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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Allergy 49 (1994), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The prevalence and specificity of naturally occurring human IgA anti-IgE autoantibodies (a-E Ab) were studied by ELISA with anti-IgA monoclonal antibodies (mAb) and a purified myeloma IgE as solid-phase protein, i.e., IgE-DES(κ). Such detected IgA a-E Ab were common among adults, and significantly increased geometric means (GM) were found in patients with atopy (P= 0.006; n= 41; GM = 79.3 arbitrary units (AU)/ml) and filariasis (P= 0.02; n= 41; GM = 75.9 AU/ml), as compared with nonatopic controls (n= 42; GM = 48.8 AU/ml). No such difference was observed between age-matched nonatopic (n= 22; GM = 36.7 AU/ml) and atopic (n= 22; GM = 38.6 AU/ml) children. Children had significantly (P= 0.001) lower IgA a-E Ab concentrations than adults, probably as a result of age, because IgA a-E Ab concentrations and age of children were significantly correlated (n= 44; P〈0.05; rs= 0.30). IgA a-E Ab concentrations were very low in cord serum (n= 32; median 〈0.1 AU/ml). Sex did not influence IgA a-E Ab concentrations in any study group. The specificity of IgA a-E Ab in nine sera was studied by ELISA inhibition assay using IgE-DES myeloma as solid-phase protein and inhibitory proteins of the IgG, IgM, IgD, and IgE classes, including five different IgE myeloma proteins, as well as three enzymatic fragments of IgE-DES. The inhibitions indicated that all IgA a-E Ab tested reacted in a low-affinity reaction with determinants restricted to IgE-DES, i.e., the solid-phase protein. These epitopes were heat-resistant (2 h; 56°C) and located in the Fabɛ-DES fragment. No isotype-specific IgA a-E Ab were found because none of the four other IgE proteins were inhibitory. Subclass typing indicated that most IgA a-E Ab belonged to the IgA 1 subclass. It is unlikely, for reasons of restricted specificity, low affinity, and common prevalence, that such IgA 1 a-E Ab are connected with IgE-mediated disorders. The study also raises questions on the definition of anti-IgE antibodies.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Allergy 47 (1992), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Comparison was made of IgE and IgA levels in aspirated and gravity-collected cord blood (CB) from the umbilical vein and in capillary blood samples collected on the 4-5th day of life among 21 infants with atopic heredity. The IgA levels, but not the IgE levels, were significantly (p〈 0.001) lower at days 4-5 of life than at delivery. Further, there were significantly (p〈0.05) more infants with decreasing IgA levels (20/21; 95%) than IgE levels (9/15; 60% of those with detectable IgE, i.e., ≥ 0.125 kU/1). These observations, together with the highly significant correlation observed between IgE in aspirated CB samples and at 4–5 days of age, suggest active IgE synthesis during the prenatal and postnatal periods. Contamination of CB with maternal blood, defined as an increased CBIgA level (≥ 14.1 μg/ml), was found in 3 (14%) CB samples, all of which were gravity-collected. Of 4 CB samples (gravity-collected) with elevated IgE (i.e., ≥ 0.9kU/l), 2 had suspected maternal contamination. Therefore, aspiration of CB or capillary collection at 4-5 days of age should be preferred for allergy prediction. If gravity collection is used, contamination should be investigated by determining IgA in all CB samples with IgE concentrations exceeding the cut-off point.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: A case of monoclonal IgE type lambda with IgE levels about 1 mg/ml has been followed for 6 years. Except for a slight asthma no signs of malignancies, parasitic infestations or other known diseases compatible with hyper-IgE have been found. By the combination of fractional ammonium sulphate precipitation, gel filtration, chromatofocusing, and subtraction affinity chromatography the IgE protein was isolated in an immunochemically pure and homogeneous form. Immunofluorescence of bone marrow cells showed about 1 % IgE plasma cells. The amount of basophil bound IgE was 42 ng/106 cells, and histamine release from basophils challenged with anti-IgE was not different from that in atopic control persons, indicating a within-allergic-patients normal amount of IgE receptors. The protein-A reactivity was 0.4% equivalent to well-known IgE myeloma proteins. No antigen specificity of the IgE protein was found. Only a few cases of asymptomatic hyper-IgE are known, and it cannot be ruled out that this represents a premyeloma condition, since a similar case terminated in a malignant lymphoma.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, U.K. and Cambridge, USA : Blackwell Science Ltd
