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.
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...
Feeding patterns of Haemagogus janthinomys (Diptera: Culicidae) in different regions of Brazil. New data on the feeding patterns of Haemagogus (Haemagogus) janthinomys Dyar from different geographical regions of Brazil, by using the precipitin test as the bloodmeal-identifying tool, are presented. The following antisera were used: bird, dog, human, rodent, cattle, horse, and opossum. The origins of 287 bloodmeals were identified, whereas 33 specimens were negative to the antiserums tested. Among the reactive specimens, 174 (60.6%) fed on only one food source, of which 35.1% originated from birds, 19.5% from rodents, 12.6% from humans, 10.3% from cattle, 10.3% from opossums, 7.5% from dog...
Efficacy demonstration of tetanus vaccines by double antigen ELISA. This paper describes a double antigen ELISA (DAE) for rapid, specific and reliable assessment of the antitetanus immune status of horses and sheep. Compared with the indirect ELISA, the double antigen ELISA has the advantage of species-independent testing of sera. Thanks to its test design, it is more specific since the detected antibodies are forced to bind tetanus toxoid twice. In addition, it is very sensitive to tetanus antibodies, enabling the detection of low antibody titres, in range which is relevant for the assessment of the protective status (tetanus toxin neutralising antibodies). T...
LruA and LruB, novel lipoproteins of pathogenic Leptospira interrogans associated with equine recurrent uveitis. Recurrent uveitis as a sequela to Leptospira infection is the most common infectious cause of blindness and impaired vision of horses worldwide. Leptospiral proteins expressed during prolonged survival in the eyes of horses with lesions of chronic uveitis were identified by screening a phage library of Leptospira interrogans DNA fragments with eye fluids from uveitic horses. Inserts of reactive phages encoded several known leptospiral proteins and two novel putative lipoproteins, LruA and LruB. LruA was intrinsically labeled during incubation of L. interrogans in medium containing [14C]palmiti...
Glycoprotein G deletion mutants of equine herpesvirus 1 (EHV1; equine abortion virus) and EHV4 (equine rhinopneumonitis virus). Glycoprotein G (gG) deletion mutants of EHV1 and EHV4, designated EHV1DeltagG and EHV4DeltagG, were constructed. The growth characteristics of the EHV1DeltagG mutants were similar to the parent virus. All of the EHV4DeltagG mutants grew more slowly in cell culture and produced plaques of different morphology including smaller size. The yields of both gG deletion mutant viruses in cell culture were similar to the parent viruses. Sequencing of the genes flanking gG, Southern blot, PCR and western blot analyses of the mutant viruses demonstrated that the deletions were as expected, except for EHV...
Heterogeneous clearance of antithymocyte globulin after CD34+-selected allogeneic hematopoietic progenitor cell transplantation. Antithymocyte globulins (ATG) are purified, concentrated preparations of polyclonal immunoglobulin G from hyperimmune serum of horses or rabbits immunized with human thymus lymphocytes. Both the horse and the rabbit products induce immunosuppression as a result of lymphocyte depletion and immune modulation. The exact mechanism of action is unknown but may include T-cell clearance from the circulation and modulation of T-cell activation, homing, and cytotoxic activities. Both horse and rabbit ATG include multiple antibodies against T-cell surface antigens and have been used extensively in allog...
The improvement of the therapeutic anti-Lachesis muta serum production in horses. The main features associated with pit viper envenomations include the intense local lesions such as oedema, necrosis, acute renal failure and other effects. The severity of these reactions to snakebite depends on the degree of envenomation. Lachesis muta venom (LMV) has weak lethal activity, but due to the large amount often inoculated, the effects are extremely severe and demand anti-venom with a high neutralizing capacity. LMV had the lowest neutralizing antibody induction capacity in horses when compared with that of other venoms. For example, Bothrops anti-venom serum neutralizes 180 times...
