Topic:Immunoglobulins
Immunoglobulins in horses are antibodies produced by the immune system to identify and neutralize pathogens such as bacteria and viruses. These proteins are essential components of the horse's immune response and are involved in recognizing foreign antigens. The primary classes of immunoglobulins in horses include IgG, IgA, IgM, IgE, and IgD, each with distinct roles in immune function. IgG is the most abundant and plays a key role in systemic immune responses, while IgA is important for mucosal immunity. IgM is the first antibody produced in response to an antigen, and IgE is involved in allergic reactions. IgD's function is less well-defined but is thought to be involved in respiratory immune responses. This page aggregates peer-reviewed research studies and scholarly articles that explore the structure, function, and clinical relevance of immunoglobulins in equine health.
Improved hepatic and pancreatic localisation of the equine alpha-1-proteinase inhibitor family of serpins using an antigen enhancement technique and a monoclonal antibody. Equine alpha-1-proteinase inhibitor (API) consists of three, occasionally four, serum glycoproteins. This study investigated the immunohistochemical localisation of equine API in paraformaldehyde fixed, paraffin embedded equine tissue samples of liver, lung, stomach, pancreas, jejunum and colon in five horses using affinity purified sheep polyclonal and protein A purified mouse monoclonal antibodies, whose specificities were verified by Western blotting. Exposing tissue sections to boiling citrate buffer greatly enhanced antigen recovery and improved immunostaining with both antibodies, result...
Suppressant effect of human or equine rabies immunoglobulins on the immunogenicity of post-exposure rabies vaccination under the 2-1-1 regimen: a field trial in Indonesia. MAS054 Clinical Investigator Group. WHO's reference protocol for post-exposure rabies vaccination advises five intramuscular injections on days 0, 3, 7, 14, and 30; in addition, rabies immunoglobulins (RIG) must be given to serious cases of exposure (grade III severity). Some studies indicate that these immunoglobulins suppress the immunogenicity of rabies vaccine when administered according to an alternative protocol of four injections (2-1-1) on days 0, 7, and 21, which was therefore not recommended for grade III exposures. To test this effect, we conducted a multicentre study in Indonesia using three groups of subjects. One g...
Hyperglobulinemia and lymphocyte subset changes in naturally infected, inapparent carriers of equine infectious anemia virus. To determine blood protein concentration, immunoglobulin concentration, and lymphocyte profiles in equine infectious anemia virus (EIAV) seropositive, naturally infected horses without clinical signs of disease. Methods: 26 clinically normal seropositive horses, 6 febrile ponies with experimentally induced EIA, and 52 clinically normal seronegative horses and ponies. Methods: Serum and EDTA-anticoagulated blood were obtained from all horses and ponies, and total serum protein and albumin concentrations, immunoglobulin concentrations, and blood lymphocyte subset counts were determined. Results:...
Organization of the equine immunoglobulin heavy chain constant region genes; III. Alignment of c mu, c gamma, c epsilon and c alpha genes. Previous restriction analysis of cloned equine DNA and genomic DNA of equine peripheral blood mononuclear cells had indicated the existence of one c epsilon, one c alpha and up to six c gamma genes in the haploid equine genome. The c epsilon and c alpha genes have been aligned on a 30 kb DNA fragment in the order 5' c epsilon-c alpha 3'. Here we describe the alignment of the equine c mu and c gamma genes by deletion analysis of one IgM, four IgG and two equine light chain expressing heterohybridomas. This analysis establishes the existence of six c gamma genes per haploid genome. The genomic a...
Immunologic function in horses after non-specific immunostimulant administration. Inactivated Propionibacterium acnes is a biologic response modifier for treatment of non-specific respiratory disease in horses. The objectives of this investigation were to determine alterations in phagocytic activity, phenotypic expression of lymphocyte subpopulations and lymphokine-activated killing cell response in healthy young horses. Samples were collected on day 0, 7 and 14 of the investigation. Blood samples were obtained via jugular venipuncture and pulmonary leukocytes were recovered via bronchoalveolar lavage (BAL). Commercially available P. acnes (Eqstim) was administered intraven...
Report of the Second Equine Leucocyte Antigen Workshop, Squaw valley, California, July 1995. The final assignment of antibody clusters for leucocyte antigens and immunoglobulins, as described in detail in Sections 3 and 4, is summarized in Table 4. Together with other mAbs developed outside of ELAW II (Table 9) this pool of reagents represent a powerful array of tools for the study of equine immunity. The Second Equine Leucocyte Antigen Workshop made considerable advances in pursuing the objectives of establishing the specificities of mAbs and achieving consensus on the nomenclature for equine leucocyte and immunoglobulin molecules. Of equal importance, several productive collaboratio...