    Scandinavian journal of immunology 44 (1996), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: In this paper, the authors report the use of liquid-liquid partition chromatography (LLPC) in an aqueous polyethylene glycol (PEG)/dextran two-phase system to compare the surface properties (partition properties) of human antibodies and fragments thereof. The surface properties of all the monoclonal antibodies of different classes and subclasses investigated were within the same broad range as that observed for the polyclonal antibodies and no relationship was found between the exposed surfaces of the immunoglobulins (Ig) and their heavy chain isotype. Moreover, Fc fragments from various IgG1, 2 and 4 myeloma proteins were found to exhibit similar surface properties. Employing chimeric antibodies with identical variable regions the authors found that intact IgG1, 2 and 4 displayed identical surface properties, while the corresponding IgA1, IgA2, IgG3, IgE and IgM antibodies differed both from each other and from the IgGs. The surface properties of chimeric IgG3 could be made similar to those of the IgG1, 2 and 4 chimers by partially reducing the length of the hinge section, but new differences in surface properties appeared when their hinges were of similar length. Thus, LLPC can be used to detect differences or similarities in the surface properties of the antigen-binding regions as well as the Fc part in the various isotypes. This can shed light on biological activities such as antigen binding and effector function.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 11 (1981), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Total IgE was determined in 107 sera using a novel, automated, non-isotopic immunoassay called PACIA (particle-counting immunoassay) based on agglutination of anti-IgE coated latex particles by IgE. The IgE values ranged from 10 to 50 000 iu/ml and were compared with results obtained by a conventional radioimmunoassay (RIA) which used a fast double antibody separation technique the coefficient of correlation was 0.985 and the regression line r = 0-82 x+ 130.00.PACIA had several advantages over the RIA technique: using a sampling rate of 50.hr. results were obtained in 35 min compared to 16-20 hr. no labelled IgE was required and the separation step, which relied on measuring the number of non-agglutinated particles by an optical cell counter, was fully automated. The threshold of sensitivity was 10 iu/ml and the maximal coefficient of variation for between assay precision was 12-9%.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 20 (1990), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The levels of IgG1, IgG2, IgG3 and IgG4 were analysed by FLISA in cord serum and in serum samples collected at 6 and 18 months of age from infants whose mothers were atopic. None of the four IgG subclasses was significantly influenced on any sampling occasion by infant atopy, gender, month of birth, maternal IgE or maternal diet during pregnancy and early lactation. However, at 18 months of age, significantly higher levels of IgG1 (P 〈 0·05) and of IgG4 (P 〈 0·01) were found in infants with an elevated IgE (〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:09547894:CEA407:ges" location="ges.gif"/〉 8·0 kU/1) than in those with a lower level. A weak positive correlation (rs= 0·26; P= 0·05) between IgE and IgG4 was also observed. Despite the fact that the serum levels of IgG4 at 18 months were significantly higher (P 〈 0·01) among infants with positive IgE-RAST (〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:09547894:CEA407:ges" location="ges.gif"/〉 0·15 PRU/ml) to ovomucoid or β-lactoglobulin, our data suggest that the the concentration of IgG4 relates more to the level of IgE than to the clinical symptoms of atopy. Determination of IgG subclasses seems to be of limited value for predicting atopy during early infancy.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Allergy 44 (1989), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The relation between platelet counts (PCT) and IgE was studied in cord blood from 136 European newborns. PCT was significantly lower (P= 0.0014) when cord-IgE was superior to 1.20 IU/ml (n= 29; 245,000/μl) than when it was inferior to this value (n= 107; 284,080/μl) which resulted in a significant negative Spearman rank correlation between PCT and cord-IgE (P= 0.002; rs=−0.25). A follow-up by questionnaire in 97 of the newborns revealed that those newborns who had developed definite atopy within 18 months of age had significantly (P= 0.002) lower PCT at birth (n= 8; 196,000/μl) than those free of atopic symptoms (n= 61; 286,000/μl). Further newborns to atopic mothers (n= 23; 245,000/μl) had significantly (P= 0.014) lower PCT than newborns to non-atopic mothers (n= 74; 286,000/μl). The lowest PCT was recorded when both the mother was atopic and the newborn had developed definite or probable atopy by the age of 18 months (n= 7; 175,000/μl) as compared to atopy alone in mothers (n= 16; 276,000/μl; P= 0.005), to atopy alone in infants (n= 9; 281,000/μ1; P= 0.005) and to non-atopic infants of non-atopic mothers (n= 65; 286,000/μl; P= 0.0007). Significantly (P= 0.03) lower PCT amongst boys (n= 49; 