in seven Swedish horses. The effects on seven horses of bites by the European adder (Vipera berus) are described and compared with previously available information. The clinical signs varied from local swelling and mild systemic signs to severe systemic signs, including systemic inflammatory response, severe tissue necrosis, ventricular tachycardia and dysphagia. Two of the horses were treated with 'Zagreb' antiserum, and three that were not treated with antiserum were euthanased owing to complications related to the bites.
Collaborative study for the establishment of a candidate equine influenza subtype 2 American-like strain A/EQ/South Africa/4/03 – horse antiserum biological reference preparation. In 2004, the Office International des Epizooties (OIE) Expert Surveillance Panel on equine influenza recommended that the American lineage component (H3N8) of equine influenza vaccines (A/eq/Newmarket/1/93-like) be updated to an A/eq/South Africa/4/03-like virus. As a consequence the common European Pharmacopoeia (Ph. Eur.) - OIE reference for equine influenza subtype 2 American-like antiserum had to be complemented by an antiserum raised in horses against an A/eq/South Africa/4/03 strain. An international collaborative study run by the European Directorate for the Quality of Medicines (EDQM) ...
Evaluation of a rapid agglutination method for detection of equine red cell surface antigens (Ca and Aa) as part of pretransfusion testing. Blood typing before transfusion minimizes the risk of transfusion reactions and prevents immunization of the recipient against incompatible RBC antigens. The major RBC antigens that warrant identification before packed RBC or whole blood transfusions in horses are Ca and Aa. Standard blood-typing protocols are time-consuming (2.5-3.0 hours) and impractical in emergency settings. Objective: The purpose of this study was to determine whether equine RBCs could be typed for Ca and Aa antigens using sera from horses with RBC antibodies in a modified rapid (15 minute) blood-typing protocol. Methods:...
Characterization of immunogenic proteins of Trypanosoma evansi isolated from three different Indian hosts using hyperimmune sera and immune sera. The western blot analysis for identification of immunogenic proteins in whole cell lysate (WCL) antigens (Ags) prepared from the Trypanosoma evansi of buffalo, horse and cattle origins using hyperimmune sera (HIS) showed 11 immunogenic proteins and naturally T. evansi infected immune sera (IS) of horse detected 19 immunogenic proteins. HIS and IS of horse recognized five common immunogenic proteins of relative molecular weight (M(r)) ranges 61-64, 44-47, 33-34, 25-26 and 14-16 kilo Dalton (kDa). HIS rose against WCL Ags of T. evansi of buffalo origin and immune sera of horse cross reacted with...
Histomonas meleagridis (Parabasala, Trichomonadea, Monocercomonadidae): presence of natural agglutinins in horse serum. Cultured Histomonas meleagridis cells were readily agglutinated in vitro by horse serum at concentrations as low as 5%, although clumping was more rapid and prominent at 15% or higher. For observation of clumping, the cultured organisms were washed twice in Hanks balanced solution (HBSS) by centrifugation (1,000 x g for 15 min) and filtered through glass wool. The test sera were added and the mixture incubated in a Petri plate or 24-well culture plates at r.t. for 15-30 min. Formation of clumps was time- and concentration-dependent. Gentle agitation hindered agglutination at low serum concentr...
IdeE reduces the bactericidal activity of equine neutrophils for Streptococcus equi. Streptococcus equi (S. equi) causes equine strangles, a highly contagious and widespread purulent lymphadenitis of the head and neck. Highly resistant to phagocytosis, it produces long extracellular chains in affected lymph nodes. In a screen of clones reactive with convalescent serum from a gene library of S. equi CF32 we identified IdeE, an IgG-endopeptidase and homologue of the leucocyte receptor Mac-1 (CD11b). IdeE is expressed during S. equi infection eliciting both serum and mucosal antibody responses which persisted at significant levels in serum for over 200 days. Release from S. equi ...