Organization of the equine immunoglobulin constant heavy chain genes. I. c epsilon and c alpha genes. We provide a restriction map of the equine c epsilon and c alpha genes as a molecular basis for isotype classification. Human and murine DNA probes were used for identification of homologous equine DNA sequences and for isolation of the equine c epsilon and c alpha genes from a genomic DNA library. A detailed map of the equine 5'-s epsilon/c epsilon-s alpha/c alpha-3' gene region was obtained. Equine c epsilon and c alpha DNA probes were prepared and used for restriction analysis of immunoglobulin heavy chain gene loci from different horses. This analysis indicated the presence of only one equ...
Isolation and characterisation of equine dendritic cells. Despite their important role in initiating T-cell responses in other species, dendritic cells have not been studied in the horse. A method for isolating blood dendritic cells by adherence and metrizamide gradients was adapted to equine cells. A number of monoclonal antibodies (mAbs), including some which label dendritic cells in other species, were tested for immunochemical reactivity with the isolated blood dendritic cells, and sections of lymph node and spleen. 62 +/- 6% of the isolated blood cells were MHC Class II positive and had typical dendritic cell morphology and only 4 +/- 2% contain...
[Demonstration of immunoglobulin isotypes in the vitreous body as a contribution to the etiology of recurrent equine uveitis]. The functional properties of different immunoglobulin isotypes in equine recurrent uveitis (ERU) has not been investigated yet. Here, we describe the quantitative determination of total immunoglobulin levels and isotype differentiation in the vitreous of four horses with ERU as compared to that of seven healthy horses. In contrast to almost equal amounts of total immunoglobulin in the vitreous of both groups, remarkable differences were found: All four of the horses with ERU had significantly higher IgA contents in their vitreous as compared to the control group. However, the other isotypes mo...
Neonatal alloimmune thrombocytopenia in a quarter horse foal. Neonatal alloimmune thrombocytopenia is recognized as a spontaneous disease of human infants, piglets, and possibly mules, but it has not been previously reported in horses. A 1-day-old Quarter Horse foal presented to Michigan State University Large Animal Clinic with severe thrombocytopenia of unknown origin. Immunoglobulins that bound to the foal's platelets were identified in the mare's plasma, serum, and milk by indirect assays. The immunoglobulins were further shown to recognize platelets from the foal's full brother, born 1 year earlier. These findings, coupled with the clinical course o...
Expression cloning and humoral immune response to the nucleocapsid and membrane proteins of equine arteritis virus. To provide a convenient and sensitive method for the detection of equine arteritis virus (EAV)-specific serum antibodies, we developed an immunoblot assay employing the EAV nucleocapsid (N) and membrane (M) proteins expressed in a procaryotic expression vector (pMAL-c2) for the production of recombinant maltose-binding (MBP) fusion proteins (MBP-N and MBP-M). The antigenic reactivity of the recombinant fusion proteins and their Xa factor cleavage EAV products was confirmed by immunoblot using horse antisera to EAV. Some horse sera, however, showed immune reactivity to the MBP fusion partner pr...
Total protein and immunoglobulin concentrations in equine tears. Lacrimal fluid represents a major ocular surface defensive mechanism providing different concentrations of all immunoglobulin classes. In this report, four classes of immunoglobulins (IgA, IgM, IgG and IgGT) have been measured in horse tears. As in others species, IgA is the main immunoglobulin responsible for local protection and constitutes quantitatively, 50% of all lacrimal proteins. The rest of immunoglobulins studied are normally present in equine tear fluid (though in lower concentration) and contribute to ocular surface immune protection. Female and adult horses showed significant high...
Antigenic analysis of Rhodococcus equi preparations using different horse sera. An R. equi vaccine, prepared under conditions which induce the expression of many antigens, and which has given encouraging results in field trials, was analyzed by SDS-PAGE and immunoblots and compared with other R. equi preparations: a preparation made in with the same technique from a nonvirulent isolate (virulence associated protein negative, VapA-negative); a whole cell preparation of a VapA-positive R. equi, prepared as a standard bacterin; and a semipurified VapA preparation (APTX). The antigens in these preparations were analyzed using hyperimmune sera (from adult horses vaccinated wit...
Enzyme immunoassay for measuring 25-hydroxyvitamin D3 in serum. We developed a rapid, competitive enzyme immunoassay (EIA) for measuring 25-hydroxyvitamin D3 [25(OH)D3] in serum. The EIA was based upon 25(OH)D3-3-hemisuccinate covalently coupled to secondary amino groups grafted onto the polystyrene surface of microtiter wells. Optimal coupling conditions were established, and we found that inclusion of 40 mumol/L chloramine T, an agent not previously described for use in coupling to these plates, resulted in both more reproducible coupling as well as more than a twofold increase in the coupling efficiency. Before EIA, 25(OH)D3 was extracted from the serum...
