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Topic:Immunology
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.
Equine herpesvirus 1 glycoprotein D expressed in E. coli provides partial protection against equine herpesvirus infection in mice and elicits virus-neutralizing antibodies in the horse. The envelope glycoprotein D of EHV-1 (EHV-1 gD) is essential for virus infectivity and entry of virus into cells and is a potent inducer of virus-neutralizing antibody. In this study, truncated EHV-1 gD (gDt) was expressed with a C-terminal hexahistidine tag in E. coli using a pET vector. Western blot analysis using an anti-gD monoclonal antibody demonstrated the presence of gDt bands at 37.5, 36, 29.5 and 28 kDa. The immunogenicity and protective efficacy of partially purified gDt was compared with gD expressed in insect cells by a recombinant baculovirus (Bac gD) using a BALB/c mouse model o...
Immune escape of equine herpesvirus 1 and other herpesviruses of veterinary importance. Equine herpesvirus (EHV)-1 is a pathogen of horses, well known for its ability to induce abortion and nervous system disorders. Clinical signs may occur despite the presence of a virus-specific immune response in the horse. The current review will summarize the research, on how, EHV-1-infected cells can hide from recognition by the immune system. Research findings on immune evasion of EHV-1 will be compared with those of other herpesviruses of veterinary importance.
Detection of equine herpesvirus-specific effector and memory cytotoxic immunity in the equine upper respiratory tract. Immunological protection of horses from equine herpesvirus-1 (EHV-1) infection and disease depends on the cooperation of virus-specific humoral and cellular immune responses. EHV-specific mucosal immunity may be an important component of such immune responses. This study demonstrates the induction of anti-EHV cytotoxic cellular immune responses in various mucosal and systemic lymphoid tissues associated with the upper respiratory tract (URT) of the horse. Four young horses (1-2 years of age) were inoculated intranasally with the Army 183 strain of EHV-1 and euthanized 1 week later. One untreat...
Development of specific immunoglobulin Ga (IgGa) and IgGb antibodies correlates with control of parasitemia in Babesia equi Infection. In this study, the kinetics of specific immunoglobulin G (IgG) isotypes were characterized in Babesia equi (Theileria equi)-infected horses. IgGa and IgGb developed during acute infection, whereas IgG(T) was detected only after resolution of acute parasitemia. The same IgG isotype profile induced during acute infection was obtained by equi merozoite antigen 1/saponin immunization.
Severe combined immunodeficiency in a Fell pony foal. Five days after birth of a viable Fell pony filly, yellow watery diarrhoea appeared without any signs of systemic disease. Four days later the diarrhoea ceased. On 11th day, the animal showed apathy, and a few days later, the foal was very lethargic, suffered from muscular weakness and severe watery diarrhoea that reappeared. The illness did not respond to therapy. At the age of 21 days the filly spontaneously died under symptoms of intestinal colic and pneumonia. Haematological examinations revealed lower numbers of erythrocytes as well as non-selective lymphopenia. Phagocytic activity was sl...
A monoclonal antibody to equine interleukin-4. Interleukin-4 (IL-4) is secreted by T helper type 2 cells, mast cells, basophils and eosinophils. Detection of IL-4 can contribute the evaluation of cellular immune responses during infectious diseases, immunological disorders or vaccination. We used recombinant equine IL-4 to generate a monoclonal antibody (mAb) to equine IL-4. The mAb detected recombinant IL-4 in mammalian cells transfected with different plasmids containing IL-4 cDNA. After mitogen stimulation of equine peripheral blood mononuclear cells, an intracellular protein was recognized by the new mAb in 1-2% of lymphocytes using fl...
Immunohistochemical determination of the expression of endothelin receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease. To immunohistochemically determine the expression of endothelin (ET) receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease (SPAOPD). Methods: Tissue specimens obtained from 8 healthy and 8 SPAOPD-affected horses. Methods: Horses were examined and assigned to healthy and SPAOPD groups. Horses were then euthanatized, and tissue specimens containing bronchi of approximately 4 to 8 mm in diameter were immediately collected from all lung lobes, fixed in zinc-formalin solution for 12 hours, and embedded in...