Measurement of erythrocyte carbonic anhydrase isozymes (CA-I and CA-II) in racehorses and riding horses. Equine carbonic anhydrase isozymes (CA-I and CA-II) were purified from erythrocytes by several column chromatography. Polyclonal anti-CA-I and anti-CA-II sera were produced in rabbits. Sensitive competitive enzyme-linked immunosorbent assays (ELISA) were established to determine the developmental changes in CA-I and CA-II levels in equine erythrocytes. Concentrations of CA-I and CA-II in erythrocytes from 150 clinically normal thoroughbreds (123 racehorses and 27 riding horses) were determined by ELISA. Mean (+/- SD) concentrations of CA-I and CA-II in racehorses were 1.70 +/- 0.48 and 0.94 +/...
Systemic AL amyloidosis associated with multiple myeloma in a horse. AL amyloidosis is the most common type of systemic amyloidosis in humans, and it is frequently associated with multiple myeloma. But, AL amyloidosis is very rare in domestic animals. A 16-year-old Quarter horse gelding was diagnosed with systemic AL amyloidosis associated with multiple myeloma. Clinical problems were rapid weight loss, muscle atrophy, soft unformed stool, and ventral edema. Grossly, diffuse gastrointestinal hemorrhage, markedly thickened jejunal mucosa, and splenomegaly were present. Microscopically, diffuse severe amyloid deposits were present in the lamina propria of glandul...
Anticomplementary activity of equine whole IgG antivenoms: comparison of three fractionation protocols. Early adverse reactions occur in a number of patients treated with heterologous antivenoms and have been associated with anticomplementary activity (ACA). In order to reduce the ACA of equine whole IgG antivenoms produced by caprylic acid fractionation, three different fractionation protocols were compared: (a) routine caprylic acid fractionation; (b) caprylic acid fractionation followed by beta-propiolactone treatment; and (c) caprylic acid fractionation followed by ion-exchange chromatography using a quaternary ammonium membrane. The three protocols yielded products with similar physicochemi...
Endocrine-paracrine cells of the male urogenital apparatus: a comparative histochemical and immunohistochemical study in some domestic ungulates. Specimens of testis, excurrent duct including the male accessory glands and urethra, were studied in boars, bulls, horses and donkeys, in order to localize endocrine/paracrine cells. Silver impregnation methods were used to test the argentaffinity and/or argyrophilia of cells. Immunoreactivities to chromogranin A, 5-hydroxytryptamine, somatostatin, [met]- and [leu]- enkephalins, gastrin-releasing peptide, calcitonin gene-related peptide, neuropeptide Y, substance P, vasoactive intestinal peptide, beta-endorphin antisera were tested by a streptavidin-biotin method. In the testis, epididymis, du...
Phage display of the Equine arteritis virus nsp1 ZF domain and examination of its metal interactions. A putative zinc finger (ZF) domain in the Equine arteritis virus (EAV) nsp1 protein was described recently to be required for viral transcription. The nsp1 ZF (50 aa) was expressed on the surface of M13KE gIII phage, fused to the N terminus of the phage pIII protein. To evaluate the functionality of the ZF domain, a binding assay was developed, based on the use of immobilized Ni(2+) ions (Ni-NTA). Phages displaying ZF bound significantly better to Ni-NTA than did phages displaying negative-control peptides, which also contained metal-coordinating residues. Also, binding of ZF-displaying phages...
Genetic characterization of equine arteritis virus during persistent infection of stallions. Equine arteritis virus (EAV) causes a persistent infection of the reproductive tract of carrier stallions. The authors determined the complete genome sequences of viruses (CW96 and CW01) that were present 5 years apart in the semen of a carrier stallion (CW). The CW96 and CW01 viruses respectively had only 85.6 % and 85.7 % nucleotide identity to the published sequence of EAV (EAV030). The CW96 and CW01 viruses had two 1 nt insertions and a single 1 nt deletion in the leader sequence, and a 3 nt coding insertion in ORF1a; thus their genomes included 12 708 nt as compared to the 12 704 nt in EA...
