The equine immune system is a complex network of cells, tissues, and organs that work collaboratively to defend against pathogens and maintain homeostasis. It consists of innate and adaptive components, each with distinct functions and mechanisms. The innate immune system provides the first line of defense through physical barriers, phagocytic cells, and the complement system. The adaptive immune system involves lymphocytes, such as B cells and T cells, which generate specific responses to antigens and provide immunological memory. Research in equine immunology explores the interactions between these components, the impact of genetic and environmental factors on immune function, and the development of vaccines and therapeutics. This page gathers peer-reviewed studies and scholarly articles focusing on the mechanisms, regulation, and clinical applications of the equine immune system in health and disease.
Cook RF, Cook SJ, Even DL, Schaffer C, Issel CJ.Horse IL-7 (HIL-7) cDNA was isolated from adult lymph node tissue by reverse transcription polymerase chain reaction (RT-PCR) using oligonucleotide primers based on horse genomic sequences (The Broad Institute). In addition, to the full-length (FL) 531bp reading frame encoding 176 amino acids, shorter open-reading frames of 477, 396 and 264bp were also amplified. Nucleotide sequence analysis of these RT-PCR products demonstrated they were homologous except the shorter species were missing internal sequences consistent with multiple RNA splicing events. Consequently, the shorter open-reading fr...
Bai Y, Tong TG, Zhang WJ, Xu SL, Wang Q, Liu GL, Wu DL.To develop a quantitative ELISA by measuring interferon (IFN-gamma) of equine lymphocytes. Methods: Sandwich ELISA for equine IFN-gamma was developed using mAb A5 as a capture antibody and biotinylated mAb SB10 as a detection antibody. Results: The detection limit of the sandwich ELISA for equine IFN-gamma was 1 microg/L and did not show cross-reactivity with recombinant equine IL-18. Equine IFN-gamma was detected by ELISA in culture medium of the peripheral blood mononuclear cells (PBMCs) stimulated with ConA or PMA/Ionomycin. Conclusions: This method can be used to help understand the role o...
Chang DC, Liu WJ, Anraku I, Clark DC, Pollitt CC, Suhrbier A, Hall RA, Khromykh AA.DNA vaccines encoding replication-defective viruses are safer than inactivated or live attenuated viruses but may fail to stimulate an immune response sufficient for effective vaccination. We augment the protective capacity of a capsid-deleted flavivirus DNA vaccine by co-expressing the capsid protein from a separate promoter. In transfected cells, the capsid-deleted RNA transcript is replicated and translated to produce secreted virus-like particles lacking the nucleocapsid. This RNA is also packaged with the help of co-expressed capsid protein to form secreted single-round infectious particl...
Deeg CA, Hauck SM, Amann B, Pompetzki D, Altmann F, Raith A, Schmalzl T, Stangassinger M, Ueffing M.Equine recurrent uveitis (ERU) is an autoimmune disease that occurs with a high prevalence (10%) in horses. ERU represents the only reliable spontaneous model for human autoimmune uveitis. We already identified and characterized novel autoantigens (malate dehydrogenase, recoverin, CRALBP) by analyzing the autoantibody-binding pattern of horses affected by spontaneous recurrent uveitis (ERU) to the retinal proteome. CRALBP also seems to be relevant to human autoimmune uveitis. Proteomic screening of vitreous and retinal samples from ERU diseased cases in comparison to healthy controls has led t...
Brandt S, Haralambus R, Shafti-Keramat S, Steinborn R, Stanek C, Kirnbauer R.Bovine papillomavirus type 1 or 2 (BPV-1, BPV-2) are accepted causal factors in equine sarcoid pathogenesis. Whereas viral genomes are consistently found and expressed within lesions, intact virions have never been detected, thus permissiveness of sarcoids for BPV-1 replication remains unclear. To reassess this issue, an immunocapture PCR (IC/PCR) was established using L1-specific antibodies to capture L1-DNA complexes followed by amplification of the viral genome. Following validation of the assay, 13 sarcoid-bearing horses were evaluated by IC/PCR. Samples were derived from 21 tumours, 4 per...