Control of equine infectious anemia virus is not dependent on ADCC mediating antibodies. Horses infected with equine infectious anemia virus (EIAV) have recurrent episodes of viremia which are eventually controlled, but the immune mechanisms have not been identified. Antibodies were detected to the surface of EIAV-infected cells within 1 month postinfection and remained for at least 3.5 years postinfection. These antibodies recognized cell surface-exposed envelope (Env) glycoproteins, but could not mediate antibody dependent cellular cytotoxicity (ADCC) using EIAV-WSU5-infected equine kidney (EK) cells as targets and peripheral blood mononuclear cells (PBMC) or polymorphonuclear c...
Monoclonal gammopathy in a Dutch warmblood mare. A 15-year-old Dutch warmblood mare was presented because of lethargy, which had been present for several weeks, and severe anaemia. Total protein was high and serum electrophoresis revealed a monoclonal peak in the alpha-2 region. Monoclonal immunoglobulin, IgG(T), was detected by immuno-electrophoresis in serum and urine. Postmortem examination revealed a relatively large number of plasmacytoid cells in the bone marrow and a monotonous population of plasmacytoid cells in the spleen. These findings are suggestive of a plasma cell myeloma.
Failure of passive transfer in foals: incidence and outcome on four studs in New South Wales. To determine the regional incidence and effectiveness of treatment of failure of passive transfer (FPT) in foals. Methods: A study of disease incidence. Methods: Eighty-eight foals and 57 mares from four studs in the practice area of the Rural Veterinary Centre were tested. Methods: Foals were tested for their serum IgG and total serum protein (TSP) concentration within the first 72 hours of life. Colostrum was collected from mares and specific gravity determined. FPT and partial failure of passive transfer (PFPT) of immunoglobulins was diagnosed when serum IgG concentrations were < 4 g/L and ...
Characterization of horse (Equus caballus) immunoglobulin mu chain-encoding genes. Horse (Equus caballus) immunoglobulin mu chain-encoding (IgM) variable, joining, and constant gene segments were cloned and characterized. Nucleotide sequence analyses of 15 cDNA clones from a mesenteric lymph node library identified 7 unique variable gene segments, 5 separate joining segments, and a single constant region. Based on comparison with human sequences, horse variable segments could be grouped into either family 1 of immunoglobulin (Ig) clan I or family 4 of Ig clan II subclan IV. All horse sequences had a relatively conserved 16 base pair (bp) segment in framework 3 which was reco...
Immunohistochemical examination of light-chain expression (lambda/kappa ratio) in canine, feline, equine, bovine and porcine plasma cells. Five species of domestic animals were examined immunohistochemically and the light-chain ratios of the immunoglobulins produced by plasma cells analysed. Forty dogs, 11 cats, 10 horses, 11 cattle and 14 pigs were tested using the sequential indirect immunoperoxidase and immunophosphatase methods. Tissues from the tonsils, spleen and cervical lymph nodes were analysed. It could be seen that the lambda/kappa ratio in dogs, cats, horses and cattle is largely dominated by the lambda chains (lambda/kappa ratio in dogs: 91/9%, in cats; 92/8%; in horses: 96/4%; in cattle: 91/9%). A more or less balan...
The incidence and consequences of failure of passive transfer of immunity on a thoroughbred breeding farm. Circulating IgG concentration was determined between 12 and 24 hours after birth for 323 foals born on a Thoroughbred breeding farm over 3 consecutive years. The incidence of failure of passive transfer (FPT) of maternal immunoglobulins (foal circulating IgG concentration < 8 g/L) was found to be 9.6%. Foals born late in the season (October to December) were found to be at increased risk for the development of FPT. The degree of assistance required at parturition and the presence of a periparturient problem in the mare or foal also significantly influenced the subsequent incidence of FPT. Pass...
Antibody-mediated neutralization and binding-reversal studies on alpha-neurotoxins from Micrurus nigrocinctus nigrocinctus (coral snake) venom. An ELISA based, non-radioactive acetylcholine receptor (AchR) binding assay was used to detect the alpha-neurotoxins present in Micrurus nigrocinctus nigrocinctus venom. Sera from horses hyperimmunized against M. nigrocinctus venom contain antibodies which inhibit the binding of M. n. nigrocinctus alpha-neurotoxins to AchR and reverse the binding of toxins already complexed with the receptor. This result supports the importance of using antivenom therapeutically in M. n. nigrocinctus envenomations even after the onset of neurological symptoms. M. nigrocinctus antivenoms cross-reacted in an ELI...
