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Topic:Antigen
Antigens are substances that can induce an immune response in horses, typically by being recognized as foreign by the immune system. These substances can include proteins, polysaccharides, or lipids, and are often components of pathogens such as bacteria, viruses, or parasites. In horses, antigens are essential for the activation of both the innate and adaptive immune responses, leading to the production of antibodies and the activation of immune cells. The study of antigens in equines encompasses understanding their structure, the mechanisms by which they are recognized by the immune system, and their role in vaccine development. This page compiles peer-reviewed research studies and scholarly articles that explore the identification, characterization, and immunological impact of antigens in equine health and disease.
Comparative Evaluation of a Multistrain Indirect ELISA Targeting Anti- p26 and gp45 Antibodies for EIAV Detection. Equine Infectious Anemia Virus (EIAV), a lentivirus marked by considerable genetic variability, poses significant diagnostic challenges. Existing diagnostic tools encompass the Agar Gel Immunodiffusion Assay (AGID), enzyme-linked immunosorbent assay (ELISA), and Western blotting (WB). ELISA and AGID mainly utilize the p26 capsid protein, often sourced from the Wyoming reference strain. To broaden the range of viral proteins and strains employed in these immunoassays, we previously developed a novel p26/double-strain gp45 indirect ELISA. In this study, we evaluated the performance of this ELISA...
Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4). Equine rhinopneumonia, caused by equine herpesvirus types 1 and 4 (EHV-1 and EHV-4), continues to be a significant health and economic concern in the global equine industry, particularly in Kazakhstan. While vaccines targeting EHV-1 are available, there is currently no licensed monovalent vaccine for EHV-4, and existing formulations offer limited protection against this serotype. This study aimed to develop and evaluate a freeze-dried, live-attenuated EHV-4 vaccine with improved safety, stability, and immunogenicity. Methods: A field isolate of EHV-4 was attenuated through serial passaging in ...
Analysis of distinct variants of immunoglobulin G binding protein EAG on humoral immunity and bacterial clearance of Streptococcus equi subspecies equi. Streptococcus equi subspecies equi (S. equi) causes strangles, one of the most prevalent and highly contagious equine infectious diseases with significant welfare and economic impacts. Alpha 2-macroglobulin and immunoglobulin G binding protein (EAG) has been identified as a key antigen and plays a crucial role in the immune evasion of S. equi. In this study, we aimed to investigate the genetic polymorphism of EAG and to determine the effects of genetic variation on its function and immunogenicity. Methods: Phylogenetic analysis indicated these EAG mutants belong to two clades, respectively. Th...
Serodiagnosis of equine infectious anemia by indirect ELISA based on a novel synthetic peptide derived from gp45 glycoprotein. The transmembrane glycoprotein gp45 (implicated in virus-equine B lymphocyte binding and in subsequent infection) is an important protein used as an antigen in the serological diagnosis of equine infectious anemia, but synthetic peptides derived from its structure are not. Consequently, the present study aimed to determine the diagnostic performance of the Bio-AIELAB immunoassay (novel synthetic peptide P05/gp45) in comparison with a similar commercial assay (ELISA EIA, VMRD Inc, USA) and with the gold standard (AGID, LABIOFAM, Cuba). A total of 365 samples of sera from naturally infected Cuba...
Immunoaffinity chromatography for the preparation of equine tetanus immunoglobulin F(ab’)2 for enhanced safety and efficacy. Typically, the antigen-specific antibodies constitute a small fraction-often estimated to be around 2-10 %-of the total IgG in the serum after immunization. This low percentage necessitates the use of purification techniques to enrich the antigen-specific antibodies for therapeutic or research purposes. This study introduces an affinity chromatography column using NHS-activated Sepharose as a matrix and the tetanus toxin subunit C, TeNT-Hc-C869A, as a ligand, enabling the purification of polyclonal antibodies with high specificity. This process improves antitoxin purity to over 95 %, effecti...