Lauková A, Simonová M, Strompfová V, Styriak I, Ouwehand AC, Várady M.Faecal samples of 122 horses (from farms in Slovakia) were examined to select enterococci to study their probiotic potential for their further use as additives. Each gram of faeces contained 1.0-5.0 cfu (log 10) of enterococci. Of the 43 isolates, 25 (58.1%) were identified as Enterococcus faecium, 3 strains were (6.9%) Enterococcus mundtii and one strain was identified as E. faecalis. Fourteen isolates were not characterized further. A significant proportion of the isolates were resistant to kanamycin, vancomycin and gentamicin. Low urease activity of enterococci dominated. The values of lact...
Daly J, Daas A, Behr-Gross ME.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) ...
Hobo S, Niwa H, Anzai T.The reactivity of the proline-glutamic acid-proline-lysine (PEPK) repetition peptide antigen in 3176 serum samples was investigated to evaluate its utility as an antigen for the serological diagnosis of strangles. The reactivity of the sera of horses infected with Streptococcus equi subspecies equi was high when the peptide had several PEPK repetitions. However, as the number of PEPK repetitions increased, the reactivity of the antigen with the sera of horses infected with Streptococcus equi subspecies zooepidemicus also increased. In horses infected experimentally with S equi, the reactivity ...
Lewis MJ, Meehan M, Owen P, Woof JM.The M protein of Streptococcus equi subsp. equi known as fibrinogen-binding protein (FgBP) is a cell wall-associated protein with antiphagocytic activity that binds IgG. Recombinant versions of the seven equine IgG subclasses were used to investigate the subclass specificity of FgBP. FgBP bound predominantly to equine IgG4 and IgG7, with little or no binding to the other subclasses. Competitive binding experiments revealed that FgBP could inhibit the binding of staphylococcal protein A and streptococcal protein G to both IgG4 and IgG7, implicating the Fc interdomain region in binding to FgBP. ...
Garvican ER, Vaughan-Thomas A, Redmond C, Clegg PD.Matrix degradation by metalloproteinases is considered a key feature in the loss of articular cartilage seen in many joint diseases. Membrane-type matrix metalloproteinase-3 (MT3-MMP) expression is elevated in human cartilage in end-stage osteoarthritis. We investigated whether MT3-MMP is similarly regulated in cartilage in two naturally occurring arthropathies in vivo and whether proinflammatory cytokines regulate its expression in vitro. MT3-MMP expression was evaluated in cartilage from horses with osteoarthritis and osteochondrosis and compared with age- and site-matched normal cartilage. ...
Lemos KR, Marques LC, Deaquino LP, Alessi AC, Machado RZ.An histochemical and immunohistochemical study was carried out to evaluate the mechanisms of immune response of horses experimentally infected by Trypanosoma evansi. For this purpose the HE histochemical stain and the avidin biotin peroxidase method were used. To determine the presence and immunoreactivity of immune cells we used anti-major histocompatibility complex II antibodies. Cellular infiltration phenotype was characterized with the aid of anti-CD3 antibody for T lymphocytes and by anti-BLA 36 antibodies for B lymphocytes. Macrophages were marked with an antibody against myeloid/histyoc...
Laha R, Sasmal NK.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...
Zhang YW, Davis EG, Blecha F, Wilkerson MJ.Innate immunity relies on a series of germline-encoded pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), to detect conserved microbial components. TLR9 is typically expressed intracellularly in immune cells such as dendritic cells and recognizes unmethylated bacterial or viral cytosine-phosphate-guanine DNA (CpG-DNA). To investigate innate immune responses through TLR9 signaling pathway in horses, we cloned and characterized equine TLR9. Protein sequence analysis shows that equine TLR9 has a typically conserved cytosolic Toll/interleukin-1 receptor (TIR) domain, three l...
Marenzoni ML, Passamonti F, Cappelli K, Veronesi F, Capomaccio S, Supplizi AV, Valente C, Autorino G, Coletti M.Fifteen unweaned thoroughbred foals, born on a stud farm to vaccinated mares, were clinically monitored during their first six months of life and repeatedly tested for equine herpesvirus type 1 (EHV-1) and equine herpesvirus type 4 (EHV-4). Nasopharyngeal swabs and blood samples were collected and screened respectively by PCR and seroneutralisation to detect the presence of the virus, explore its role as a possible cause of respiratory disease, and to assess the efficiency of the pcr for the diagnosis of this disease. The foals were divided into three groups on the basis of their clinical sign...