Lymphoid leukosis viruses, their recognition as ‘persistent’ viruses and comparisons with certain other retroviruses of veterinary importance. Diseases caused by lymphoid leukosis virus (LLV), a retrovirus, take a long time after infection to develop and have a wide variety of pathological manifestations. This long latent period is characteristic of 'persistent virus infections'. Disease produced by LLV infection and its underlying mechanisms is compared with 'persistent' infections caused by other retroviruses in birds and mammals of veterinary importance. The diseases considered for comparison are those caused by reticuloendotheliosis, feline leukaemia, bovine leukosis and equine infectious anaemia viruses. There are significant ch...
Equine gammaherpesvirus 2 (EHV2) is latent in B lymphocytes. Peripheral blood leukocytes were collected from 5 Thoroughbred horses and examined for the presence of EHV2 in sub-populations of mononuclear cells. Peripheral blood mononuclear cells were separated on Percoll gradients and then enriched for plastic adherent cells (predominantly monocytes), surface immunoglobulin positive (sIg+) B lymphocytes and T lymphocytes, using panning techniques. The purity of each cell population was assessed by fluorescence activated cell scanning. In an infectious centre assay, each cell population was inoculated onto equine foetal kidney monolayer cell cultures whic...
Value of skin testing for predicting reactions to equine rabies immune globulin.
Clinical infectious diseases : an official publication of the Infectious Diseases Society of America
The high cost of postexposure prophylaxis for rabies is one reason that treatment is inadequate in developing countries. This problem has kindled interest in the use of equine rabies immune globulin, which is a less expensive, yet effective, substitute for human rabies immune globulin. Fatal anaphylaxis is a feared complication of the administration of heterologous serum; therefore, authoritative sources recommend prior skin testing. However, recommendations for methods of administering such a skin test and for its interpretation vary greatly. We embarked on a long-term study to develop guidel... Monoclonal equine IgM and IgG immunoglobulins. In order to define equine immunoglobulins (Igs) and to produce monoclonal reference Igs we fused equine peripheral blood mononuclear cells with X63-Ag8.653 non Ig producing murine myeloma cells. A total of 29 equine Ig producing equi-murine heterohybridomas were obtained, of which ten expressed equine Ig for more than 3 months. One of these heterohybridoma lines produced monoclonal IgM, an equine isotype which has not been available in monoclonal form before. Four lines secreted equine IgG of two distinct Ig heavy chain types as assessed by the molecular weight (MW), while the remaining five l...
Production of monoclonal antibodies in horses. Monoclonal antibodies (MAbs) have been successfully used to evaluate immune responses in horses, and to target important antigens of equine infectious agents to which protective immune responses may be directed (1–5). Most of these studies are performed with murine MAb produced by fusing spleen cells from immunized mice with an appropriate myeloma cell line, as described in Chapter 3. However, there are experimental questions for which murine MAb are not adequate. These include:
1.Definition of microbial epitopes recognized by the infected host;
2.Identification of immunodominant epito...
Seroanalysis of Tyzzer’s disease in horses: implications that multiple strains can infect Equidae. A monoclonal antibody based competitive inhibition assay was used to detect antibodies in horse sera to purified flagellar antigens from distinct Clostridium piliforme isolates. Sequential absorption of hyperimmune rat serum to C. piliforme isolate E (horse-origin isolate), a positive C. piliforme-immune horse serum, and other suspected immune horse sera with unrelated bacteria or C. piliforme isolates E or isolate R1 (rat-origin isolate) alone demonstrated the specificity of this assay for C. piliforme. This specificity was associated with the inhibition of monoclonal antibody binding to C. p...
The amino Acid sequence glutamine-628 to valine-646 within the A1 repeat domain mediates binding of von Willebrand factor to bovine brain sulfatides and equine tendon collagen. von Willebrand Factor (vWF) is a multifunctional glycoprotein in plasma and vascular subendothelial matrix which plays a major role in cellular adhesion. vWFdependent adhesion of platelets to the subendothelium at high shear rates involves a specific platelet membrane receptor, the glycoprotein (GP) Ib-IX complex. We have previously purified a 39/34-kiloDalton (kDa) dispase fragment of vWF (Leu-480/Val-481 to Gly-718) and demonstrated that this fragment contains the binding site for the GP Ib-IX complex [Andrews R K, et al. Biochemistry 1989; 28: 8326-83361. vWF also mediates agglutination of ...
Light chain isotype regulation in the horse. Characterization of Ig kappa genes. Horse Ig kappa genes have been characterized to determine whether there may be a structural basis for the low level of kappa expression in this species. The overall organization of the J kappa-C kappa locus is remarkably similar to that of the mouse and human loci. A single C kappa exon is separated by 2.9 kb from five J kappa segments, four of which seem functional and three of which are associated with canonical recombination signal sequences. A highly conserved intron enhancer was identified upstream of the C kappa exon and a single restriction fragment in horse genomic DNA hybridized stron...