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Topic:Immunoglobulin G
Immunoglobulin G (IgG) is the predominant antibody isotype found in the bloodstream of horses and is integral to their immune defense mechanisms. It is produced by B lymphocytes and plays a significant role in identifying and neutralizing pathogens such as bacteria and viruses. IgG is involved in various immune functions, including opsonization, complement activation, and neutralization of toxins. In equine medicine, measuring IgG levels is important for assessing the immune status of foals, especially in cases of failure of passive transfer (FPT) of maternal antibodies. This page compiles peer-reviewed research studies and scholarly articles that explore the structure, function, and clinical implications of Immunoglobulin G in equine health.
Separation of equine IgG subclasses (IgGa, IgGb and IgG(T)) using their differential binding characteristics for staphylococcal protein A and streptococcal protein G. Equine IgG possesses four well-defined subisotypes, designated IgGa, IgGb, IgGc and IgG(T) on the basis of their increasing anodal mobility in electrophoresis. However, the preparation of IgGa and IgGb reference proteins has not previously been reported. Certain bacterial cell wall proteins, termed protein A and protein G, have been used for purification of IgG subisotypes from the serum of domestic animals which, combined with other techniques utilising differences in the physico-chemical properties of the proteins, has allowed the purification of Ig isotypes. This paper describes purificatio...
Correlation of antigen specific IgG and IgG(T) responses with Anoplocephala perfoliata infection intensity in the horse. There is increasing interest in the application of serological methods to macro-parasite infections to indicate infection intensity, which in turn is related to pathogenicity. Colic is the single most important cause of mortality in horses and there is evidence that a proportion of colic cases are associated with infection with the intestinal cestode Anoplocephala perfoliata. In order to develop better tools to investigate this association, the correlation between antigen-specific equine IgG and IgG(T) and infection intensity of A. perfoliata was investigated. Affinity purification of a 12/13 ...
Serum antibody responses of foals to virulence-associated 15- to 17-kilodalton antigens of Rhodococcus equi. Humoral immune responses in 16 foals to virulence-associated 15- to 17-kDa antigens of Rhodococcus equi were studied during the first fourteen weeks of life on two horse-breeding farms with a persistent incidence of R. equi infection. Serum antibody levels specific for 15- to 17-kDa antigens were measured by enzyme-linked immunosorbent assay and Western immunoblotting. Immunoglobulin G (IgG) antibodies specific to 15- to 17-kDa antigens were detected by all the foals. R. equi was found in the feces of foals during week 1 of life, and the number of fecal R. equi rapidly increased to the highest...
Positive selection of EqCD8+ precursors increases equine lymphokine-activated killing. Lymphokine activated killing (LAK) is an example of natural cytotoxicity, and as such is a critical means of defense against diseases such as viral infection and neoplasia. Despite this important role, the specific molecular interactions involved in LAK or other forms of natural cytotoxicity are only partially understood. In some species, cells capable of mediating natural cytotoxicity express the CD8 molecule, although no specific role has been demonstrated for CD8 in non-MHC restricted cytotoxicity. In this study the role of the EqCD8 equine homolog of CD8 in LAK cell activity was examined. ...
Immunohistochemical analysis of an equine model of synovitis-induced arthritis. To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone. Methods: 2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2). Methods: Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 micrograms/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and a morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articula...
Generation of in vitro natural cytotoxicity of horse lymphocytes against sarcoid-derived tumor cells not expressing major histocompatibility complex antigens. To analyze in vitro lymphocyte-mediated immune responses of horses with sarcoids against allogeneic sarcoid cells containing endogenous retrovirus but not expressing major histocompatibility complex antigens. Methods: Lymphocyte-mediated immune reactions were assessed by means of proliferative responses in mixed lymphocyte tumor cell culture (MLTC) assay and lymphocyte-mediated cytotoxicity against various equine target cells. Methods: 12 horses with sarcoid tumors and 15 control horses. Methods: Blood lymphocytes were cocultured in MLTC with allogeneic sarcoid cells (Mc-1, BayMc-1), equine te...
Cross-antigenicity of horse serum albumin with dog and cat albumins: study of three short peptides with significant inhibitory activity towards specific human IgE and IgG antibodies. Horse serum albumin is present in the near vicinity of the animal, while dog and cat serum albumins are very common allergens present in house dust. Human patients clinically defined as allergic to horse could react with horse serum albumin by means of IgE or IgG antibodies. Studies regarding the specificities of these antibodies by inhibition enzyme-linked immunosorbent assay (ELISA) and depletion experiments have demonstrated that they are directed against dog serum albumin and cross-react not only with horse serum albumin but with other serum albumins from different origins. To investigate ...
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. P...