Borisevich IV, Potryvaeva NV, Mel'nikov SA, Evseev AA, Krasnianskiĭ VP, Maksimov VA.Immunoglobulin (Ig) against Marburg fever (MF) has been obtained from the equine serum. In terms of physicochemical and immunobiological properties, the obtained preparation corresponds to the quality of heterologous commercial immunoglobulins. The application of Marburg virus (MV) Ig with a titer of no less than 1:2048 by the emergency prevention scheme 1-2 hours after intraperitoneal inoculation of guinea pigs with MV in a dose of 20-50 LD50 protected 88-100% of the animals from death. MV Ig is recommended for emergency prevention of human MF.
Yamanaka T, Tsujimura K, Kondo T, Matsumura T.In this study, we evaluated whether five rapid antigen detection kits for human influenza could be used for the diagnosis of equine influenza (EI). Limiting dilution analyses showed that Directigen Flu A+B and ESPLINE INFLUENZA A&B-N had the highest sensitivities to equine-2 influenza viruses (EIVs) among the kits investigated. From the results of virus detection in nasal swabs taken from horses infected with EIV, these two kits could produce positive results in reasonable agreement with those obtained by virus isolation or RT-PCR, suggesting that these kits could be useful for rapid diagn...
Heskett KA, Mackay RJ.To investigate the use of a specific antibody index (AI) that relates Sarcocystis neurona-specific IgG quotient (Q(SN)) to total IgG quotient (Q(IgG)) for the detection of the anti-S neurona antibody fraction of CNS origin in CSF samples obtained from horses after intragastric administration of S neurona sporocysts. Methods: 18 adult horses. Methods: 14 horses underwent intragastric inoculation (day 0) with S neurona sporocysts, and 4 horses remained unchallenged; blood and CSF samples were collected on days - 1 and 84. For purposes of another study, some challenged horses received intermitten...
Patel JR, Heldens JG.This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause.
Menzies-Gow NJ, Bailey SR, Berhane Y, Brooks AC, Elliott J.To determine the effect of endotoxin (lipopolysaccharide [LPS]) on vasoactive mediator production by cultured equine digital vein endothelial cells (EDVECs). Methods: EDVECs obtained from forelimb digital veins of 7 healthy adult horses. Methods: EDVECs were incubated with or without LPS (1 microg/mL) for 0, 2, 4, 6, 22, and 24 hours. The EDVECs were incubated for 18 hours with LPS (10 pg/mL to 1 microg/mL) with or without ibuprofen, cycloheximide, or L-nitroarginine methyl ester. Medium concentrations of prostacyclin, cyclic guanosine monophosphate, endothelin-1, and thromboxane A(2) were det...
Rüegg SR, Heinzmann D, Barbour AD, Torgerson PR.For the evaluation of the epidemiology of Theileria equi and Babesia caballi in a herd of 510 horses in SW Mongolia, several mathematical models of the transmission dynamics were constructed. Because the field data contain information on the presence of the parasite (determined by PCR) and the presence of antibodies (determined by IFAT), the models cater for maternal protection with antibodies, susceptible animals, infected animals and animals which have eliminated the parasite and also allow for age-dependent infection in susceptible animals. Maximum likelihood estimation procedures were used...
Posthuma CC, Pedersen KW, Lu Z, Joosten RG, Roos N, Zevenhoven-Dobbe JC, Snijder EJ.The replication/transcription complex of the arterivirus equine arteritis virus (EAV) is associated with paired membranes and/or double-membrane vesicles (DMVs) that are thought to originate from the endoplasmic reticulum. Previously, coexpression of two putative transmembrane nonstructural proteins (nsp2 and nsp3) was found to suffice to induce these remarkable membrane structures, which are typical of arterivirus infection. Here, site-directed mutagenesis was used to investigate the role of nsp3 in more detail. Liberation of the hydrophobic N terminus of nsp3, which is normally achieved by c...
Riihimäki M, Raine A, Pourazar J, Sandström T, Art T, Lekeux P, Couëtil L, Pringle J.The aim of this study was to evaluate the contribution of bronchial epithelium to airway inflammation, with focus on mRNA and protein expression of cytokines of innate immunity IL-6, IL-10 and TNF-alpha, in horses with Recurrent Airway Obstruction (RAO) during exacerbation and in remission. Results: Despite marked clinical and physiologic alterations between exacerbation and after remission in the RAO horses no differences were detected in either cytokine mRNA or protein levels. Moreover, the expression of investigated cytokines in RAO horses on pasture did not differ from controls. In compari...
