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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.
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...
Antibody and cytokine-associated immune responses to S. equi antigens entrapped in PLA nanospheres. Strangles is an infectious disease caused by Streptococcus equi subspecies equi that affects the upper respiratory tract of the Equidae. The control of this disease seems to be dependent on its earlier detection and prevention, but prolonged animal protection without development of strong and severe side effects has not yet been achieved. Convalescent horses exhibit a protective immune response, mainly against SeM (58 kDa), an antiphagocytic and opsonogenic S. equi M-like protein, known as the major protective antigen against strangles. Purified recombinant SeM and S. equi protein extract-entr...
Immunisation of the equine uterus against Streptococcus equi subspecies zooepidemicus using an intranasal attenuated Salmonella vector. Attenuated Salmonella enterica serovar Typhimurium MGN707, expressing the SzP protective protein of the MB9 serovar of Streptococcus equi subspecies zooepidemicus (SzP-MB9) was tested for its safety and efficacy as a nebulised intranasal vaccine against streptococcal uterine infections in mares. In a preliminary study, vaccinated mares (n=5) displayed serum, nasal and uterine responses (P<0.05) to S. Typhimurium lipopolysaccharide (St-LPS). Subsequently, vaccinated mares (expressor group, n=7), but not mares vaccinated with the vector only (control group, n=7), displayed significant increas...
Latherin: a surfactant protein of horse sweat and saliva. Horses are unusual in producing protein-rich sweat for thermoregulation, a major component of which is latherin, a highly surface-active, non-glycosylated protein. The amino acid sequence of latherin, determined from cDNA analysis, is highly conserved across four geographically dispersed equid species (horse, zebra, onager, ass), and is similar to a family of proteins only found previously in the oral cavity and associated tissues of mammals. Latherin produces a significant reduction in water surface tension at low concentrations (< or = 1 mg ml(-1)), and therefore probably acts as a wettin...
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...
Localization of the equine IgG-binding domain in the fibrinogen-binding protein (FgBP) of Streptococcus equi subsp. equi. Fibrinogen-binding protein (FgBP, also termed SeM) is a cell-wall-associated anti-phagocytic M-like protein of the equine pathogen Streptococcus equi subsp. equi, and binds fibrinogen (Fg) and IgG. FgBP binds Fg avidly through residues located at the extreme N terminus of the molecule, whereas the IgG-binding site is more centrally located between the A and B repeats. FgBP binds equine IgG4 and IgG7 subclasses through interaction with the CH2-CH3 interdomain region of IgG-Fc, and possesses overlapping Fc-binding sites with protein A and protein G. In this study, FgBP truncates containing defin...
Expression of prostate glycoconjugates in the stallion and castrated horse. This work was undertaken to determine the glycoconjugates secreted by the epithelium of the prostate in the intact stallion and castrated horse using lectin histochemical procedures in conjunction with enzymatic digestion and deglycosylation treatments. Additionally, anti-5 and 13-16-cytokeratin antibodies were used to localize epithelial basal cells. In the stallion, lectin histochemistry showed the following sugar residues in the Golgi zone of the glandular cells: α-Glu/Man, α-Fuc and β-Gal included in both O- and N-linked oligosaccharides as well as β-GalNAc, GlcNAc and α-Gal, which be...
Prevalence and diagnosis of Babesia and Theileria infections in horses in Italy: a preliminary study. Babesia caballi and Theileria equi are the causative agents of equine piroplasmosis. In this preliminary epidemiological study, 412 horses reared in central and northern Italy were sampled and three diagnostic methods compared, namely, the microscopy, the indirect fluorescent antibody test (IFAT) and a PCR. Possible risk factors (such as area, season, breed, activity, sex, age, and grazing) associated with serological positivity were evaluated. A seroprevalence of 68.4% was found: 12.4% of the animals had anti-T. equi antibodies, 17.9% anti-B. caballi antibodies and 38.1% had antibodies agains...
