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PubMed1986

Quantitative measurement of human immunoglobulin E using monoclonal antibodies to distinct epitopes.

Monoclonal antibodies (MAb) which recognize distinct epitopes on human immunoglobulin E have been used to develop two-site sandwich radio- and enzyme-linked immunoassays for the quantitation of human IgE. In the first step, a purified anti-IgE MAb coated to polyvinyl or polystyrene microtiter plates specifically bound the IgE contained in the samples. In the second step, another anti-IgE MAb (either iodinated or conjugated to beta-galactosidase) directed to a different antigenic determinant was used to estimate the amount of bound IgE. This simple method permitted the determination of IgE concentrations of 10 ng/ml and greater in about 3 h. Coefficients of variation on a single day did not exceed 7.5% for IgE levels, covering a wide range of the standard curve. The values obtained on serum samples showed a good correlation with those obtained using the paper radioimmunosorbent test (PRIST).

Antibodies, Anti-Idiotypic↗
PubMed1981

A U.S. reference for human immunoglobulin E.

The Committee on In Vitro Tests of the American Academy of Allergy has collected a large pool of human serum containing high titers of total IgE for the purpose of establishing a U.S. reference material. This pool is free from hepatitis B surface antigen. The serum pool has been subaliquoted, lyophilized and tested for total IgE content. The paper radioimmunosorbent test (PRIST) technique was used for this measurement. All laboratories used the same lot of reagents. The Second International Reference Preparation was used as the reference standard in each assay. Multiple replicates of the pooled sera was tested for total IgE content with one set of reagents by each of the 14 participating committee laboratories. The results for total IgE content (IU) of the vials were: mean of means, 899; median of means, 901; weighted mean, 898; with an overall coefficient of variation of 15.3%. This new IgE reference material is available to all laboratories for use as a primary reference standard from the Research Resources Branch, NIAID-NIH, Bethesda, MD 20205.

Humans↗
PubMed1985

IgE level in some dermatological diseases.

The total IgE level was measured by radioimmunosorbent test (Phadebas RIST, Pharmacia) in blood samples of 454 dermatological patients and 26 blood donors aged 14-70 years. The results showed different IgE values in the same groups of diseases. Elevated serum IgE values were observed in atopic dermatitis, drug allergy, discoid lupus erythematosus and chronic dermatomycosis. In drug allergy the average nonspecific IgE level of patients reacting with immediate reaction was higher than the normal value. Tested individually with Penicilloyl G RAST no strong correlation could be demonstrated between the quantity of specific and non-specific IgE.

Adolescent↗
PubMed1991

Detection of immunoglobulins from explant cultures of periapical lesions.

The presence of immunoreactive cells in periapical inflammatory lesions suggests that immune responses participate in the disease process. The purpose of this study was to determine the presence and concentration of immunoglobulins in the supernatant fluids of explant cultures of periapical lesions. Ninety periapical lesions that had been contiguous with the apex of a root were removed and maintained in explant cultures for 96 h. Tissue culture medium was replenished at 24, 48, 72, and 96 h. Double diffusion in agarose assays demonstrated the presence of IgG in 100% of the 24-h supernatant fluids and IgA in 65% of the 24-h supernatant fluids. However, IgM was not detected. Radial immunodiffusion assays were used to detect and quantitate IgG, IgA, and IgM in samples of 24-h supernatant fluids from 90 explant cultures. IgG was the predominant immunoglobulin followed by IgA. A radioimmunosorbent test was used to detect and quantitate IgE in samples of 24-h supernatant fluids from 90 explant cultures. Forty of the 90 supernatant fluids contained measurable IgE. All detected immunoglobulins decreased in concentration in daily supernatant fluids with time (24, 48, 72, and 96 h) in the culture.

Adolescent↗
PubMed1989

Nonspecific immunoglobulin E in aqueous humor: evaluation in uveitis.