Desantis S, Albrizio M, Ventriglia G, Deflorio M, Guaricci AC, Minoia R, De Metrio G.The presence of the mu-opioid receptor and the type of glycosylation in the third extra-cellular loop of this receptor was investigated in the isthmus of mare oviduct during oestrus by means of immunoblotting and immunohistochemistry combined with enzymatic (N-glycosidase F and O-glycosidase) and chemical (beta-elimination) treatments. Immunoblotting analysis showed that the mu-opioid receptor consists of two peptides with molecular weights of around 65 and 50 kDa. After N-deglycosylation with N-glycosidase F an additional immunoreactive peptide was observed at around 30 KDa. The cleavage of O...
Redwan el-RM, Fahmy A, El Hanafy A, Abd El-Baky N, Sallam SM.In view of the disadvantages of human and equine rabies immunoglobulin still there is urgent needs for safe and cost-control anti-rabies immunoglobulins especially for person who have been severely exposed (categories III) to the virus. Our attempt to produce a less immunogenic and cheaper anti-rabies immunoglobulin affordable for those people living in developing countries, has been harnessed the ovine as a bioreactor instead the horse. The animals have been intramuscular immunized, and the plasma processed with 5% caprylic acid to yield IgG with purity of 95%. Moreover, antibody apparently i...
Venner M, Markus RG, Strutzberg-Minder K, Nogai K, Beyerbach M, Klug E.In 360 samples of colostrum and 36 samples of blood of warmblood mares, the concentration of immunoglobulin G (IgG) was evaluated in the post partal period with an ELISA and the results were compared to values obtained with 2 field methods--refractometry and colostrometry. A significant correlation (p < 0.0001) was determined between ELISA and colostrometry (r = +0.88) and between ELISA and refractometry (r = +0.93). So both field-methods seem suitable for evaluation of the colostral IgG-concentration in mares. Further the kinetic of the IgG concentration in colostrum, the volume of colostr...
El Garch H, Minke JM, Rehder J, Richard S, Edlund Toulemonde C, Dinic S, Andreoni C, Audonnet JC, Nordgren R, Juillard V.Successful vaccination against West Nile virus (WNV) requires induction of both neutralizing antibodies and cell-mediated immune responses. In this study, we have assessed the ability of a recombinant ALVAC-WNV vaccine (RECOMBITEK WNV) to elicit neutralizing antibodies and virus-specific cell-mediated immune responses in horses. In addition, we examined whether prior exposure to ALVAC-WNV vaccine would inhibit B and cell-mediated immune responses against the transgene product upon subsequent booster immunizations with the same vaccine. The results demonstrated that the recombinant ALVAC-WNV va...
Kuhn C, Lucius R, Matthes HF, Meusel G, Reich B, Kalinna BH.Sarcoptic mange (or scabies) is an important skin disease which can affect a variety of species including humans, cattle, goats, sheep, horses, pigs, rabbits, and dogs. Approximately 300 million people are affected worldwide and in lifestock animals the infestation may lead to substantial economic losses caused by depression in growth and feed conversion rates. Diagnosis of Sarcoptes infestation is difficult and only a few serological tests have been developed using whole mite antigen for diagnosis of mange in animals. Here we describe the isolation and characterisation of cDNAs of several imm...
van Galen G, Cassart D, Sandersen C, Delguste C, Nollet H, Amory H, Ducatelle R.Polyneuritis equi (PNE) is a rare neurological disease in the horse. Because of the suspicion in PNE of a T-lymphocyte mediated immune response against the myelin, the objective of this study was to determine the composition of the inflammatory infiltrate in the involved nerves of 3 horses with PNE, studied retrospectively. T-lymphocytes were demonstrated in the lesions, which suggests a T-lymphocyte mediated immune response against myelin. In addition, the presence of B-lymphocytes indicated a local production of antibodies. More research, involving a prospective study, is needed to determine...
Nemoto M, Ryan E, Lyons P, Cullinane A.The molecular epidemiology of equine group A rotaviruses (RVAs) in Ireland from 2011 to 2015 was investigated. Of 438 diagnostic specimens submitted from foals with enteric disease, 102 (23.3%) were positive for RVA using an immunochromatographic assay. G genotypes were determined for 76 equine RVAs, of which 68 (89.5%) were G3 and eight (10.5%) were G14. Of 18 RVAs (12 G3 and six G14) characterised by P genotyping, all were P[12]. G3P[12] and G14P[12] were the most prevalent genotypes of RVA in foals in Ireland, similar to other countries and consistent with previous studies in Ireland from 1...