Linear B-cell epitope mapping using enzyme-linked immunosorbent assay for libraries of overlapping synthetic peptides. The aim of this chapter is to provide a strategy for mapping linear antibody epitopes of protein antigens in order to discover candidates for vaccines or diagnostic tests. A set of overlapping peptides was designed and synthesised based upon a known amino acid sequence of the target protein, virulence-associated protein A (VapA) of the bacterium Rhodococcus equi, an important pulmonary pathogen in foals.The peptides were biotinylated and used in an ELISA to screen immune sera from foals. These biotinylated peptides were coated directly onto micro titre plates that had been pre-coated with Neut...
Intrinsic innervation of the horse ileum. This paper describes the morphology and distribution of the enteric nervous system (ENS) cells and fibres immunoreactive for choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), substance P (SP), calcitonin gene-related peptide (CGRP), NF200kDa (NF200), and S100 protein. The percentages of subclasses of enteric neurons in the total neuronal population were investigated by the use of anti-PGP 9.5 or anti-NSE antibodies. ChAT-IR myenteric plexus (MP) and submucosal plexus (SMP) neurons were 66+/-7% and 74+/-15%, respectively, whereas those cells expressing nNOS-IR were 38+/-7...
A perspective on Theileria equi infections in donkeys. The donkey population has remained unchanged in the last two decades despite a decrease in the overall population of equids, emphasizing the usefulness of the donkey as a draught and pack animal. Piroplasmosis in donkeys, caused by Theileria equi and Babesia caballi, has been recognized as a serious problem of major economic importance as the affected animals manifest decreased working capacity, loss of appetite, etc. In tropical countries, T. equi infections are more wide-spread and pathogenic than those caused by B. caballi. Donkeys usually remain asymptomatic carriers with positive antibody...
Experimental infection of potential reservoir hosts with Venezuelan equine encephalitis virus, Mexico. In 1993, an outbreak of encephalitis among 125 affected equids in coastal Chiapas, Mexico, resulted in a 50% case-fatality rate. The outbreak was attributed to Venezuelan equine encephalitis virus (VEEV) subtype IE, not previously associated with equine disease and death. To better understand the ecology of this VEEV strain in Chiapas, we experimentally infected 5 species of wild rodents and evaluated their competence as reservoir and amplifying hosts. Rodents from 1 species (Baiomys musculus) showed signs of disease and died by day 8 postinoculation. Rodents from the 4 other species (Liomys s...
Sandwich ELISA system for cartilage oligomeric matrix protein in equine synovial fluid and serum. Measurement of cartilage oligomeric matrix protein (COMP) in serum has potential for diagnosis of equine osteoarthritis (OA), but clinical use is currently limited by the lack of specificity of an inhibition ELISA as well as by baseline increases due to exercise. Improved methods for ELISA with increased antigen specificity and sensitivity are therefore required for reliable measurement. Objective: Measurement of the serum level of COMP by sandwich ELISA allows identification of horses with OA. Methods: New monoclonal antibodies (mAbs) were elicited against equine cartilage COMP, their epitope...
Influenza A viruses with truncated NS1 as modified live virus vaccines: pilot studies of safety and efficacy in horses. Three previously described NS1 mutant equine influenza viruses encoding carboxy-terminally truncated NS1 proteins are impaired in their ability to inhibit type I IFN production in vitro and are replication attenuated, and thus are candidates for use as a modified live influenza virus vaccine in the horse. Objective: One or more of these mutant viruses is safe when administered to horses, and recipient horses when challenged with wild-type influenza have reduced physiological and virological correlates of disease. Methods: Vaccination and challenge studies were done in horses, with measurement ...
Aggregation-associated loss of antigenicity observed for denatured virion protein 1 of Equine rhinitis A virus in an enzyme-linked immunosorbent assay. Equine rhinitis A virus (ERAV) is a picornavirus which causes an acute respiratory infection in horses worldwide, and virus neutralization (VN) has been the standard method for the detection of ERAV antibody in horse serum. Previous studies have identified recombinant virion protein VP1 (rVP1) purified under native conditions to be of high potential for the development of a diagnostic ERAV enzyme-linked immunosorbent assay (ELISA). This study presents an optimized protocol for the expression and purification of native full-length rVP1. Furthermore, we demonstrate that, upon denaturation, rVP1 ...