The levels of immunoglobulin E (IgE) in aqueous humor (AH) and serum were determined in 74 uveitis patients and 13 control patients with senile cataract. Uveitis patients' levels ranged from less than 0.06 to 460 IU/ml according to the paper radioimmunosorbent test (PRIST; limit of sensitivity, 0.06 IU/ml). Only one control patient had an AH level higher than the sensitivity limit (0.08 IU/ml). Of the five patients with the highest levels in the AH, four had toxocaral retinitis. The ratio of AH/serum to IgE normalized to the albumin concentration was calculated in 25 uveitis patients and was greater than 0.60 in 13 cases, indicating a possible intraocular IgE synthesis.

Adolescent↗
PubMed1983

Lack of pokeweed mitogen-induced IgE formation in vitro by human peripheral blood mononuclear cells: detection of cross-reacting idiotypic determinants on polyclonal Ig by antibodies to a single IgE myeloma protein.

The ability of human peripheral blood mononuclear cells to synthesize IgE in vitro in response to pokeweed mitogen (PWM) is controversial. To determine whether the conflicting results obtained by different laboratories could be due to inherent qualitative differences in the anti-IgE antibodies used to measure low concentrations of IgE in culture supernatants, we compared the specificities of anti-IgE reagents prepared by various methods. Immunoadsorbent-purified antibodies were isolated from a goat antiserum to the lambda, IgE myeloma protein PS and a rabbit antiserum to the kappa, IgE protein Bed in three ways: 1) antibodies to IgE PS (anti-PS) were isolated from the goat antiserum by affinity chromatography with PS coupled to Sepharose 4B; these antibodies consisted of anti-epsilon chain-specific and anti-idiotypic antibodies to protein PS; 2) antibodies specific for the epsilon-chain (anti-epsilon) were purified by affinity chromatography with IgE myeloma proteins that were not used for immunization; and 3) antibodies to idiotypic determinants of proteins PS (anti-id PS) and Bed (anti-id Bed) were isolated on affinity columns with the respective myeloma proteins after absorption of the epsilon-chain-specific antibodies. These three types of antibodies were then used in a solid phase radioimmunosorbent test to quantitate the amount of "IgE" synthesized by peripheral blood mononuclear cells from nonatopic and atopic donors cultured for 7 days in the presence and absence of PWM. ANTI-PS antibodies detected a PWM-induced "IgE formation" in cell culture supernatants of both non-atopic and atopic donors. (ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Anti-Idiotypic↗
PubMed1976

Immunoglobulins in nasal secretion with special reference to IgE. I. Methodological studies.

Different ways of collecting relatively large volumes of nasal secretion with as physiological a composition as possible were studied. Nasal secretion was collected by the so-called nasal spray washing method from 5 patients with allergic rhinitis due to pollen and 5 healthy persons during a pollen-free season. The collection was performed without any provocation and following nasal provocation with histamine or allergen solution. With the radioimmunosorbent test, in which the lower limit of measurement was 0.1 units/ml, IgE could be quantified in 52 of 60 analysed secretions. IgA, IgG and albumin were demonstrated in all secretions. In the allergic patients, following histamine and allergen provocation, a relative increase in the concentration of IgE and albumin and a significant decrease of the IgA/albumin ratio in nasal secretion was found. In the healthy subjects, such changes in the secretion were observed only after histamine provocation. Calculations also suggested some local production of IgE, but not at all of the same order of magnitude as of IgA.

Adult↗
PubMed1979

Analysis of the serum IgE levels in nonimmunized rats of various strains by a radioimmunoassay.

An assay, the paper radioimmunosorbent test (PRIST) for the measurement of total serum IgE in the rat, that allows the measurement of IgE down to a level of 0.25 ng/ml, is described. With this assay it has been demonstrated that significant differences exist in normal serum IgE levels in non-immunized rats. These differences are strain-dependent, and strains may be classified as to low (less than 50 ng/ml), medium (50--300 ng/ml) or high (greater than 300 ng/ml) serum levels. The serum IgE level is not sex-dependent. Environmental factors have a pronounced influence, optimal housing conditions resulting in low and stable levels while suboptimal conditions result in high levels with great fluctuations. Hybridization experiments suggest a multigenic control, with low IgE strains having a dominating suppressor effect.