Cho HJ, Entz SC, Deregt D, Jordan LT, Timoney PJ, McCollum WH.A potent ELISA antigen was prepared from equine arteritis virus (EAV) by differential centrifugation of EAV-infected cell culture fluid, followed by solubilization of the preparation by Triton X-100 treatment. Using this antigen and a mouse monoclonal antibody against the G(L) protein of EAV, a reliable blocking ELISA (bELISA) was developed for the detection of EAV antibodies in equine sera. The bELISA was evaluated using a total of 837 test serum samples. The relative sensitivity (n = 320) of the bELISA compared to the serum neutralization (SN) test was 99.4%. The bELISA appears to be a highl...
Parma AE, Cerone SI, Sansinanea SA.The antigenic relationship between Leptospira interrogans, equine cornea and lens was previously noted in our studies. Serum antibodies from horses inoculated with serovars wolffi, pomona, icterohaemorrhagiae, and tarassovi, were able to bind to five antigenic fractions from both cornea and lens, as demonstrated by immunoblotting. These antigens seem to be made up of protein and carbohydrates. After treatment with periodate for cleavage of glycoside ring structures, those fractions kept their condition of target for anti-Leptospira antibodies. Nevertheless, all fractions lost that condition af...
McGuire TC.Comparison of immunoglobulin levels of nine horses before and after infection with equine infectious anaemia (EIA) virus demonstrated a significant depression of serum IgG(T) at 2 months (P less than 0-001) and at 1 year (P less than 0-01) after infection. In contrast, the levels of IgGa were significantly increased at both times after infection. Another sixteen horses with EIA for 1-4 months were examined and there was also significant depression (P less than 0-001) of IgG(T) when compared to pre-infection levels. No significant changes in IgG(T), IgGa and IgM were noted in fourteen normal ho...
Marlo TL, Giuliano EA, Tripathi R, Sharma A, Mohan RR.To explore the impact of equine corneal fibroblast (ECF) to myofibroblast (ECM) differentiation by altering the expression of the Smad genes either individually or in combination. Specifically, we sought to examine the ECF differentiation after (a) silencing of Smad2, 3, and 4 profibrotic genes individually and (b) overexpression of antifibrotic Smad7 gene and in a combination with pro- and antifibrotic Smad genes. Methods: Equine corneal fibroblast primary cultures were generated as previously described. ECFs were transfected with individual plasmids which silenced gene expression of either S...
Banks KL, Henson JB.A defect in lymphocyte function could be responsible for persistent infection by the equine infectious anemia virus. The number of lymphocytes bearing surface immunoglobulin, as detected by immunofluorescence, and lymphocyte response to mitogens were the same in uninfected and equine infectious anemia-infected animals. A defect in T or B lymphocyte numbers or ability to respond to stimuli was not detected in this chronic virus disease.
Marois P, Pavilanis V, Boudreault A, Di Franco E.The clinical diagnosis of equine influenza was first based on the spectacular contagiousness of the disease, the general clinical resemblances to human influenza and the almost complete absence of complications usually observed in infectious viral arteritis, viral rhinopneumonitis or in other respiratory infections of the horses. The specific viral etiology of the epizootic was ascertained through the isolation of a type A influenza virus and further substantiated by evaluation of the immunological response of the sick horses, as demonstrated by complement fixation and hemagglutination-inhibit...
Saville WJ, Dubey JP, Oglesbee MJ, Sofaly CD, Marsh AE, Elitsur E, Vianna MC, Lindsay DS, Reed SM.Sarcocystis neurona and Sarcocystis fayeri infections are common in horses in the Americas. Their antemortem diagnosis is important because the former causes a neurological disorder in horses, whereas the latter is considered nonpathogenic. There is a concern that equine antibodies to S. fayeri might react with S. neurona antigens in diagnostic tests. In this study, 4 ponies without demonstrable serum antibodies to S. neurona by Western immunoblot were used. Three ponies were fed 1 x 10(5) to 1 x 10(7) sporocysts of S. fayeri obtained from dogs that were fed naturally infected horse muscles. A...