The continuous spread of West Nile virus (WNV): seroprevalence in asymptomatic horses. West Nile virus (WNV) was probably introduced in southern and northern Mexico from the USA in two independent events. Since then, WNV activity has been reported in several Mexican states bordering the USA and the Gulf of Mexico, but disease manifestations seen there in humans and equids are quite different to those observed in the USA. We have analysed WNV seroprevalence in asymptomatic, unvaccinated equids from two Mexican states where no data had been previously recorded. WNV IgG antibodies were detected in 31.6% (91/288) of equine sera from Chiapas and Puebla states (53.3% and 8.0%, respect...
Enzyme-linked immunosorbent assay using glycoprotein and monoclonal antibody for detecting antibodies to vesicular stomatitis virus serotype New Jersey. In this study, an enzyme-linked immunosorbent assay (ELISA) using glycoprotein and a monoclonal antibody (MAb) was developed for the detection of antibodies to vesicular stomatitis virus (VSV) serotype New Jersey (NJ). The glycoprotein to be used as a diagnostic antigen was extracted from partially purified VSV-NJ, and a neutralizing MAb specific to VSV-NJ was incorporated to compete with antibodies in a blocking ELISA using glycoprotein (GP ELISA). The cutoff of the GP ELISA was set at 40% inhibition, which corresponded to a virus neutralization test (VNT) titer of 32. With this threshold, th...
Immune response against equine gammaherpesvirus in Icelandic horses. Horses are hosts to two types of gammaherpesviruses, equine herpes virus (EHV) 2 and 5. While EHV-2 is ubiquitous in adult horses, EHV-5 has been less frequently described. Due to strong serological cross-reactivity, EHV-2 and -5 cannot be discriminated in broad spectrum antibody tests and are thus commonly referred to as gamma-EHV. Total IgG and IgG subclass response against gamma-EHV were determined in serum from 41 healthy Icelandic horses, thereof 20 adults, 10 foals aged 10 months, and 11 foals aged 1-4 months. Additionally, in 10 of the adult horses, interferon (IFN)-gamma and interleuki...
Antibody immobilization on to polystyrene substrate–on-chip immunoassay for horse IgG based on fluorescence. A simple microfluidic immunoassay card was developed based on polystyrene (PS) substrate for the detection of horse IgG, an inexpensive model analyte using fluorescence microscope. The primary antibody was captured onto the PS based on covalent bonding via a self-assembled monolayer (SAM) of thiol to pattern the surface chemistry on a gold-coated PS. The immunosensor chip layers were fabricated from sheets by CO(2) laser ablation. The functionalized PS surfaces after each step were characterized by contact angle measurement, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (...
Development of an indirect ELISA for the diagnosis of equine piroplasmosis. An indirect ELISA (iELISA) for the detection of specific anti-Theileria equi antibodies in horse serum was developed. Its performance showed good concordance (K= 0.79) when compared with a competitive ELISA recommended by the World Organisation for Animal Health. Horse serum samples from two provinces located in the north and east of Argentina (Formosa and Entre Rios, respectively) were analyzed by this iELISA. A high percentage of positive horses were found in Formosa, consistent with the climatic conditions of the region that are apt for the development of tick vectors. Surprisingly, seropos...
Serologic response to West Nile virus vaccination in the greater one-horned rhinoceros (Rhinoceros unicornis). Vaccination has been an important component of preventative health care programs of North American zoologic institutions in their protection of valuable species against West Nile virus (WNV) infection since its detection in 1999. Although approved only for horses, commercial WNV vaccine has been used for the purpose of protection of nondomestic species, including avian, equid, and rhinoceros species. Currently, there are two commercial equine vaccines available, a killed vaccine and a recombinant viral-vectored vaccine. Both products have been used for the vaccination of Greater One-horned rhi...
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...
Virion associated proteins of equine rhinitis B virus 1 (ERBV1): the non-structural protein 3C(pro) co-purifies with virions. Equine rhinitis B virus (ERBV), genus Erbovirus, is most closely related to the Cardiovirus genus in the family Picornaviridae. The structural proteins (VP1-4) of erboviruses are not well described, but are predicted by sequence to be 35, 29, 26 and 7 kDa. Methods for the purification of cardioviruses (polyethylene glycol, trypsin treatment) were used to characterise the structural proteins of ERBV1. Only one of the virus proteins detected was an expected molecular mass, and this 26 kDa protein was identified as VP3 by N-terminal amino acid sequencing. N-terminal sequencing of the 56 and a 29 ...