Age Factors↗
PubMedDate not supplied

Radioimmunoassay of IgE in middle ear effusions.

Fifty-six sample pairs of middle ear effusions and sera obtained from patients with otitis media with effusion were investigated for The IgE antibody activity against mites, using radioallergosorbent test (RAST) which was recently developed as an in vitro allergy test. The IgE concentrations were also determined by radioimmunosorbent test (RIST). Results showed that the IgE antibody activity against this allergen was found in 5 middle ear effusions and 3 sera. Four of 5 patients with the positive RAST test had nasal allergy or signs suggesting allergy. Theree were two middle ear effusions which seemed to be an allergic fluid. However, findings in the present study do not suggest that middle ear effusions result from the direct allergic reaction induced by mites in the mucous membrane lining the middle ear cavity.

Acute Disease↗
PubMed2005

Latex sensitisation and allergy in children with myelomeningocele.

INTRODUCTION: Children with spina bifida (SB) have a high degree of exposure to latex products as a consequence of repeated surgical procedures, implantation of latex-containing materials and catheterisation. The consequence is a higher incidence of latex allergic reactions. OBJECTIVE: The aim of this study is to evaluate the prevalence of latex sensitisation and allergy in a population of children with myelomeningocele (MMC) and to assess the role of associated risk factors. RESULTS: Forty-eight percent of the patients (29 out of 60) showed a latex sensitisation with specific IgE >0.7 kU/l while 15% (9 out of 60) were allergic to latex (specific IgE >0.7 kU/l and clinical manifestations). The principal factor correlated with allergy to latex was specific serum IgE to latex (radioallergosorbent test [RAST]) values (p<0.01). Other factors were total serum IgE (paper radioimmunosorbent test [PRIST]) values, number of surgical procedures and familiarity with allergy. CONCLUSION: These results underline the importance of prophylactic measures to avoid the exposure, not only in the sanitary environment, through the institution of latex-safe routes, but also in daily life, to prevent potentially serious allergic reactions.

Adolescent↗
PubMed1991

Diagnosis of nasal allergy to the house dust mite.

Twenty-five patients with perennial rhinitis and a positive skin prick test (SPT) for Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farinae (Df) were submitted to nasal provocation and the radioimmunosorbent test (RAST) for specific IgE-antibodies. We found a significant correlation in the reaction to both allergen extracts for all parameters examined. In addition, there was a significant correlation among the SPT, the RAST and the nasal provocation for Dp and between the SPT and the RAST for Df. In patients with perennial rhinitis we recommend the combination of all three methods to differentiate unspecific rhinitis from an allergic rhinitis. Only the patients with proved allergic rhinitis could benefit from a specific hyposensitisation.

Adult↗
PubMed1986

Comparison of MAST with radioallergosorbent and skin tests for diagnosis of allergy in children.

The multiple allergosorbent test system (MAST) is a method for measuring total and allergen-specific IgE levels, using autoradiography and densitometry. The purpose of this study was to compare the results of MAST tests with those of radioallergosorbent (RAST) and skin tests as an adjunct to the diagnosis of allergy in children. Twenty children, aged 4 to 19 years, were studied. Total serum IgE level measured by the paper radioimmunosorbent test (PRIST) method ranged from 7 to 1,333 units/mL (geometric mean, 155 units/mL). Total serum IgE level by MAST significantly correlated with the PRIST IgE level. Quantities of allergen-specific IgE measured by MAST and RAST were also significantly correlated. When the diagnostic levels by MAST and RAST were compared with skin test reactions for ragweed, grass, house dust, and mite, MAST had a sensitivity of 59%, a specificity of 97%, and an efficiency of 72%, compared with 67%, 97%, and 78%, respectively, for the RAST analysis. We conclude that MAST and RAST are similar in their ability to measure allergen-specific IgE level, but that neither method is as sensitive as skin tests for detection of allergen-specific IgE.