Seroprevalence of IgG Antibodies Against Borrelia burgdorferi Sensu Lato, Anaplasma phagocytophilum, and Tick-Borne Encephalitis (TBE) Virus in Horses in Southern Norway. ticks play a crucial role as carriers of diseases, transmitting pathogens to vertebrate hosts, including horses. This study aimed to investigate the seroprevalence of IgG antibodies against sensu lato (s. l.), , and tick-borne encephalitis virus (TBE-virus) in equine sera collected in southern Norway. In total, sera from 331 horses stabled in four counties (Agder, Vestfold and Telemark, Vestland, and Viken) were analyzed by immunoblot. In total, 66% of the horses were IgG-seropositive for antibodies against one or multiple tick-borne pathogens. The highest seroprevalence was detected against...
A comparative evaluation of seven commercial human influenza virus antigen detection kits for the diagnosis of equine influenza. Equine influenza virus (EIV) is one of the most important pathogens causing respiratory signs in equids. Rapid antigen detection (RAD) kits are useful for point-of-care testing because they are user-friendly and provide fast results. Although sensitive and broad-reactive RAD kits are needed for controlling horse movement, no RAD kits specified for EIV are available. Objective: This study evaluated the usefulness of seven RAD kits originally developed for human influenza and available in Japan during 2023-2024 for EIV antigen detection. Methods: Experimental assay comparison. Methods: The detec...
Exploration of the Differential Expression Patterns of Immunoglobulin Heavy Chain Genes in Horses and Donkeys. This study investigated the immunoglobulins (IG) gene locus structure and expression diversity in local Chinese horse and donkey breeds, including Ningqiang, Guanzhong horses, and varieties such as Guanzhong, Jiami, and Northern Shaanxi donkey using genome alignment and high-throughput sequencing. The aim was to expand understanding of IG expression patterns in horses, donkeys, and their different breeds. The results revealed that the donkey IGH locus contains 117 VH genes (23 functional), 44 DH genes, and 8 JH genes, arranged in a VH-DH-JH sequence on chromosome 7, spanning approximately 1189...
Equine tapeworm infections in Italy: A nationwide coprological survey. Grazing horses are exposed to various intestinal helminths, with tapeworms being of major concern due to their potential pathogenicity. Few regional data are available in Italy on tapeworm infections in horses; therefore, the aim of this study was to investigate the prevalence of Anoplocephala spp. eggs over a nine-year period (2014-2023) in 12,056 faecal samples from horses living in all 18 continental Italian regions. Coprological examinations were performed using the Proudman and Edwards (1992) method using Sheather's sugar solution as the floatation medium. An overall prevalence of 3.7 % ...
Comparison of immunogenicity of 17 Burkholderia mallei antigens and whole cell lysate using indirect ELISA. Glanders is a World Organization for Animal Health (WOAH)-notifiable equine disease caused by the infection of Burkholderia mallei, and is endemic in Mongolia, South Asia, Africa, and South America. While the complement fixation test (CFT) has been widely used for serodiagnosis of glanders and is considered a standard serological test, it has several limitations. These limitations include poor specificity, labor intensive techniques, variability in antigen and protocol. Consequently, indirect enzyme-linked immunosorbent assays (iELISAs) based on recombinant proteins have been developed as alte...
Detection of equid alphaherpesvirus 1 in serum samples collected from infected horses. Equid alphaherpesvirus 1 (EqAHV1; Orthoherpesviridae, Varicellovirus equidalpha1) spreads by viremia to susceptible organs. Because EqAHV1 circulates in the bloodstream in a cell-associated manner, serum samples are not considered valuable for detecting EqAHV1 and have therefore not been tested by highly sensitive detection methods such as real-time PCR (rtPCR). We investigated whether EqAHV1 could be detected by this method in equine serum samples. We performed rtPCR on archived sera and peripheral blood mononuclear cells (PBMCs) collected from 3 horses experimentally inoculated with EqAHV1. ...