Antigenic, microbicidal and antiparasitic properties of an l-amino acid oxidase isolated from Bothrops jararaca snake venom. Venoms from the bee Apis mellifera, the caterpillar Lonomia achelous, the spiders Lycosa sp. and Phoneutria nigriventer, the scorpions Tityus bahiensis and Tityus serrulatus, and the snakes Bothrops alternatus, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi, Crotalus durissus terrificus, and Lachesis muta were assayed (800mug/mL) for activity against Staphylococcus aureus. Venoms from B. jararaca and B. jararacussu showed the highest S. aureus growth inhibition and also against other Gram-positive and Gram-negative bacteria. To characterize the microbicidal compon...
Serosurveillance for Japanese encephalitis, Akabane, and Aino viruses for Thoroughbred horses in Korea. Recent global warming trends may have a significant impact on vector-borne viral diseases, possibly affecting vector population dynamics and disease transmission. This study measured levels of hemagglutination-inhibition (HI) antibodies against Japanese encephalitis virus (JEV) and neutralizing antibodies against Akabane virus (AKAV) and Aino virus (AINV) for Thoroughbred horses in Korea. Blood samples were collected from 989 racehorses in several provinces, between October 2005 and March 2007. Sera were tested using either an HI assay or a virus neutralization test. Approximately half (49.7%;...
Suppression of testicular function and sexual behavior by vaccination against GnRH (Equity) in the adult stallion. The aim of this study was to evaluate the effects of an anti-GnRH vaccine on testosterone concentration, antibody titer, scrotal width, semen quality and sexual behavior in the stallion. Adverse reactions to the vaccine were also determined. Eight clinically healthy sexually experienced stallions aged between 6 and 15 years from the National Stud in Avenches (Switzerland) were used. Five stallions were immunized 3 times at an interval of 4 and 8 weeks, respectively, with 200 microg of a GnRH-protein conjugate (Equity, CSL Limited, Australia) intramuscularly in the neck and 3 control animals re...
A glycosylated peptide in the West Nile virus envelope protein is immunogenic during equine infection. Using a monoclonal antibody directed to domain I of the West Nile virus (WNV) envelope (E) protein, we identified a continuous (linear) epitope that was immunogenic during WNV infection of horses. Using synthetic peptides, this epitope was mapped to a 19 aa sequence (WN19: E147-165) encompassing the WNV NY99 E protein glycosylation site at position 154. The inability of WNV-positive horse and mouse sera to bind the synthetic peptides indicated that glycosylation was required for recognition of peptide WN19 by WNV-specific antibodies in sera. N-linked glycosylation of WN19 was achieved through ...
Horses experimentally infected with Sarcocystis neurona develop altered immune responses in vitro. Equine protozoal myeloencephalitis (EPM) due to Sarcocystis neurona infection is 1 of the most common neurologic diseases in horses in the United States. The mechanisms by which most horses resist disease, as well as the possible mechanisms by which the immune system may be suppressed in horses that develop EPM, are not known. Therefore, the objectives of this study were to determine whether horses experimentally infected with S. neurona developed suppressed immune responses. Thirteen horses that were negative for S. neurona antibodies in serum and cerebrospinal fluid (CSF) were randomly assig...
Effect of HCG in the presence of HCG antibodies on the follicle, hormone concentrations, and oocyte in mares. Follicle blood flow, follicular-fluid and plasma hormone concentrations, and oocyte quality were studied 30 h after an ovulation-inducing hCG treatment when the pre-ovulatory follicle was 32 mm. Mares were grouped as positive (n = 16) and negative (n = 44) for hCG antibodies before the experimental hCG treatment. Percentage of the follicle wall with blood flow signals was less (p < 0.05) in the antibody positive group than in the negative group. The concentrations of follicular-fluid oestradiol and free IGF1, and plasma oestradiol were greater (p < 0.001), and follicular-fluid progesterone (p ...
