Colostral transfer of bovine immunoglobulin E and dynamics of serum IgE in calves

The role of IgE in protective immunity is becoming understood, therefore the colostral transfer of IgE and the age-dependent changes of IgE levels may be important for neonatal immunity. To investigate this question, serum samples were collected from range-fed Hereford cows and their calves from birth through 9 months of age. The sera were assayed for total IgE by enzyme-linked immunosorbent assay (ELISA). Calves were found to have significant levels of IgE during the first week postpartum, indicating colostral transfer of IgE. Thereafter, serum levels declined rapidly within 3 weeks from birth. The IgE levels began to increase after 12 weeks of age, and in some cases reached adult levels. The passive transfer of maternal IgE through colostrum may be important in providing early protection from disease, especially against intestinal parasites.     

Quantitation and localization of immunoglobulin E containing cells in bovine lymphoid tissues

Although recent studies have begun to describe and quantify IgE responses in bovine serum and secretions, little is known about the distribution and quantity of IgE containing cells in cattle. In the present study, cells with cytoplasmic IgE were quantitated in bovine lymphoid tissues, using immunoperoxidase staining and evaluation by an image analysing computer (Quantimet). Frozen sections from retropharyngeal, bronchial and mesenteric lymph nodes, tonsil and spleen were stained from 11 calves, some of which had been exposed to antigen by aerosol or injection. Although individual variability was considerable, bronchial and mesenteric lymph nodes generally contained the greatest percentage of IgE containing cells, while retropharyngeal lymph node, tonsil, and spleen had less. Parenteral immunization with ovalbumin appeared to increase the splenic percentage, while aerosol exposure to ovalbumin was associated with a greater percentage of IgE containing cells in bronchial lymph nodes. Comparison of the present results with those reported for other species shows some similar trends in IgE localization.     

Demonstration of antibodies against bovine leukemia virus (BLV) by blocking ELISA using bovine polyclonal anti-BLV immunoglobulin.

A blocking enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against bovine leukemia virus (BLV) is described. The test is based on the biotin-streptavidin system using unlabelled polyclonal bovine IgG against BLV as catching antibody and biotinylated bovine anti-BLV IgG as detecting antibody. The sensitivity was found to be 50-100 times higher than the agar gel immunodiffusion test, with a specificity of practically 100%. The blocking ELISA proved to be suitable for detection of antibodies against BLV in serum and milk. In 34 paired milk/serum samples, the average ratio of BLV antibody titres was 1:26. So far, more than 700,000 sera have been screened by blocking ELISA for BLV antibodies in the course of the Danish surveillance programme for BLV infection.     

Generation and characterization of murine monoclonal antibodies specific for bovine immunoglobulin E

Monoclonal antibodies were produced against serum-derived bovine immunoglobulin E (IgE). Culture supernatants of hybridomas were initially screened by enzyme-linked immunosorbent assay (ELISA). Supernatant-derived antibodies were concentrated and further characterized using ELISA, reverse cutaneous anaphylaxis, immunohistochemical staining, and immunoblotting of IgE-containing samples separated by SDS-polyacrylamide gel electrophoresis (PAGE). Eight monoclonal antibodies showed specificity for bovine epsilon immunoglobulin heavy chain. Two antibodies (E2 and E32) reacted in immunoblots of SDS-PAGE of serum IgE under reducing conditions. Additionally, E2, E5, and E32 detected epsilon chain in serum separated by SDS-PAGE and then renatured. Antigen-specific IgE was detected in Western blots by E5 and E32. Immunoperoxidase staining of IgE-containing cells in mesenteric lymph node sections was detected with E5, E21 and E32. All eight antibodies produced positive reverse cutaneous anaphylaxis reactions in calf skin. All functioned well in ELISA as a plate-sensitizing reagent for quantitation of total IgE; E5 and E32 worked well as a primary antibody in antigen-specific IgE assays. These antibodies will be useful in research applications and in diagnostic assays.     