Immunohistochemical localization of vascular endothelial growth factor (VEGF) and its two receptors (Flt-I and KDR) in the endometrium and placenta of the mare during the oestrous cycle and pregnancy. Polyclonal antisera for vascular endothelial growth factor (VEGF) and its two main receptor molecules, VEGF-I (Flt) and VEGF-II (KDR), were used in a conventional immunocytochemical staining method to localize these angiogenic ligand molecules in the endometrium and placenta of the mare during the oestrous cycle and pregnancy. The anti-VEGF and anti-Flt sera both labelled the lumenal and glandular epithelia of the endometrium throughout the oestrous cycle and both the invasive trophoblast cells of the endometrial cups and the non-invasive trophoblast of the allantochorion in pregnancy. The ant...
Purification of equine IgG using membrane based enhanced hybrid bioseparation technique: a potential method for manufacturing hyperimmune antibody. Hyperimmune equine IgG is widely used as antivenom and anti-rabies agents. This article discusses a membrane based enhanced hybrid bioseparation technique for efficient and scalable purification of equine immunoglobulin G (IgG) from horse serum. This technique is an improved version of a standard hybrid bioseparation technique developed within our group earlier for fractionation of human plasma proteins (Ghosh. 2004. J Membr Sci 237: 109-117). In the presence of a high antichaotropic salt concentration, equine IgG is selectively and reversibly captured within a stirred cell membrane module fro...
On the presence of antibodies against bovine, equine and poultry immunoglobulins in human IgG preparations, and its implications on antivenom production. Specific immunoassays were developed to detect anti-horse, anti-chicken and anti-bovine immunoglobulins in human IgG preparations. Three samples of 5% human IgG for intravenous use ("Inmunoglobulina G Endovenosa al 5%"(trade mark), Quimbiotec CA), were studied. All samples were produced from pools of >2500 plasma units from different donors. One sample was produced from an only Venezuelan plasma pool (2660 donors) and the other two were produced from a 1:1 blend of Venezuelan and Canadian plasma pools. The amounts of human IgG detected were 0.017 (0.015,0.020) mg/ml (n=18) against horse IgG...
Immunolocalization of steroidogenic enzymes in equine fetal adrenal glands during mid-late gestation. To elucidate the relationship between steroidogenic hormones and developing adrenal glands, we investigated the immunolocalization of steroidogenic enzymes in equine fetal adrenal glands during mid-late gestation. Fetal adrenal glands were obtained from three horses at 217, 225 and 235 days of gestation. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17) and huma...
Expression and nephron segment-specific distribution of major renal aquaporins (AQP1-4) in Equus caballus, the domestic horse. Aquaporins (AQPs) play fundamental roles in water and osmolyte homeostasis by facilitating water and small solute movement across plasma membranes of epithelial, endothelial, and other tissues. AQP proteins are abundantly expressed in the mammalian kidney, where they have been shown to play essential roles in fluid balance and urine concentration. Thus far, the majority of studies on renal AQPs have been carried out in laboratory rodents and sheep; no data have been published on the expression of AQPs in kidneys of equines or other large mammals. The aim of this comparative study was to determ...
Comparative serological diagnosis of toxoplasmosis in horses using locally isolated Toxoplasma gondii. A total of 420 serum samples collected from horses of different ages, sexes and breeds, located at some horse farms in Egypt, were used for serological studies. A crude antigen of the locally isolated Toxoplasma gondii tachyzoites from horse tissues (LA) was used for the detection of T. gondii antibodies in horses. It showed good diagnostic efficiency (38.1%) by Enzyme Linked Immunosorbent Assay (ELISA). To increase this efficiency, an affinity purification process was performed. Two fractions were obtained from LA by CNBr-Sepharose 4B affinity column chromatography named; unbound (LAunb) and ...