First case report of Sarcocystis neurona-induced equine protozoal myeloencephalitis in Japan. Equine protozoal myeloencephalitis developed in a three-year-old male Thoroughbred racehorse imported from the United States. The animal showed astasia five days after the onset of ataxia. Histopathologically, focal nonpurulent myelitis accompanied by hemorrhage and perivascular infiltration was observed in the fourth and fifth cervical spinal cord. Immunohistochemically, shizonts were occasionally observed and were positive for anti-Sarcocystis neurona (S. neurona) antiserum. S. neurona-specific antibodies were detected in the serum and cerebrospinal fluid by Western blot. This is the first e...
Equine trypsin: purification and development of a radio-immunoassay. Shock is accompanied by generalised splanchnic hypoperfusion, and splanchnic organs like the pancreas can be damaged, as shown in animal experimental models and in humans, by the presence of high plasma concentrations of trypsin and other pancreatic enzymes. In order to design a radioimmunoassay technique (RIA) for the measurement of equine trypsin-like immunoreactivity (TLI) in biological fluids, trypsin was purified (with purity > or = 96%) from the equine pancreas by extraction in an acid medium, ammonium sulfate precipitations, gel filtration chromatography and, after activation of tryp...
Mapping epitopes in equine rhinitis A virus VP1 recognized by antibodies elicited in response to infection of the natural host. Equine rhinitis A virus (ERAV) is an important respiratory pathogen of horses and is of additional interest because of its close relationship and common classification with foot-and-mouth disease virus (FMDV). As is the case with FMDV, the VP1 capsid protein of ERAV has been shown to be a target of neutralizing antibodies. In FMDV VP1, such antibodies commonly recognize linear epitopes present in the betaG-betaH loop region. To map linear B cell epitopes in ERAV VP1, overlapping fragments spanning its length were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins....
IgE ELISA using antisera derived from epsilon chain antigenic peptides detects allergen-specific IgE in allergic horses. Equine disease with an allergic etiology is common. Environmental antigens most often implicated as allergens in horses include molds, dusty hay, grass pollen, hay dust mites, and insect saliva. Although intradermal testing with allergen is a useful diagnostic tool for some species, skin testing frequently produces false positive results in horses. Allergen deprivation as a diagnostic tool is often impossible and at best it is ineffective at diagnosing the specific allergic reactivity. Synthesis of IgE after exposure to allergen is the instigator of the allergic process. While IgE exerts its e...
Collaborative study for the establishment of two European Pharmacopoeia Biological Reference Preparations for serological potency testing of tetanus vaccines for veterinary use. The European Directorate for the Quality of Medicines (EDQM) has organised an international collaborative study, divided into two phases, aimed at producing and establishing two suitable reference sera for serological potency testing of tetanus vaccines for veterinary use for batch consistency demonstration. In phase I pools of sera were produced by immunising guinea pigs and rabbits with tetanus toxoid using the immunisation schedule prescribed by the European Pharmacopoeia (Ph. Eur.) for potency testing of tenanus vaccines for veterinary use. Following aliquoting and freeze-drying, character...
Collaborative study for the establishment of a European Pharmacopoeia Biological Reference Preparation for Clostridia antiserum for serological potency testing of veterinary clostridial vaccines. The European Directorate for the Quality of Medicines (EDQM) has organised an international collaborative study, divided in two phases, aimed at producing and establishing a suitable reference serum for serological potency testing of clostridial vaccines for batch consistency demonstration. In phase 1 a series of pools produced from sera provided by each manufacturer and raised against the broadest range of antigens possible were blended to obtain TN titres which were representative of the range normally elicited by the vaccines under test. Detailed statistical analysis of the data was not pos...
The serologic response of horses to equine arteritis virus as determined by competitive enzyme-linked immunosorbent assays (c-ELISAs) to structural and non-structural viral proteins. In an effort to further characterize the humoral immune response of horses to equine arteritis virus (EAV), direct and competitive enzyme-linked immunosorbent assays (c-ELISAs) were developed using monoclonal and polyclonal anti-sera to structural (G(L), N and M) and non-structural (nsp1) viral proteins. A nsp1-specific monoclonal antibody was produced to facilitate development of a c-ELISA to this protein. Data obtained using the various c-ELISAs confirm that the M protein is a major target of the antibody response of horses to EAV. However, none of the c-ELISAs that were developed were as se...