Effect of vaccination of ponies with A4 anti-idiotypic antibody on serum idiotype (1C9) and antilipid A concentration. To evaluate the humoral response of horses to vaccination, using a murine monoclonal anti-idiotypic antibody (A4) that shares an epitope with lipid A. Methods: Serum concentrations of antilipid A antibody and 1C9 (epitope on murine monoclonal antilipid A antibody) were measured serially during the period of vaccination with A4. Methods: 6 clinically normal adult ponies. Methods: Ponies were inoculated IM 3 times at monthly intervals with A4. Two weeks after each inoculation, serum was obtained and was assayed by ELISA for antilipid A and 1C9 concentrations. Additional vaccinations were given t...
Blastogenic response of lymphocytes from foals infected with Rhodococcus equi. The blastogenic response of lymphocytes from 16 newborn foals naturally infected with Rhodococcus equi was investigated, in order to evaluate the relationship between R. equi infection and depressed host response. Naturally infected foals showed evidence of R. equi infection at 5-6 weeks of age, as determined by clinical, haematological, bacteriological and serological methods. The blastogenic response of lymphocytes against phytohaemagglutinin was significantly depressed (stimulation index < 1.80; P < 0.01, P < 0.05) in R. equi-infected foals at 5-6 weeks of age compared with those o...
Histologic and immunohistochemical characterization of hemangiomas in the skin of seven young horses. The histologic and immunochemical characteristics of benign vascular tumors excised from the skin of seven young horses were evaluated. The patients were male horses of various breeds and were 16 months of age or younger at the time of presentation. Six tumors occurred on the extremities, and one was removed from the lip. Histologically, most tumors consisted of cellular nodules of varying compactness with few to many blood-filled lumina. Nodular growth separated preexisting adnexa and subcutaneous collagen. Some tumors contained or consisted predominantly of loosely packed arrays of ramifying...
Lack of acid-resistant trypsin inhibitor in mare’s colostrum: short communication. Mare's colostrum was collected and examined for the presence of trypsin inhibitors. It was found to contain a low level of trypsin inhibitor which could be denatured by 2.5% trichloroacetic acid and, therefore, it clearly differs from the acid-resistant colostral inhibitor of Artiodactyla and Carnivora. This finding is exceptional for a species that concentrates IgG in the colostrum and whose newborn absorbs colostral proteins non-selectively by the gut. It appears that the presence of colostral trypsin inhibitor is not essential for the transmission of maternal immunity via the colostrum and ...
Interleukin-1-mediated regulation of plasminogen activation in pregnant mare serum gonadotropin-primed rat granulosa cells is independent of prostaglandin production. This study examines the effects of interleukin-1 (IL-1) on plasminogen activator (PA) activity and prostaglandin (PG) E production in pregnant mare serum gonadotropin (PMSG)-primed granulosa cells and the potential involvement of PGE in the regulation of ovarian plasminogen activation. Methods: Granulosa cells were obtained from PMSG-primed rat (27-day-old) ovaries and cultured in serum-free conditions for 48 hours in the absence or presence of IL-1 beta (10 ng/mL) with and without transforming growth factor-beta 1 (10 ng/mL). Cellular PA activity was measured through the conversion of plasmin...
Monoclonal antibodies specific for equine IgG sub-isotypes including an antibody which recognizes B lymphocytes. Equine immunoglobulin G is currently classified as consisting of five sub-isotypes: IgGa, b, and c, IgG(T), and IgG(B). The study of the role of these immunoglobulins in antigen-specific responses, and the examination of their functional properties would be greatly facilitated by the availability of monoclonal antibodies (Mabs) that distinguish between them. The production and characterization of two Mabs that recognize an IgG sub-isotype with the characteristics of IgG(ab) is described. The immunoglobulin identified by these Mabs had a heavy chain weight of 53 kDa, was of rapid cathodal elect...
In vitro and in vivo evidence on the site of neutralization of equine chorionic gonadotrophin (eCG) by an eCG antiserum. This study was designed to determine whether the major site of eCG neutralization by an antiserum to the hormone is at the peripheral or ovarian level. Hamsters hypophysectomized at oestrus were injected s.c. with 25 iu eCG. Three days later, preovulatory follicles were dissected and cultured for 5 h and the medium was changed every hour. At the end of the first hour of incubation, oestradiol and androstenedione accumulation was high, with a sharp drop over the next 4 h, whereas progesterone concentrations did not change over the entire period. Addition of eCG antiserum to the incubated follic...