Sentsui H, Kono Y.Horse erythrocytes treated with equine infectious anemia virus hemagglutinin were phagocytized by cultivated horse leukocytes (mainly macrophage-like cells and partly polymorphonuclear cells) after incubation with fresh horse serum but not with inactivated horse serum. The phagocytosis began as soon as the erythrocytes were added to the leukocyte cultures, and the majority of the reaction proceeded within 30 minutes. Addition of antiserum showed a slightly suppressing but no enhancing effect on the phagocytosis. Phagocytosis seemed to be caused by the recognition of the third complement compon...
Halassy B, Kurtović T, Lang Balija M, Brgles M, Tunjić M, Sviben D.The hyperimmune horse plasma (HHP), prepared through active immunisation of horses with an antigen of interest, is the most common starting material for antitoxin (animal antibody-based therapeutics) production. Precise IgG quantification in plasma is a prerequisite for accurate estimation of the purification process efficiency. Although immunoglobulins from HHP have been purified for over a century, there is still no in vitro method for precise and accurate determination of IgG content in HHP. For this reason, the purification process efficiency has been assessed by antibody activity measurem...
Jeronimo C, Archambault D.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...
Liu M, Bordin A, Liu T, Russell K, Cohen N.Focusing on the first 3 weeks of life, this study examined the mRNA transcript development of different Th-type cytokines in foals in response to Rhodococcus equi infection in vitro. Results demonstrated the significant up-regulation in expression of Th1-, Th2-, and Th17-type cytokines (IFN-γ, IL-4, IL-6, IL-8, IL-12p35, IL-12p40, IL-17, IL-23p19, and TNF-α) in R. equi infection of bronchial alveolar lavage (BAL) cells of 10-day-old foals. Consequently, signature cytokines of 3 Th cell types, IFN-γ (Th1), IL-4 (Th2), and IL-17 (Th17), were used to compare temporal response patterns of circu...
Higuchi T, Hashikura S, Gojo C, Inui T, Satoh S, Yoshida M, Ishiyama T, Yamada H, Takai S.An enzyme-linked immunosorbent assay (ELISA) for detection of serum IgG antibodies against Tween 20-extracted antigen of strain ATCC 6939 was applied in Hidaka, Japan to a total of 752 sick foals showing a variety of signs of infectious disease. An optical density (OD) value of more than 0.3 was tentatively fixed to be positive on the basis of readings made of healthy horse sera in previous studies. During a 2 year study, 138 of the 752 sick foals showed an OD value of 0.3 or higher and were designated as 'suspected of R. equi infection'. Age distribution during the initial medical examination...
Bonenberger TE, Ihrke PJ, Naydan DK, Affolter VK.Immunostaining with polyclonal anti-Mycobacterium bovis (BCG) was evaluated as a single screening method for the histological identification of micro-organisms in skin biopsy specimens from various veterinary species. Confirmed archival cases infected with Mycobacteria, Nocardia, Actinobacillus, Actinomyces, Streptococcus/Staphylococcus, Dermatophilus, spirochetes, Blastomyces, Coccidioides, Cryptococcus, Histoplasma, dermatophytes, Malassezia, Sporothrix, Leishmania, Pythium, phaeohyphomycetes and Prototheca organisms were selected. A total of 70 skin biopsy specimens from the dog, cat, horse...
Kumar S, Kumar R, Gupta AK, Yadav SC, Goyal SK, Khurana SK, Singh RK.Equine piroplasmosis is a tick-transmitted protozoan disease caused by Theileria equi and/or Babesia caballi. In the present study, we expressed a 53kDa protein from the truncated EMA-2 gene of T. equi (Indian strain) and developed EMA-2ELISA using this expressed protein. This ELISA is able to detect T. equi-specific antibodies in experimentally infected animals as early as 9 days post-infection. The assay developed was validated with the OIE recommended competitive ELISA (cELISA) on 120 serum samples and significant agreement (kappa=0.93) was observed between results of both the ELISAs which ...
Bilzer T, Grabner A, Stitz L.Tissues from nine horses and one donkey suffering from natural Borna disease were investigated. Clinically, all animals demonstrated progressive reduced mentation and aggravating gait disturbances. During the clinical course anorexia and progressive loss of proprioception were observed. Cranial nerve failure was accompanied by signs of pharyngeal paralysis, sialorrhea, bruxism, and by blindness. Virologically, infectious virus was detected in the brain of all animals investigated but was not found regularly in all areas of the brain. However, in all cases, infectivity was found in the thalamus...