Inhibition of collagenase breakdown of equine corneas by tetanus antitoxin, equine serum and acetylcysteine. To determine whether tetanus antitoxin, equine serum, and acetylcysteine, which are currently used in the treatment of equine corneal ulcer, inhibit the digestion of equine corneal collagen when exposed to collagenase in vitro. Methods: Corneas from 40 adult horses. Methods: Sections of equine corneas were incubated with saline, a solution of bacterial collagenase in saline, bacterial collagenase in saline plus equine tetanus antitoxin, bacterial collagenase in saline plus equine serum, or bacterial collagenase in saline plus acetylcysteine. Each one of the collagenase inhibitors was tested at...
The impact of a low dose, low volume, multi-site immunization on the production of therapeutic antivenoms in Thailand. Therapeutic antivenom against snakes was first produced by Albert Calmette in 1894. Since then antivenoms have saved the life of countless snakebite victims. However, there are still many problems associated with antivenom production, for example variable percentage of responder horses, low neutralizing potency of antivenom, the large amount of snake venom needed for immunization and the difficulties encountered in producing potent polyvalent antivenoms. These problems have led to shortage and high cost of antivenom and, in some cases, failure of treatment. In 1997, a new immunization protocol...
Equid herpesvirus 1 infection of endothelial cells requires activation of putative adhesion molecules: an in vitro model. Antisera to activated equine endothelial cells, which detected surface molecules of 116 kD, 97 kD, 42 kD and 38 kD, were made to investigate the role of endothelial adhesion molecules in equid herpes virus 1 infection. These putative adhesion molecules could be induced by 17-beta oestradiol, chorionic gonadotrophin, or IL-2, as well as by LPS and PWM. In an in vitro flow system, using equine veins or arteries, equid herpesvirus 1 in leucocytes was only transferred to infect endothelial cells if both leucocytes and endothelial cells expressed these surface molecules. Blocking of the membrane mo...
Inactivation of West-Nile virus during peptic cleavage of horse plasma IgG. Peptic cleavage of horse plasma IgG is a common procedure for the preparation of F(ab)(2) products for human use, such as antivenin and antitoxin. The removal of the Fc fragment from the IgG molecule by enzymatic cleavage at low pH, ensures fewer side-effects of the F(ab)(2) product for passive immunotherapy compared with the whole IgG molecule. Since the starting material may be contaminated by zoonotic horse viruses, it is necessary to demonstrate the removal or inactivation of possible viral contaminants. Guidelines for performing such studies were published by the Commission for Plasma-Der...
Conservation of recognition of antibody and T-cell-defined alloantigens between species of equids. Serological and cellular assays and molecular techniques were used to define features of the major histocompatibility complex (MHC) of the donkey With this information in hand, immune recognition of MHC determinants within and between donkeys and horses was compared. An antibody-mediated, complement-dependent, microcytotoxicity assay using a variety of antisera to donkey histocompatibility antigens, including those induced as a result of intraspecies or interspecies pregnancy in horse mares and jenny donkeys, delineated five donkey leukocyte antigen (DoLA) specificities. Antisera raised across...
Importance of M-protein C terminus as substrate antigen for serodetection of equine arteritis virus infection. 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...
Functional expression and membrane fusion tropism of the envelope glycoproteins of Hendra virus. Hendra virus (HeV) is an emerging paramyxovirus first isolated from cases of severe respiratory disease that fatally affected both horses and humans. Understanding the mechanisms of host cell infection and cross-species transmission is an important step in addressing the risk posed by such emerging pathogens. We have initiated studies to characterize the biological properties of the HeV envelope glycoproteins. Recombinant vaccinia viruses encoding the HeV F and G open reading frames were generated and glycoprotein expression was verified by metabolic labeling and detection using specific antis...