Sequence variation of the SeM gene of Streptococcus equi allows discrimination of the source of strangles outbreaks. Improved understanding of the epidemiology of Streptococcus equi transmission requires sensitive and portable subtyping methods that can rationally discriminate between strains. S. equi is highly homogeneous and cannot be distinguished by multilocus enzyme electrophoretic or multilocus sequence-typing methods that utilize housekeeping genes. However, on sequence analysis of the N-terminal region of the SeM genes of 60 S. equi isolates from 27 strangles outbreaks, we identified 21 DNA codon changes. These resulted in the nonsynonymous substitution of 18 amino acids and allowed the assignment of...
Sensitization to horse allergens without apparent exposure to horse. Report of two cases. It has been well established that occupational exposure to horses increases the risk of allergic sensitization to horse hair, on the contrary very few data have been published in individuals without direct contact with horses. We describe two cases of severe respiratory symptoms triggered by a single direct exposure in two patients with only 4-5 previous short contacts with horses during the entire their life, since they live in urban area of Naples. Cutaneous and serological diagnostic tests showed significant sensitization to horse dander other than some common inhalant allergens. Our report...
Equine herpesvirus type 1 modified live virus vaccines: quo vaditis? Infections of horses with equine herpesvirus type 1 (EHV-1) have garnered new attention over the last few years. Devastating outbreaks occurring worldwide, primarily of the neurologic form of the disease, have resulted in a reassessment of the control strategies, and particularly the prophylactic measures, that are necessary to keep the infection and spread of disease in check. Most of the available EHV-1 vaccines are based on preparations of inactivated virus, which are applied monovalently for prevention of EHV-1-caused abortion in pregnant mares or as part of multivalent vaccines to prevent...
Inter- and intramolecular epitope spreading in equine recurrent uveitis. To test the hypothesis that inter- and intramolecular spreading to S-antigen (S-Ag) and interphotoreceptor retinoid binding protein (IRBP)-derived epitopes occurs in a spontaneous model of recurrent uveitis in the horse. Methods: The immune response of eight horses with equine recurrent uveitis (ERU) was compared with that of five control horses with healthy eyes. Lymphocytes derived from peripheral blood (PBLs) were tested every 8 weeks for their reactivity against S-Ag and various S-Ag and IRBP-derived peptides for 12 to 39 months (median, 22 months). During uveitic episodes, additional bloo...
[Recombinant antibodies: a new application in scorpion envenomation?]. Serotherapy is the only specific treatment for envenomation. The antibodies are obtained after the purification of serum from hyperimmunised horses and are used after fragmentation in the form of polyclonal Fab or F(ab)'2. The anti-venom sera are heterogeneous, and their protective effect is often weak. The administration of these preparations induces risks of immediate or delayed side effects: hypersensitivity reactions, anaphylactic shock and serum sickness. This observation led us to develop new forms of antibodies produced by molecular engineering, capable of specifically neutralizing the ...
Equine CCL11 induces eosinophil cytoskeletal reorganization and activation. To assess the biological effects of purified recombinant equine CCL11 on equine eosinophil function. Methods: Following stimulation of eosinophils from normal horses, the polymerised form of actin was measured by flow cytometry using fluorescently labelled phalloidin. Migration was determined in a 96 well plate chemotaxis assay using 8 microm pore membranes, and adherence of eosinophils to serum-coated plastic was assessed using a colorimetric assay for eosinophil peroxidase. Superoxide generation was measured by the reduction of cytochrome C in a colorimetric assay. Results: Equine CCL11 indu...
Proteomic evaluation of milk from different mammalian species as a substitute for breast milk. As milk represents the main source of nutrition for infants, the question of an effective human milk substitute becomes mandatory when a formula-fed baby is allergic to cows' milk proteins. In this case, formulas containing extensively hydrolysed milk proteins should be preferred, but even such a formula may cause allergic reactions in highly sensitive patients. If there is evidence of cows' milk allergy with IgE-associated symptoms, after 6 mo of age, a soy bean formula may be recommended only when tolerance to soy protein has been established by clinical challenge. In infants with allergic r...