259,000/μl) compared with girls (n= 48; 294,000/μl) was attributed to the higher incidence of elevated cord-IgE and infant atopy among boys. These findings were limited to PCT, since the erythrocyte count, hematocrit and hemoglobin concentration in cord blood did not significantly (P 〉 0.10) differ in any of these groups. Paternal atopy and atopy in siblings did not influence PCT, neither did maternal smoking or drug intake (progesterone and β-mimetics) during pregnancy. These data as a whole are compatible with a direct or indirect role for platelets and their mediators in immediate hypersensitivity reactions. We speculate that a low PCT in cord blood of newborns reflects an on-going intra-uterine sensitization and therefore it might be a complementary parameter to family history and cord-IgE in predicting newborns at high risk of developing atopy.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: We report the development of a three-layer immunoradiometric assay (TIRA) for measurement of IgG antibodies of all four subclasses in human sera. The first layer consists of diluted human serum, the second layer is monoclonal mouse antibodies to human IgG subclasses., and the third layer is 125I-labelled rabbit anti-mouse IgG. Monoclonal anti-IgG1. anti-IgC3 and anti-IgG4 reacted only with their complementary IgG subclass, whereas the anti-lgG2 showed slight cross-reactivity to immunoglobins of other subclasses and classes and to light chain proteins. The observed cross-reactivity was found to be without importance, when the TIRA was applied to measurement of IgG subclass antibodies. Equipotency was established by use of appropriate dilutions of the monoclonal antibodies, and the assay was calibrated by use of human reference serum. The TIRA therefore permits reliable inter-individual and intra-individual comparisons of the IgG antibody response in all four subclasses. Von-sped fie binding obtained with pooled normal human serum was below 0.33′#. Inter-assay coefficient of variation was between 18 and 27%, The TIRA was applied to measurement of IgG subclass antibodies to timothy grass pollen in sera from grass pollen allergies undergoing immunotherapy.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: specificity of IgE binding to a human basophil-like cell line (KU812) was studied by flow cytometry. Four IgE myeloma proteins, representing both light-chain types, one chimeric IgE protein, and polyclonal serum IgE blocked the direct binding of FITC-labeled IgE(DES) myeloma protein to KU812 cells in a dose-dependent and nearly equimolar way. Although not as efficiently as human IgE (from five to eight times less on a molar basis), both rat and mouse IgE blocked IgE(DES)-FITC binding to KU812 cells. In sharp contrast, all four human IgG subclasses, both IgA subclasses, and IgM myeloma proteins, as well as monomeric and heat-aggregated polyclonal human IgG, were unable to block significantly IgE(DES)-FITC binding to KU812 cells (≤0.5/0 on a molar basis). The cytophilic epitope on IgE was heat-susceptible (56 °h), lost after reduction alkylation, and resident in the papain-derived Fcɛ -fragment, but not in the papain-derived F(ab') 2ɛ.- and Fcɛ-fragments nor in the pepsin-derived F(ab')2ɛ- and Fcɛ.'-fragments. Washing and displacement experiments indicated that a major part of IgE reacted with high affinity to KU812 cells. The results indicate that the binding of IgE to KU812 cells is highly specific and involves the classical high-affinity FcɛRI-receptor. Although the density of receptors is low, this human cell line offers a unique model to study IgE/FcɛRI interactions.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: AN automated particle counting immunoassay (PACIA) for measurement of blocking antibodies (antigen neutralizing capacity) against timothy grass pollen extract in sera from desensitized allergies is described.Latex particles coated with E(ab')2-anti-timothy are agglutinated by timothy. Serum containing anti-timothy antibodies inhibits the agglutination. Non-agglutinated particles are counted in a modified Auto Counter. Nineteen of 20 sera from timothy allergies who had undergone immunotherapy with purified timothy extract for 30 weeks, showed significant agglutination-inhibition. None of 42 normal human sera gave significant inhibition. The inhibiting antibody could be removed by absorption with protein A and was thus of non- IgE nature, i.e. blocking antibody. The results obtained correlated statistically significantly with those found with a double-antibody method (rS= 0.62, n= 20, t= 3.35, P 〈 0.01) and with the cumulated dosage of timothy allergen extract administered to the individual patient (rS= 0.56, n= 20, t= 2.87, P 〈0.02). Between-assay coefficient of variation was from 6.4% to 18.3%. The capacity is 40 samples per hour. The method has also been applied to measurement of blocking antibodies to boney bee and wasp venom.
    Type of Medium: Electronic Resource
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