Studies of serum total immunoglobulin E concentrations in atopic and non-atopic dogs

The serum total immunoglobulin E (IgE) concentrations of two groups of atopic dogs and three groups of non-atopic dogs were compared. There was a wide range of concentrations with a high degree of overlap between the groups. The serum total IgE concentrations of a group of 15 non-atopic racing greyhounds were significantly higher than those of all the other groups. Atopic and non-atopic dogs receiving stringent parasite control treatments could not be differentiated on the basis of their serum total IgE concentrations. In the non-atopic dogs there was no correlation between their serum total IgE concentrations and the number of allergen-specific positive results obtained in an ELISA, or between their serum total IgE concentrations and their age.     

High allergen-specific serum immunoglobulin E levels in nonatopic West Highland white terriers

Human and canine atopic dermatitis (AD) share an association with IgE specific to environmental allergens, but few studies have evaluated serum allergen‐specific IgE in nonatopic dogs. This study compared serum allergen‐specific IgE levels in 30 atopic and 18 nonatopic West Highland white terriers. Atopic dermatitis was confirmed using standard criteria. Nonatopic dogs were over 5 years of age and had no clinical signs or history of AD. Serum allergen‐specific IgE levels were measured with Allercept^® IgE ELISAs using a 48‐allergen Australian panel. Positive reactions were defined as ≥150 ELISA absorbance units. Intradermal tests were performed in 16 atopic dogs, either at the time of or at various times prior to serum collection. In atopic dogs, the most common positive ELISA and intradermal test results were to Dermatophagoides farinae (11 of 30 dogs), but there were no statistically significant correlations between results from the two methods for any allergen. In nonatopic dogs, multiple high‐positive ELISA reactions were reported to 45 of 48 allergens, most commonly D. farinae and Tyrophagus putrescentiae (17 of 18 dogs each). Positive ELISA results in nonatopic dogs were statistically significantly higher than those in atopic dogs for 44 of 48 allergens, including two allergens (D. farinae and Dermatophagoides pteronyssinus) commonly regarded as significant in canine AD. In conclusion, positive allergen‐specific IgE ELISAs were not specific for canine AD, and high allergen‐specific IgE levels were seen in nonatopic dogs. The clinical significance of this and whether it characterizes a protective phenotype is unclear.     

Effect of vaccination on serum concentrations of total and antigen-specific immunoglobulin E in dogs

OBJECTIVE: To determine the effect of vaccination on serum concentrations of total and antigen-specific IgE in dogs. ANIMALS: 20 female Beagles. PROCEDURE: Groups of 5 dogs each were vaccinated repeatedly between 8 weeks and 4 years of age with a multivalent and rabies vaccine, a multivalent vaccine only, or a rabies vaccine only. A fourth group of 5 dogs served as unvaccinated controls. Serum concentrations of total immunoglobulins and antigen-specific IgE were determined following vaccination. RESULTS:-The multivalent vaccine had little effect on serum total IgE concentrations. The concentration of IgE increased slightly following vaccination for rabies at 16 weeks and 1 year of age and increased greatly after vaccination at 2 and 3 years of age in most dogs, with a distinct variation between individual dogs. Vaccination had no effect on serum concentrations of IgA, IgG, and IgM as measured at 2 and 3 years of age. The rabies vaccine contained aluminum adjuvant in contrast to the multivalent vaccine. An increase of IgE that was reactive with vaccine antigens, including bovine serum albumin and bovine fibronectin, was detected in some of the dogs vaccinated for rabies. There was no significant correlation between serum concentrations of total IgE and antigen-specific IgE following vaccination. Serum total IgE concentration rapidly returned to preimmunization concentrations in most dogs, but high concentrations of antigen-specific IgE persisted. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination of dogs for rabies increases serum concentrations of total IgE and induces IgE specific for vaccine antigens, including tissue culture residues. Vaccination history should be considered in the interpretation of serum total IgE concentrations.     