Dietze B, Cierpka E, Schäfer M, Schill W, Lutz MB.Equine dendritic cells (eqDC) can be generated from peripheral blood monocytes by propagation in GM-CSF and IL-4. Despite similarities with the generation of human DC, we found significant improvements for eqDC generation and functional influences on eqDC maturation. The fractionation of peripheral blood mononuclear cells (PBMC) by two subsequent gradients at densities of 1.090 and 1.077 as well as an adherence step in AIM V((R)) medium on dishes coated with extracellular matrix components (Primaria) improved the purity and yield of DC. After 3 days, eqDC cultures with GM-CSF alone developed i...
Camire AC, O'Bier NS, Patel DT, Cramer NA, Straubinger RK, Breitschwerdt EB, Funk RA, Marconi RT.Lyme disease (LD) is a tick-transmitted bacterial infection caused by Borreliella burgdorferi and other closely related species collectively referred to as the LD spirochetes. The LD spirochetes encode an uncharacterized family of proteins originally designated rotein amily welve (PF12). In B. burgdorferi strain B31, PF12 consists of four plasmid-carried genes, encoding BBK01, BBG01, BBH37, and BBJ08. Henceforth, we designate the PF12 proteins amily welve ipoprotein (Ftl) (FtlA) (BBK01), FtlB (BBG01), FtlC (BBH37), and FtlD (BBJ08). The goal of this study was to assess the potential utility o...
Fedorka CE, Loux SL, Scoggin KE, Adams AA, Troedsson MHT, Ball BA.The tolerance of pregnancy by the maternal immune system is balanced between recognition and protection. In the human this is controlled by balancing helper T cell populations (Th1, Th2) in addition to immune suppression from the regulatory arm (Tregs), but this has not been evaluated in the horse. RNA sequencing was performed on chorioallantois and endometrium of mares at 120, 180, 300 and 330 days of gestation (n = 4/stage), as well as 45-day chorioallantois (n = 4) and diestrus endometrium (n = 3). Transcripts were selected for relativity to Th1, Th2, or Treg-associated. qPCR an...
Carpenter S, Baker JM, Bacon SJ, Hopman T, Maher J, Ellis SA, Antczak DF.Sequence and functional analyses were undertaken on two cDNAs and a genomic clone encoding horse major histocompatibility complex (MHC) class I molecules. All of the clones were isolated from a single horse that is homozygous for all known horse MHC class I and class II antigens. The two cDNAs (clones 8-9 and 1-29) were isolated from a lymphocyte library and encode polymorphic MHC antigens from two loci. The genomic cosmid clone, isolated from a sperm library, contains the 8-9 gene. All three genes were expressed in mouse L-cells and were recognized by alloantisera and, for the cDNAs, by allor...
Scopel e Silva D, de Castro CC, da Silva e Silva F, Sant'anna V, Vargas GD, de Lima M, Fischer G, Brandelli A, da Motta Ade S, Hübner Sde O.P34 is an antimicrobial peptide produced by a Bacillus sp. strain isolated from the intestinal contents of a fish in the Brazilian Amazon basin with reported antibacterial activity. The aim of this work was to evaluate the peptide P34 for its in vitro antiviral properties against canine adenovirus type 2 (CAV-2), canine coronavirus (CCoV), canine distemper virus (CDV), canine parvovirus type 2 (CPV-2), equine arteritis virus (EAV), equine influenza virus (EIV), feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1). The results showed that the peptide P34 exhibited antiviral activity a...
Janda J, Plattet P, Torsteinsdottir S, Jonsdottir S, Zurbriggen A, Marti E.Allergic horses react to innocuous environmental substances by activation of Th2 cells and production of allergen-specific IgE antibodies. The mechanisms leading to Th2 differentiation are not well understood. In humans and mice, epithelial cell-derived thymic stromal lymphopoietin (TSLP) plays a central role in this process. Little is known about equine TSLP (eqTSLP) and its role in allergic diseases and our current knowledge is limited to the assessment of TSLP mRNA expression. In order to be able to study eqTSLP at the protein level, the aim of the present study was to produce recombinant e...
van Gennip RGP, van de Water SGP, Potgieter CA, van Rijn PA.The Reoviridae family consists of nonenveloped multilayered viruses with a double-stranded RNA genome consisting of 9 to 12 genome segments. The Orbivirus genus of the Reoviridae family contains African horse sickness virus (AHSV), bluetongue virus, and epizootic hemorrhagic disease virus, which cause notifiable diseases and are spread by biting Culicoides species. Here, we used reverse genetics for AHSV to study the role of outer capsid protein VP2, encoded by genome segment 2 (Seg-2). Expansion of a previously found deletion in Seg-2 indicates that structural protein VP2 of AHSV is not essen...