Development and Validation of a Differentiating Infected from Vaccinated Animals (DIVA) Enzyme-Linked Immunosorbent Assay (ELISA) Strategy for Distinguishing Between Hendra-Infected and Vaccinated Horses. Hendra virus (HeV) is a bat-borne zoonotic agent which can cause a severe and highly fatal disease and can be transferred from animals to humans. It has caused over 100 deaths in horses since it was discovered in 1994. Four out of seven infected humans have died. Since the release of the HeV vaccine (Equivac® HeV Hendra Virus Vaccine for Horses, Zoetis Australia Pty Ltd., Rhodes, NSW 2138) in Australia, there has been an urgent requirement for a serological test for differentiating infected from vaccinated animals (DIVA). All first-line diagnostic serological assays at the Australian Centre f...
Equine Influenza: Epidemiology, Pathogenesis, and Strategies for Prevention and Control. Equine influenza (EI) is a highly contagious respiratory disease caused by the equine influenza virus (EIV), posing a significant threat to equine populations worldwide. EIV exhibits considerable antigenic variability due to its segmented genome, complicating long-term disease control efforts. Although infections are rarely fatal, EIV's high transmissibility results in widespread outbreaks, leading to substantial morbidity and considerable economic impacts on veterinary care, quarantine, and equestrian activities. The H3N8 subtype has undergone significant antigenic evolution, resulting in the...
Differential Expression of ARG1 and MRC2 in Retinal Müller Glial Cells During Autoimmune Uveitis. Retinal Müller glial cells (RMG) play a crucial role in retinal neuroinflammation, including autoimmune uveitis. Increasing evidence supports their function as active modulators of immune responses and potential atypical antigen-presenting cells (APCs). To further investigate this hypothesis, we conducted a differential proteome analysis of primary equine RMG from healthy controls and horses with equine recurrent uveitis (ERU), a spontaneous model of autoimmune uveitis. This analysis identified 310 proteins with differential abundance. Among these, the Major Histocompatibility Complex (MHC) c...
Innovative production of highly potent equine neutralizing antibody against Hemiscorpius lepturus scorpion venom using recombinant mPLD1 protein. Scorpion envenomation, especially from Hemiscorpius lepturus, poses a significant health risk, leading to considerable morbidity and mortality. The venom's major toxin, which includes phospholipase D (PLD), is responsible for various systemic complications. In prior studies, we identified a native phospholipase D (PLD) toxin as a key lethal factor in the venom of H. lepturus. A recombinant PLD that retained its toxicity was developed and designated as PLD1. Additionally, a non-toxic and devoid of lethal effects mutant form of the recombinant PLD1 protein, was produced and named as mPLD1. Build...
Diagnostic innovations in Equine Parasitology: a Nanogold-ELISA for sensitive serodiagnosis of migratory strongylus vulgaris larvae infections. Strongylus vulgaris, a devastating parasitic nematode in equids, causes life-threatening verminous aneurysms that are challenging to diagnose early. This study pioneered integrating nanotechnology into an indirect enzyme-linked immunosorbent assay (i-ELISA) system to enhance the sensitivity and specificity for detecting S. vulgaris larval antigens in equine serum samples, with PCR confirmation of the species. A conventional i-ELISA and an innovative nano-based ELISA were developed using excretory-secretory antigens from adult S. vulgaris worms. The nano-ELISA incorporated gold nanoparticles (1...
A comparative study evaluating three line immunoassays available for serodiagnosis of equine Lyme borreliosis: Detection of Borrelia burgdorferi sensu lato-specific antibodies in serum samples of vaccinated and non-vaccinated horses. Diagnosis of equine Lyme borreliosis (LB), an infection caused by members of the Borrelia burgdorferi sensu lato complex (Bbsl), is challenging due to the nonspecific clinical signs of the disease and due to the variety of non-standardized serological tests. Specific vaccine-induced antibodies against LB, providing an effective protection against the infection, complicate the issue further. The standard for the detection of specific antibodies against Bbsl is a two-tier test system based on an enzyme-linked immunosorbent assay (ELISA) or indirect fluorescent antibody test (IFA) for antibody sc...