Hypersensitivity to dietary components in young farm animals: isolation and partial purification of bovine immunoglobulin E

An antibody of known specificity and active in long (72 hours) latent period passive cutaneous anaphylactic reactions, was isolated and partially purified from bovine serum. This antibody was not associated with immunoglobulins IgG, IgM or IgA. A rabbit antiserum raised against this antibody and used as an immunoabsorbent, successfully recovered skin sensitising antibody from bovine reaginic serum.     

Demonstration and quantitation of immunoglobulins in bovine serum, follicular fluid, and uterine and vaginal secretions with reference to bovine viral diarrhea and infectious bovine rhinotracheitis.

Immunoglobulin concentrations (IgG, IgM, and IgA) in bovine serum, follicular fluid, and uterine and vaginal secretions were determined. The specificities of IgG, IgM, and IgA for virus-neutralizing antibody against bovine viral diarrhea (BVD) and infectious bovine rhinotracheitis (IBR) viruses were also examined. High concentrations of IgG were present in both serum and follicular fluid. The IgG, IgM, and IgA concentrations were low in uterine and vaginal secretions. There was more IgG in the uterus during estrus than at any other time. Virus-neutralizing antibodies against BVD and IBR in serum of cows were mainly the IgG class. There was positive correlation between serum and follicular fluid virus-neutralizing antibody titers fro BVD and IBR. These antibodies may provide some protection for recently ovulated ova.     

Comparison of the 2-mercaptoethanol and dithiothreitol tests for determining Brucella immunoglobulin G agglutinating antibody in bovine serum.

The dithiothreitol test was evaluated as a substitute for the 2-mercaptoethanol test for determining Brucella immunoglobulin G agglutinating antibody in bovine serum. The tests were compared on 207 card-positive sera that showed a standard tube-agglutination titer of incomplete 1:50 or higher. The tests agreed within one dilution with 182 of the 207 sera tested for an 87.9% rate of agreement. When titers were not the same, those obtained with the dithiothreitol test were more frequently lower than higher than those obtained with the 2-mercaptoethanol test. Sixteen sera that showed a titer with the 2-mercaptoethanol test were negative with the dithiothreitol test and two that showed a titer with the dithiothreitol test were negative with the 2-mercaptoethanol test. Results suggest that the dithiothreitol test is not a reliable substitute for the 2-mercaptoethanol test to detect immunoglobulin G agglutinating antibody in bovine serum.     

A study of bovine and equine immunoglobulin levels in pony foals fed bovine colostrum

As part of a project to raise specific pathogen free (SPF) Welsh Mountain Pony foals, free from exposure to Equid herpesvirus type 1, foals were removed from their dams at birth and fed bovine colostrum. This study characterises the uptake of bovine colostral immunoglobulin and production of endogenous immunoglobulin, in 10 SPF foals. An enzyme-linked immunoadsorbent assay was developed to measure serum concentrations of bovine IgG1 (boIgG1) to assess the efficiency of transfer, and rate of elimination of boIgG1 by the foal. The endogenous production of equine IgG was studied using a single radial immunodiffusion test. Foals were given 1.2 to 2 litres of bovine colostrum achieving peak serum boIgG1 concentrations of 18.9 to 34.2 g/litre (mean 28.0). The mean half-life of boIgG1 in the foals was 7.4 days. Endogenous immunoglobulin production resulted in equine IgG concentrations greater than 2 g/litre in six of 10 foals by 14 to 19 days of age, and greater than 7 g/litre in eight of 10 foals by 37 to 50 days of age. All foals had equine IgG serum concentrations greater than 10 g/litre by 102 to 135 days of age.     