Guidolin FR, Tambourgi DV, Guidolin R, Marcelino JR, Okamoto CK, Magnoli FC, Queiroz GP, Dias da Silva W.Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated components, crotapotin and phospholipase A2 (PLA2). Treatment of the victims with specific antiserum is the unique effective therapeutic measure. The ability of anti-Crotalus antisera produced by the routine using crude venom to immunize horses or purified crotoxin and PLA2 as individual immunogens was compared. Antisera obtained from horses immunized...
Considering the course of the current SARS-CoV-2 pandemic, it is important to have serological tests for monitoring humoral immune response against SARS-CoV-2 infection and vaccination. Herein we describe a novel bridge enzyme-linked immunosorbent assay (b-ELISA) for SARS-CoV-2 antibodies detection in human and other species, employing recombinant Spike protein as a unique antigen, which is produced at high scale in insect larvae. Eighty two human control sera/plasmas and 169 COVID-19 patients' sera/plasmas, confirmed by rRT-PCR, were analyzed by the b-ELISA assay. In addition, a total of 27 a...
Finno CJ, Packham AE, David Wilson W, Gardner IA, Conrad PA, Pusterla N.The purpose of this study was to determine the effect of blood contamination of cerebrospinal fluid (CSF) on the results of indirect fluorescent antibody tests (IFATs) for Sarcocystis neurona and Neospora hughesi. The in vitro study used antibody-negative CSF collected from non-neurologic horses immediately after euthanasia and blood samples from 40 healthy horses that had a range of IFAT antibody titers against S. neurona and N. hughesi. Serial dilutions of whole blood were made in seronegative CSF to generate blood-contaminated CSF with red blood cell (RBC) concentrations ranging from 10 to ...
Hondalus MK, Sweeney CR, Mosser DM.A Rhodococcus equi radiobinding assay has been developed using organisms labeled with 3H-uracil. These labeled organisms resemble their unlabeled counterparts with respect to colony morphology, viability, and buoyant density. Bacteria routinely incorporate between 5 x 10(-3) and 5 x 10(-2) counts per minute per colony forming unit (cfu) which in this assay allows the detection of fewer than 0.2 cfu per macrophage. Once incorporated, greater than 90% of the label remains bacterial associated for at least 4 h postlabeling. The majority of the label is trichloroacetic acid precipitable, partition...
Bonin CP, Baccarin RY, Nostell K, Nahum LA, Fossum C, de Camargo MM.Engagement of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) is a master trigger of the deleterious effects of septic shock. Horses and humans are considered the most sensitive species to septic shock, but the mechanisms explaining these phenomena remain elusive. Analysis of tlr4 promoters revealed high similarity among LPS-sensitive species (human, chimpanzee, and horse) and low similarity with LPS-resistant species (mouse and rat). Four conserved nuclear factor kappa B (NFκB) binding sites were found in the tlr4 promoter and two in the md2 promoter sequences that are likely to be t...
Engelhardt H, King GJ.The epitheliochorial placenta represents the least intimate association between maternal and fetal tissues. The best known examples of this form of placentation are the domestic livestock species. Current information on the nature and proposed functions of uterine lymphocyte populations in ruminants (sheep and cattle), horses and pigs is presented. In ruminants unusual gamma delta T cells may play a role in mid to late gestation. During normal horse pregnancy, fetally derived endometrial cup cells invade the uterine stroma and are destroyed by maternal leukocytes midway through gestation. Natu...
Stahl C, Unger L, Mazuet C, Popoff M, Straub R, Frey J.Botulinum neurotoxins, predominantly serotypes C and D, cause equine botulism through forage poisoning. The C-terminal part of the heavy chain of botulinum neurotoxin types C and D (HcBoNT/C and D) was expressed in Escherichia coli and evaluated as a recombinant mono- and bivalent vaccine in twelve horses in comparison to a commercially available toxoid vaccine. A three-dose subcutaneous immunization of adult horses elicited robust serum antibody response in an ELISA using the immunogen as a capture antigen. Immune sera showed dose-dependent high potency in neutralizing specifically the active...