Functional assessment of the glycoproteins of a novel Hendra virus variant reveals contrasting fusogenic capacities of the receptor-binding and fusion glycoproteins. A novel Hendra virus (HeV) genotype (HeV genotype 2 [HeV-g2]) was recently isolated from a deceased horse, revealing high-sequence conservation and antigenic similarities with the prototypic strain, HeV-g1. As the receptor-binding (G) and fusion (F) glycoproteins of HeV are essential for mediating viral entry, functional characterization of emerging HeV genotypic variants is key to understanding viral entry mechanisms and broader virus-host co-evolution. We first confirmed that HeV-g2 and HeV-g1 glycoproteins share a close phylogenetic relationship, underscoring HeV-g2's relevance to global he...
Use of equine H3N8 hemagglutinin as a broadly protective influenza vaccine immunogen. Development of an efficacious universal influenza vaccines remains a long-sought goal. Current vaccines have shortfalls such as mid/low efficacy and needing yearly strain revisions to account for viral drift/shift. Horses undergo bi-annual vaccines for the H3N8 equine influenza virus, and surveillance of sera from vaccinees demonstrated very broad reactivity and neutralization to many influenza strains. Subsequently, vaccinating mice using the equine A/Kentucky/1/1991 strain or recombinant hemagglutinin (HA) induced similar broadly reactive and neutralizing antibodies to seasonal and high path...
Immunization but not natural infection of horses results in antibody activity against the S protein of Streptococcus equi subsp equi. Evaluate the immunogenicity of a vaccine targeting the S protein (Ssee) of Streptococcus equi subsp equi and determine antibody activity against Ssee in horses with strangles. Methods: The study was designed as a prospective experiment using 20 university-owned Quarter Horses and a cross-sectional serosurvey of 78 privately owned horses with strangles. Horses were immunized IM with 0 (n = 4), 200 (n = 8), or 400 (n = 8) μg of recombinant Ssee at weeks 0, 4, and 12. Serum and nasal secretions were collected at weeks 0, 4, 6, 12, 16, and 28 and tested by ELISA for immunoglobulin (Ig)-G against ...
Immunization but not natural infection of horses results in antibody activity against the S protein of Streptococcus equi subsp equi. Evaluate the immunogenicity of a vaccine targeting the S protein (Ssee) of Streptococcus equi subsp equi and determine antibody activity against Ssee in horses with strangles. Methods: The study was designed as a prospective experiment using 20 university-owned Quarter Horses and a cross-sectional serosurvey of 78 privately owned horses with strangles. Horses were immunized IM with 0 (n = 4), 200 (n = 8), or 400 (n = 8) μg of recombinant Ssee at weeks 0, 4, and 12. Serum and nasal secretions were collected at weeks 0, 4, 6, 12, 16, and 28 and tested by ELISA for immunoglobulin (Ig)-G against ...
Evidence of Influenza A(H5N1) Spillover Infections in Horses, Mongolia. Recent outbreaks of influenza A(H5N1) have affected many mammal species. We report serologic evidence of H5N1 virus infection in horses in Mongolia. Because H3N8 equine influenza virus is endemic in many countries, horses should be monitored to prevent reassortment between equine and avian influenza viruses with unknown consequences.
Attenuated Getah virus confers protection against multiple arthritogenic alphaviruses. Alphaviruses are important arthropod-transmitted pathogens of humans and livestock. Getah virus (GETV) is an arthritogenic alphavirus that causes disease in horses and piglets; it also poses a potential threat to humans. A live attenuated vaccine candidate named GETV-3ΔS2-CM1, harbouring a deletion in nonstructural protein 3 and substitutions in the capsid protein, is genetically stable and exhibits robust immunogenicity. It was shown to confer passive protection to piglets born to immunized sows. In mice, a single dose of GETV-3ΔS2-CM1 protected against infection with different strains of G...
Antibody isotyping and cytokine profiling in natural cases of Burkholderia mallei infection (glanders) in equines. Immunological aspects of B. mallei infection were rarely studied in natural cases of equines. The present study was conducted to determine IgG, IgM, IgA and IgG (T) titre against recombinant Hcp1, TssA and TssB proteins and PilA-Hcp1-TssN-BipD and BpaB-BpaC- BMAA0553 chimeras and cytokine responses in glanders affected equine serum. Methods: The study was conducted on serum samples collected from 151 glanders positive equines which include horses (n = 134), mules (n = 16), and donkeys (n = 01). The IgM, IgG, IgA and IgG (T) titre were determined against recombinant antigens by indirect E...