Adolescent↗
PubMed1999

Breast-feeding, maternal IgE, and total serum IgE in childhood.

BACKGROUND: There is controversy regarding the relationship of the effect of breast-feeding on markers of allergy such as total serum IgE in childhood. OBJECTIVE: This study, using longitudinal data, tested the hypothesis that the relation of breast-feeding to IgE in childhood differs depending on maternal total IgE level. METHODS: Total serum IgE was assessed with the paper radioimmunosorbent test at 4 ages in nonselected children enrolled at birth into the prospective Tucson Children's Respiratory Study. Children were classified as never breast-fed, breast-fed less than 4 months, or breast-fed 4 months or longer, on the basis of physician report or questionnaires completed by parents by the time the child was 18 months old. A longitudinal random effects model was used to test for group differences and temporal trends in IgE for children classified with reference to maternal IgE (high tertile vs all others) and breast-feeding history. A total of 664 children with 1457 observations were included. RESULTS: Among children whose mothers were in the 2 lower tertiles of IgE, breast-feeding was associated with lower total serum IgE at age 6 years (24.2 vs 44.3 IU/mL for never breast-fed children; P <.02); similar trends existed at age 11 years. In contrast, for children whose mothers were in the highest tertile of IgE, breast-feeding of 4 months or longer was associated with higher IgE levels in the child compared with those never breast-fed or breast-fed less than 4 months (97.0 vs 38.9 IU/mL; P <. 005). These cross-sectional analyses were confirmed with the longitudinal random effects model, which also showed no effect of confounders. Paternal IgE showed no similar relation with child IgE. CONCLUSION: Breast-feeding appears to have paradoxic relations with IgE in childhood, depending on maternal IgE level. These findings may help explain the contradictory results found in other investigations of the relation of breast-feeding to allergic symptoms and markers.

Breast Feeding↗
PubMed1975

Measurement of absolute amounts of antigen-specific human IgE by a radioallergosorbent test (RAST) elution technique.

A technique for the absolute quantification of antigen-specific human IgE is described. It employs elution of a calculable amount of antigen-specific IgE from an allergosorbent-antibody complex by means of alkaline pH treatment, followed by measurement of the IgE content of the eluate with a modified radioimmunosorbent test (RIST). With this method IgE antibody directed against the benzylpenicilloyl determinant of penicillin (BPO) was measured quantitatively in sera from seven penicillin allergic patients. IgE specific for ragweed antigen E was measured in sera from 33 ragweed allergic patients. Values obtained for IgE anti-BPO ranged from 19 to 1806 ng/ml and comprised from 1.3 to 27.5% of total serum IgE. Values of IgE anti-antigen E ranged from 9 to 1807 ng/ml, comprising from 3 to 84% of total serum IgE. Excellent correlation (r = 0.99; p less than 0.001) was obtained for both antigen systems between values determined by the RAST elution technique and by simple RAST assay with interpolation from a reference serum of known specific IgE content as determined by the elution technique may be needed only for primary standardization of reference sera.

Allergens↗
PubMed1986

A sensitive and reproducible method for the determination of subnanogram quantities of immunoglobulin E (IgE).

An immunoradiometric assay for immunoglobulin E (IgE) using high adsorption polystyrene test tubes as the solid phase (the 'Maxisorp' assay) is described. The sensitivity of the method was found to be 50 pg IgE/ml and the within-assay reproducibility was 3-7%. The accuracy was estimated by means of a comparison between the Maxisorp assay and paper radioimmunosorbent test (PRIST) and a correlation coefficient of 0.98 (P less than 0.001) was obtained. We conclude that the Maxisorp assay is a fast and reliable assay for IgE determination in cord blood, cell culture supernatants and other highly diluted IgE preparations.