Demonstration of infectious bovine rhinotracheitis virus antigen in paraffin sections

Nine pregnant heifers were inoculated intravenously with infectious bovine rhinotracheitis virus (IBRV) in the sixth month of pregnancy. Tissues were collected from the fetus of a heifer killed 13 days postinoculation (PI), from fetuses of 6 heifers that aborted 16-27 days PI, and from mummified fetuses of 2 heifers that aborted 53 and 85 days PI, respectively. Control tissues were obtained from the fetus of a non-inoculated heifer that was killed in the seventh month of gestation. Tissues were fixed in 10% formalin, embedded in paraffin, and examined for viral antigen by immunohistochemistry, using biotinylated second antibody and alkaline phosphatase-labeled avidin-biotin complex. Antigen was detected in at least 1 tissue from the fetus of each inoculated heifer. Positive tissues included lung, liver, spleen, kidney, adrenal, and placenta. In several fetuses, antigen was identified in tissues from which virus was not isolated in cell culture. This appeared to occur when tissues had only a few small foci of infection or when tissues were severely autolyzed. The observation of viral antigen in tissues from mummified fetuses indicates that this technique may be useful in diagnostic laboratories to detect IBRV infection in tissues that are not suitable for virus isolation or for examination by the cryostat tissue section-fluorescent antibody technique.     

Structure and diversification of the bovine immunoglobulin repertoire

Our understanding of the basis to immunoglobulin formation in cattle has benefited substantially from the application of molecular biology over the past decade. It is now established that both the lambda light chain and heavy chain repertoires are founded upon the frequent expression of single gene families and subgroups of segments which are of conserved sequence. It is likely that a functional kappa locus exists in the bovine genome but this isotype comprises as few as 5% of bovine light chains. Similarly, alternative but non-expressed V(H) gene families are present posing intriguing but unresolved questions about the regulation of immunoglobulin synthesis. The heavy chain frequently bears a third complementarity-determining region which is atypically long but the processes which expand this region of the reading frame and its contribution to the interaction with antigen remain matters of speculation. Opportunities exist to map the major immunoglobulin loci and to define the membership and sequence diversity of the gene families which dominate each repertoire. However, it is already evident that cattle cannot generate significant diversity from rearrangement and junctional imprecision alone. Elucidation of the mechanism(s), dynamics and tissue distribution of immunoglobulin diversification in cattle, thus, remain key challenges in this branch of veterinary immunology.     

Involvement of calcium in mitogenic and immunoglobulin responses in bovine lymphocytes

Removal of calcium from cultures of bovine splenic lymphoid cells with ethyleneglycol-bis-(2-amino ethylether)-N,N'-tetraacetic acid (EGTA) caused significant suppression of pokeweed mitogen-induced immunoglobulin (Ig) biosynthesis. It was found that removal of calcium for 48 hours of a 5-day culture significantly altered Ig biosynthesis. Re-addition of calcium after 48 hours did not restore Ig synthesis. The EGTA caused significant (P < 0.05) suppression of DNA synthesis when calcium was removed from concanavalin A- and purified protein derivative-stimulated peripheral blood lymphocyte cultures. Further, removal of calcium for 48 hours of 4-day cultures significantly altered the DNA synthesis in mitogen- and antigen-stimulated cultures.     

Quantitation of bovine macrophage colony-stimulating factor in bovine serum by ELISA

We established an enzyme-linked immunosorbent assay (ELISA) system for the quantitation of bovine macrophage colony-stimulating factor (M-CSF) and used it to measure the serum M-CSF levels in bovine fetuses and calves. The average serum M-CSF level was 2.7+/-1.5 ng/ml in 39 calves under 100 days old, and 1.8+/-0.8 ng/ml in 15 cattle between 101 and 418 days old. Fetal sera samples (n = 6) prepared from cattle between 150 and 280 days of gestational age had a higher average level of M-CSF (8.8+/-1.4 ng/ml). Alteration in serum M-CSF levels in each individual calf was also measured. The serum levels of M-CSF in calves at 0-1 day after birth ranged from 0.52 to 7.3 ng/ml. During the period 113-125 days after birth, serum levels were around 1.4+/-0.39 ng/ml. Although serum M-CSF levels generally decreased as the age of calves advanced, differences among individuals, especially among newborn calves, were observed.