The equine endometrial cup reaction: a review. The function of eCG in equine pregnancy is far from clear but it has become evident that eCG has little or no FSH activity in the horse and is therefore probably not responsible for the secondary ovulations. eCG does have luteotrophic activity and it could play a role in the resurgence of the primary corpus luteum (1,7,44). Some evidence exists that the receptor population on the equine gonads is heterogenous in a way that makes it possible to distinguish eCG from eLH, resulting in different post-receptor effects (7). There is also evidence that eCG itself is heterogenous, both in glycosylatio...
An 80-kDa syncytiotrophoblast alloantigen bound to maternal alloantibody in term placenta. We have shown that most of the IgG present on term syncytiotrophoblast, membrane, microvesicles is bound to an 80 kDa protein antigen (R80K). Methods: Microvesicles were prepared from term human placenta, and the IgG eluted at pH3. Results: When IgG antibody was eluted at pH3 and reacted with acid-treated vesicles of other placentae, the alloantibody always bound to the preparation from which it was obtained, but only to about 10% of acid-treated preparations from other placentae. A similar polymorphic protein found in association with IgG antibody was found in term horse placentae. Cross-reac...
Cloning and expression of two genes from Babesia equi merozoites and evaluation of their diagnostic potential. High-titre equine immune sera were used to screen a lambda gt 11 expression library of Babesia equi cDNA fragments. Two cDNA clones which did not cross-hybridize to each other were studied. Both clones hybridized specifically to DNA from B. equi but not to DNA from B. caballi, B. divergens or B. ovis. Recombinant proteins were expressed as glutathione S-transferase (GST) fusion proteins with apparent molecular weights of 40 kDa and 75 kDa. Polyclonal antibodies directed against the 40 kDa and 75 kDa recombinant proteins detected native antigens of 55 kDa and 50 kDa respectively in crude lysate...
Localization of a protective epitope on a Venezuelan equine encephalomyelitis (VEE) virus peptide that protects mice from both epizootic and enzootic VEE virus challenge and is immunogenic in horses. In order to define more precisely the protective epitope encoded within the first 25 amino acids (aa) of the E2 glycoprotein of the Trinidad donkey strain of Venezuelan equine encephalomyelitis (VEE) virus, we examined the immunogenicity of smaller peptides within the first 19 aa. pep1-9 and pep3-10 elicited virus-reactive antibody, but failed to protect mice from virus challenge. Additionally, pep3-10 was identified by a competitive binding assay using overlapping peptide octamers as the putative binding site of the antipeptide monoclonal antibody (mAb) 1A2B-10. Since the E2 amino-terminal se...
Immunisation of mares to control endometritis caused by Streptococcus zooepidemicus. Normal mares were immunised by the intramuscular and intrauterine administration of an antigen with adjuvant and they and unimmunised control mares were later challenged by the intrauterine instillation of pathogenic Streptococcus zooepidemicus; the response of all the mares was monitored clinically and bacteriologically for seven days. Significantly fewer S zooepidemicus were present in cervical swabs taken from the immunised mares than from the control mares (P < 0.01) and the degree of inflammation in the genital tract of the immunised mares was also significantly less (P < 0.001). Th...
Experimental immunization of ponies with Strongylus vulgaris radiation-attenuated larvae or crude soluble somatic extracts from larval or adult stages. Protection from Strongylus vulgaris infection through immunization with radiation-attenuated third-stage larvae (L3) or crude soluble homogenates from larval or adult stages was examined. Yearling ponies raised parasite-free were divided into 3 immunization groups: radiation-attenuated L3; soluble adult somatic extracts; larval somatic extracts with excretory/secretory products (E/S) from in vitro culture; and 1 medium control group. Ponies were immunized twice; attenuated larvae were administered orally and somatic extracts or controls injected intramuscularly with adjuvant. Approximately 6 w...
Evaluation of a monoclonal antibody-based colony blot test for rapid identification of virulent Rhodococcus equi. We recently generated a monoclonal antibody immunoglobulin G1 (MAb 10G5), which can recognize 15- to 17-kDa antigens, virulence-associated antigens of Rhodococcus equi, and developed a colony blot enzyme-linked immunosorbent assay with MAb 10G5 for the rapid identification of virulent R. equi. In this epidemiologic study, we evaluated the results of the colony blot test in the identification of virulent isolates of R. equi from feces of horses and soil and compared them with those from a conventional procedure (plasmid profiles of isolates by agarose gel electrophoresis). Environmental isolate...
Inhibition of binding, entry, or intracellular proliferation of Ehrlichia risticii in P388D1 cells by anti-E. risticii serum, immunoglobulin G, or Fab fragment. The effects of equine antiserum, immunoglobulin G (IgG) specific for Ehrlichia risticii, and its Fab fragment on E. risticii binding to, internalization into, and proliferation in P388D1 cells were studied by immunofluorescence flow cytometry. Anti-E. risticii equine serum or IgG inhibited E. risticii at a stage beyond binding and internalization. In contrast, monovalent anti-E. risticii equine Fab fragments inhibited E. risticii binding and internalization into P388D1 cells. In the presence of control equine serum, IgG, or its Fab fragment, E. risticii cells were bound, were internalized and ...