Antibody Specificity↗
PubMed1977

IgE studies on respiratory tract allergies.

The IgE content was determined and specific IgE antibodies were identified in serum samples and nasal secretions of 40 control subjects, 103 patients with nasal allergy, and 37 patients with bronchial asthma using a radioimmunosorbent test an both the serum samples and nasal secretions of allergic patients. Approximately 70% of the serum samples and nasal secretions obtained from patients with positive house dust skin test results were found to contain specific IgE antibodies to mites. The results of this study proved the usefulness of the determination of IgE and specific IgE antibodies in the diagnosis and study of the pathogenesis of respiratory tract allergies. The IgE in nasal secretions was much more closely related to pathogenesis of respiratory allergies than was IgE in serum samples. However, unless more concentrated nasal secretions are assayed, the diagnostic value is limited.

Adolescent↗
PubMed1992

Strong serum inhibition of specific IgE correlated to competing IgG4, revealed by a new methodology in subjects from a S. mansoni endemic area.

A method allowing the immunopurification of human IgE from small volumes of sera with a yield close to 100% (mean = 97.8%; SEM = 0.7) has been developed. The immunopurification eluates were cleared of other class antibodies that could compete with IgE in specific assays. Immunopurification of IgE followed by specific IgE enzyme-linked immunosorbent assay (ELISA) (IMMEL) was then applied to sera of 160 individuals from an area endemic for Schistosoma mansoni. In comparison with radioimmunosorbent test (RAST) and ELISA performed on unfractionated sera, IMMEL provided the highest specific IgE signals. Furthermore, the best correlations between the specific IgE levels and either the specific basophil histamine release levels (r = 0.84; p less than 10(-4) or the anti-S. mansoni skin test values (r = 0.45; p = 10(-4)) were obtained with IMMEL. Measurement of anti-S. mansoni IgE levels in immunopurified fractions and in unfractionated sera of these 160 individuals revealed a strong serum inhibition (geometric means of 98.6% and 96.8% for the adult worms and the larvae, respectively) of the specific IgE reactivity in ELISA. This inhibition was correlated with the anti-adult worm and anti-larval IgG4 levels (r = 0.65; p less than 10(-4) and r = 0.58; p less than 10(-4), respectively). In contrast, this inhibition did not correlate with the specific IgG1, IgG2, IgG3 and IgM levels. Furthermore, the level of specific IgG4 was clearly lower than that of specific IgG1, suggesting that the major contribution of IgG4 in the competition effect is not due to higher levels but rather to a specificity spectrum close to that of the specific IgE. These results support the idea that a specific function of IgG4 in serum might be to control antigen recognition by IgE and consequently, to regulate anaphylactic reactions and IgE-mediated immunity.

Animals↗
PubMed1986

[Immune complexes in allergies: evidence of IgE-containing immune complexes in Churg-Strauss vasculitis].

In five patients with vasculitis, hypereosinophilia and elevated serum-IgE levels, the diagnosis of Churg-Strauss syndrome was established. To identify a possible role of IgE in pathogenic mechanisms leading to vasculitis, a sequential precipitation of patients' sera was performed using various concentrations of polyethylene glycol 6000 (PEG). Using a radioimmunosorbent test, the precipitates obtained were tested for their IgE contents. Considerable amounts of IgE were found in the serum precipitates of all patients. In parallel studies, no IgE-containing precipitates were detected in sera from patients with different allergic diseases and high IgE serum levels. The demonstration of IgE-containing PEG precipitable material in all patients suffering from Churg-Strauss syndrome, together with the finding of an elevated C3d and C4 level and a decreased C3 level in one patient and an increased C3d level in a second patient, suggests that IgE-containing immune complexes play a pathogenic role in the Churg-Strauss syndrome.

Adult↗