Equine botulism and acute pasture myodystrophy: new soil-borne emerging diseases in Switzerland? In Switzerland, the incidence of equine botulism and acute pasture myodystrophy have remarkably increased in the last five years. Equine fodder-borne botulism in Europe is most likely caused by Clostridium botulinum types C and D that produce the toxins BoNT/C and BoNT/D. Horses showing signs suggestive of botulism (muscle weakness and tremors, reduced tongue tone, slow chewing, salivation and difficulties swallowing, drooping eyelids, mydriasis), especially patients that have fed on suspect fodder (mostly haylage), must be treated with anti-serum as soon as possible. They also need intensive ...
Fine and domain-level epitope mapping of botulinum neurotoxin type A neutralizing antibodies by yeast surface display. Botulinum neurotoxin (BoNT), the most poisonous substance known, causes naturally occurring human disease (botulism) and is one of the top six biothreat agents. Botulism is treated with polyclonal antibodies produced in horses that are associated with a high incidence of systemic reactions. Human monoclonal antibodies (mAbs) are under development as a safer therapy. Identifying neutralizing epitopes on BoNTs is an important step in generating neutralizing mAbs, and has implications for vaccine development. Here, we show that the three domains of BoNT serotype A (BoNT/A) can be displayed on the...
Presence of natural anti-Galalpha1-4GalNAcbeta1-3Gal (anti-NOR) antibodies in animal sera. Rare polyagglutinable NOR erythrocytes contain unusual globoside extention products terminating with a Galalpha1-4GalNAcbeta1-3Gal- unit. This trisaccharide epitope is recognized by recently characterized antibodies naturally occurring in most human sera (Duk et al., Glycobiology, 15, 109, 2005). These antibodies represent two major types of fine specificity. All these antibodies are most strongly inhibited by Galalpha1-4GalNAcbeta1-3Gal (NOR-tri), and weakly by Galalpha1-4Gal. However, the type 1 antibodies are strongly inhibited by Galalpha1-4Galbeta1-3Gal-R and weakly by Galalpha1-4GalNAc, ...
Horse embryonic stem cell lines from the proliferation of inner cell mass cells. Inner cell mass (ICM) cells were isolated immunosurgically from day 7-8 horse blastocysts and, after proliferation in vitro for 15-28 passages, three lines of cells were confirmed to be embryonic stem (ES) cells by their continued expression of alkaline phosphatase activity and their ability to bind antisera specific for the recognized stem cell markers, SSEA-1, TRA-1-60, TRA-1-81, and the key embryonic gene Oct-4. When maintained under feeder cell-free conditions in vitro, the three lines of cells differentiated into cells of ectodermal, endodermal, and mesodermal lineages. However, they did ...
Immunohistochemical identification and pathologic findings in natural cases of equine abortion caused by leptospiral infection. The aim of this study was to examine the utility of immunohistochemistry (IHC) in the diagnosis of leptospiral equine abortion and to compare IHC to silver staining and serology of the aborted mares. Ninety-six fetuses from 57 farms were examined using all 3 diagnostic techniques, revealing evidence of leptospiral infection in 3 fetuses (3.1%) from 3 (5.3%) different farms. A new finding in 1 of these confirmed cases of leptospiral abortion was the presence of macroscopic pinpoint grayish-white nodules that had a histologic correlate of hepatic necrosis; other histologic findings were consiste...
Passive immunotherapy for influenza A H5N1 virus infection with equine hyperimmune globulin F(ab’)2 in mice. Avian influenza virus H5N1 has demonstrated considerable pandemic potential. Currently, no effective vaccines for H5N1 infection are available, so passive immunotherapy may be an alternative strategy. To investigate the possible therapeutic effect of antibody against highly pathogenic H5N1 virus on a mammal host, we prepared specific equine anti-H5N1 IgGs from horses vaccinated with inactivated H5N1 virus, and then obtained the F(ab')2 fragments by pepsin digestion of IgGs. Methods: The horses were vaccinated with inactivated H5N1 vaccine to prepare anti-H5N1 IgGs. The F(ab')2 fragments were p...