Variation in expression of MHC class II antigens on horse lymphocytes determined by MHC haplotype. A panel of monoclonal antibodies was used to characterize the expression of equine Major Histocompatibility Complex (MHC) class II antigens on lymphocytes of horses of different MHC types. MHC class II antigen expression was compared between adult horses and foals, and the level of expression of MHC class II antigens on horse T cell subpopulations was also determined. Peripheral blood lymphocytes (PBL) from young and adult healthy horses of different MHC haplotypes were labeled with the antibodies and assayed by single- and two-color immunofluorescence flow cytometry. A variation in the expres...
Hemolytic complement activity and concentrations of its third component during maturation of the immune response in colostrum-deprived foals. Six foals were deprived of colostrum for the first 36 hours after birth and, instead, received reconstituted powdered milk. Five control foals suckled their dams naturally. Blood samples were obtained from all the foals after birth and at approximately weekly intervals until at least 5.5 months of age. Sera were analyzed for hemolytic complement activity, complement component C3, and correlating IgG concentration. Hemolytic complement (P = 0.0145) and C3 (P = 0.0002) values were significantly higher in colostrum-deprived foals (CDF) than in naturally nursed foals at 2 to 5 days of age. In addi...
Albumin quotient, IgG concentration, and IgG index determinations in cerebrospinal fluid of neonatal foals. Total protein (TP), albumin, and IgG concentrations were measured in CSF from the atlanto-occipital (AO) and lumbosacral (LS) sites and in serum of 15 clinically normal neonatal foals 85.2 mg/dl or AQ > 2.4, as determined by mean +/- 2 SD, may indicate increased blood-brain barrier permeability. An IgG index value > 1.0 may indicate intrathecal IgG production. Values obtained for foals of this study should serve as baseline for comparison in the evaluation of blood-brain barrier permeability and intrathecal IgG production in neonatal foals with neurologic disease.
Snake antivenoms from hyperimmunized horses: comparison of the antivenom activity and biological properties of their whole IgG and F(ab’)2 fragments. IgG and F(ab')2 fragments were prepared from horse plasma rich in specific antibodies against Brazilian Bothrops or Crotalus venoms. Both preparations, free of gross contamination with non-immunoglobulin proteins, were able to combine in vitro with their respective antigens, forming immune complexes at antigen excess, equivalence or antibody excess, and activating the C system, through either the classical or the alternative pathways. The IgG preparation was more effective in neutralizing the lethal factors in Bothrops or Crotalus venoms, compared with the F(ab')2 fragments. In contrast, IgG a...
Borrelia burgdorferi infection in UK horses. Antibody levels (IgG and IgM) to Borrelia burgdorferi were measured in the sera and synovial fluids of UK horses. Western blotting against B. burgdorferi was also used on samples from seropositive horses. A low incidence of seropositivity was shown in horses from most parts of the UK. This increased in areas that have a high incidence of human and canine borreliosis (Norfolk and south coast). Leptospira infections of horses did not cause cross reactions in the B. burgdorferi ELISA. Most horses did not display clinical signs of Lyme disease. As with dogs and man, it is apparent that B. burgdorf...
Influence of estrogen on antibacterial and immunoglobulin secretory activities of uterine fluids from ovariectomized mares. Effect of estrogen (E2) and progesterone (P4) on uterine antibacterial activity and immunoglobulin concentrations in mares was studied. In 2 in vitro experiments, 6 mixed-breed mares were ovariectomized, and uterine fluid and blood serum were analyzed. Antibacterial assay methods were used to determine inhibitory effects on Streptococcus zooepidemicus of uterine fluid samples collected on days 3, 5, and 8, and serum obtained on day 8 of treatment. Single radial immunodiffusion methods were used to quantify amounts of IgA and IgG in uterine fluid and serum on days 3, 5, 8, and 14 of treatment. ...
Identification of 2 stallion sperm-specific proteins and their autoantibody response. In this study, 2 stallions were immunised with their own spermatozoa to ascertain whether an antisperm autoantibody response could be mounted. The results demonstrated that the stallion can recognise and respond to sperm autoantigens by producing circulating antisperm antibodies, primarily of the IgG class. Such autoantibodies appeared 2-4 weeks after inoculation and persisted for 6-20 weeks. Immunochemical characterisation by western blot identified two major sperm autoantigens, with molecular weights of 70 kD and 62 kD. Control pony stallions immunised with adjuvants alone failed to exhibit ...