Immunolocalization of aquaporin-5 expression in sweat gland cells from normal and anhidrotic horses. Western blot analysis showed that sweat gland cells from freely sweating horses expressed the water channel aquaporin-5 (AQP-5). Immunohistochemistry revealed a strong AQP-5-like activity reaction at the apical membrane of the glandular secretory cells, which was absent from the surrounding myoepithelium and all other skin structures. In anhidrotic horses, AQP-5 was also found at the apical membrane of the luminal sweat gland cells, but the level of expression reduced with the length of time that the animal had displayed anhidrosis. The level of AQP-5 expression was substantially reduced in an...
Evaluation of an in vitro sulphidoleukotriene release test for diagnosis of insect bite hypersensitivity in horses. Insect bite hypersensitivity (IBH) is an IgE-mediated allergic dermatitis caused by bites of Culicoides and Simulium species, and improved means of diagnosis are required. Objective: The cellular antigen simulation test (CAST) with C. nubeculosus and S. vittatum extracts was assessed in a population of IBH-affected and healthy horses. Variations in test results over a one year period and possible cross-reactivity between different insect extracts was studied. Methods: A total of 314 mature horses were studied using the CAST. Influence of severity of clinical signs, gender and age were evaluate...
Equine Culicoides hypersensitivity: evaluation of a skin test and of humoral response. Intradermal tests were carried out on 18 horses with clinical signs of Culicoides hypersensitivity (CHS) and 23 horses without clinical signs of CHS, and sera from these horses were analysed by SDS-PAGE and Western blotting (W-B). Intradermal injections of 0.1 ml of 25 microg/microl sterile Culicoides extract, 0.1 ml of 1:10,000 histamine (positive control) and 0.1 ml of physiological saline (negative control) were made in the dermis of the middle region of the neck. Analysis of reactions indicated that a 1 cm wheal and a skinfold thickness >10% at 24 h represented a valid cut-off between h...
[Induction of EIAV-specific cellular immune response by attenuated EIAV vaccine]. To elucidate cellular immune protective mechanism of EIAV. Methods: Four horses were immunized with (DLV) by subcutaneous injection and 2 horses with 0.85% sodium chloride as the negative control. Rectal temperatures and clinical features were recorded daily. Whole blood samples were collected, from which PBMC were separated and used for CTL assay and lymphocyte proliferation assay. Results: The target cells were activated by PWM and treated with DLV or recombinant vaccinia vectors expressing DLV and LN Gag or Env separately. The percentage of EIAV-specific CTL lysis was under 5% in the negati...
A conservative domain shared by HIV gp120 and EIAV gp90: implications for HIV vaccine design. Both HIV and EIAV belong to the retroviridae family and lentivirus genus. Two variable regions (V3 and V4) of equine infectious anemia virus (EIAV) gp90 and two variable regions (V1 and V2) of HIV gp120 possibly adopt the same topology. We have studied the N-glycosylation properties and B cell linear epitope distribution profile of these two regions. Our results indicated that V3 and V4 of EIAV gp90 are very similar to V1 and V2 of HIV gp120. The differences between EIAV virulent and vaccine strains are mainly located at these two regions. Vaccine strains lose two N-glycosylation sites at thes...
Occurrence of IgE in foals: evidence for transfer of maternal IgE by the colostrum and late onset of endogenous IgE production in the horse. IgE is the key antibody involved in type I allergies. Allergen mediated crosslinking of IgE bound to high affinity Fcepsilon-receptors on mast cells and basophils stimulates cellular degranulation and release of inflammatory mediators and cytokines. In this report, we demonstrate that IgE antibodies can be transferred from the mother to offspring in horses via the colostrum. We found a clear correlation between the IgE concentration in colostrum and the total IgE concentration in foal sera on day 2 after birth (r(sp)=0.83). Maternal IgE was detected in foal sera by ELISA and on peripheral bloo...
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...
Analysis of the horse V(H) repertoire and comparison with the human IGHV germline genes, and sheep, cattle and pig V(H) sequences. We have constructed a chimeric antibody single-chain Fv (scFv) fragments phage-displayed library that combines an invariant human V(L) chain with the repertoire of V(H) domains amplified from a horse immunized against scorpion venom. To gain insight into the equine V(H) repertoire, the V(H) sequences of 46 unique clones randomly chosen from the library prior to antigenic selection were analyzed. Comparisons with previously reported equine V(H) sequences, as well as with the repertoire of human IGHV germline genes and known V(H) sequences of sheep, cattle and pig, suggest that the equine IGH lo...