A polygeneric immunogen composed of 22 venoms from sub-Saharan African snakes to expand the neutralization scope of the EchiTAb-plus-ICP antivenom. Recent research suggests that a polygeneric immunogen made from the venoms of the most medically important viperid and elapid snakes in sub-Saharan Africa could elicit a broader antibody response in horses compared to the current EchiTAb-plus-ICP antivenom, especially against neurotoxic elapid venoms. To test this, 25 horses that have been regularly immunized to produce this antivenom were reimmunized with an immunogen containing 22 venoms from various snake species from the genera , , , and both spitting and non-spitting . The plasma collected from these horses was processed using the capryli...
Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against African Horse Sickness Virus Serotype 1 VP2 Protein. African horse sickness (AHS) is an acute, fatal, contagious disease of animals of the family Equidae and is caused by infection with the African horse sickness virus (AHSV). Based on the outer capsid protein VP2, AHSV is classified into nine serotypes (AHSV-1 to -9) with little or no serological cross-reactivity between them. In 2020, AHS outbreaks caused by AHSV-1 were reported in Thailand and Malaysia, marking the first occurrences of AHS in Southeast Asia. However, little is known about the antigenic profile of AHSV-1 VP2. In this study, a recombinant VP2 protein was expressed in and used ...
Protein profile of extracellular vesicles derived from adult Parascaris spp. Parascaris spp. represent a significant threat to equine health worldwide, particularly in foals. The long-term survival of parasites in the host necessitates persistent modulation of the host immune response. Intercellular communication achieved through the exchange of molecules via extracellular vesicles (EVs) released from the parasite could be a crucial factor in this regard. This study aimed to isolate and characterize EVs released by adult male and female Parascaris worms and conduct a proteomic analysis to identify sex-specific proteins and potential immunomodulatory factors. Methods: L...
Allergen immunotherapy using recombinant Culicoides allergens improves clinical signs of equine insect bite hypersensitivity. Insect bite hypersensitivity (IBH) is an IgE-mediated allergic dermatitis of horses caused by bites of spp., sharing some common features with human atopic dermatitis. Allergen immunotherapy (AIT) using whole-body extracts has limited efficacy. This study aimed to evaluate AIT with a pool of major recombinant allergens in a prospective, double-blinded, placebo-controlled study. Unassigned: The IBH lesion score was assessed during a pre-treatment year and first treatment year (May-October) in 17 horses and in May and July of a second treatment year. Nine horses were immunized subcutaneously ...
A platform for the rapid screening of equine immunoglobins F (ab)2 derived from single equine memory B cells able to cross-neutralize to influenza virus. Single B cells-based antibody platforms offer an effective approach for the discovery of useful antibodies for therapeutic or research purposes. Here we present a method for screening equine immunoglobins F(ab)2, which offers the potential advantage of reacting with multiple epitopes on the virus. Using equine influenza virus (EIV) as model, a hemagglutinin (HA) trimer was constructed to bait B cells in vaccinated horses. We screened 370 HA-specific B cells from 1 × 10 PBMCs and identified a diverse set of equine variable region gene sequences of heavy and light chains and then recombined wit...
Identification of three novel linear B-cell epitopes on VP7 of African horse sickness virus using monoclonal antibodies. African horse sickness (AHS) is an acute and subacute infectious disease of equine species caused by the African horse sickness virus (AHSV). The VP7 of AHSV is a group-specific protein conserved in all serotypes and is an excellent candidate for the serological diagnosis and an AHS vaccine component. However, to date, B-cell epitopes on the AHSV VP7 recognized by humoral immune responses remain unclear. This study expressed the recombinant AHSV VP7 soluble in Escherichia coli and purified it for mouse immunization. Four monoclonal antibodies (mAbs) were screened and identified by hybridoma ce...