Distribution of eosinophils and mast cells in the cervical tissue of non-gravid mares during dioestrus. Little is known about the local cellular immune system of the equine cervix. In this study we characterize the distribution of eosinophilic granulocytes and mast cells in the cervical tissue of non-gravid mares during dioestrus. For this purpose cervices of 10 mares were histologically examined after Sirius red and Toluidine blue staining. Both cell types could be found in the lamina propria mucosae. In the tunica muscularis the two cell populations were only rarely detected. No cells could be found in the epithelium mucosae. There was a decreasing cell density from the vaginal to uterine side...
Evaluation of five commercially available assays and measurement of serum total protein concentration via refractometry for the diagnosis of failure of passive transfer of immunity in foals. To determine and compare sensitivity, specificity, accuracy, and predictive values of measurement of serum total protein concentration by refractometry as well as 5 commercially available kits for the diagnosis of failure of passive transfer (FPT) of immunity in foals. Methods: Prospective study. Methods: 65 foals with various medical problems and 35 clinically normal foals. Methods: IgG concentration in serum was assessed by use of zinc sulfate turbidity (assay C), glutaraldehyde coagulation (assay D), 2 semiquantitative immunoassays (assays F and G), and a quantitative immunoassay (assay H)....
Replication of West Nile virus in equine peripheral blood mononuclear cells. A cell model of primary monocytes and other mononuclear cells isolated from equine blood was used to study the kinetics of West Nile virus (WNV) replication in a natural host. West Nile virus has emerged on the North American continent as a significant cause of morbidity and mortality in a wide range of avian and mammalian species. While other flaviviruses are known to infect monocytes and lymphocytes, the ability of WNV to productively replicate in specific immune cells of peripheral blood has not been assessed. In this study, enriched populations of monocytes and lymphocytes as well as purif...
Cytogenetic mapping of immunity-related genes in the domestic horse. Chromosomal locations of 19 horse immunity-related loci (CASP1, CD14, EIF5A, FCER1A, IFNG, IL12A, IL12B, IL12RB2, IL1A, IL23A, IL4, IL6, MMP7, MS4A2, MYD88, NOS2A, PTGS2, TFRC and TLR2) were determined by fluorescence in situ hybridization. For IFNG and PTGS2, this study is a confirmation of their previously reported position. In addition, microsatellite (HMBr1) was localized in the same region as IFNG. All genes were assigned to regions of conserved synteny and the data obtained in this study enhance the comparative human-horse map. Cytogenetic localization of IL6 to ECA4q14-q21.1 suggested a...
Differential susceptibility of equine and mouse brain microvascular endothelial cells to equine herpesvirus 1 infection. Equine herpesvirus 1 (EHV-1) shows endotheliotropism in the central nervous system (CNS) of infected horses. However, infection of endothelial cells has not been observed in the CNS of infected mice. To explore the basis for this difference in endotheliotropism, we compared the susceptibility of equine brain microvascular endothelial cells (EBMECs) and mouse brain microvascular endothelial cells (MBMECs) to EHV-1 infection. The kinetics of viral growth in EBMECs was typical of a fully productive infection whereas viral infection in MBMECs seemed to be nonproductive. Immunofluorescence microsco...
Endocytosis and a low-pH step are required for productive entry of equine infectious anemia virus. Recently, it has become evident that entry of some retroviruses into host cells is dependent upon a vesicle-localized, low-pH step. The entry mechanism of equine infectious anemia virus (EIAV) has yet to be examined. Here, we demonstrate that wild-type strains of EIAV require a low-pH step for productive entry. Lysosomotropic agents that inhibit the acidification of internal vesicles inhibited productive entry of EIAV. The presence of ammonium chloride (30 mM), monensin (30 microM), or bafilomycin A (50 nM) in the medium dramatically decreased the number of EIAV antigen-positive cells. We foun...
