Production of a monoclonal antibody specific for ovine immunoglobulin E and its application to monitor serum IgE responses to Haemonchus contortus infection

Parasitology ◽  
1997 ◽  
Vol 114 (4) ◽  
pp. 395-406 ◽  
Author(s):  
F. N. J. KOOYMAN ◽  
P. J. S. VAN KOOTEN ◽  
J. F. HUNTLEY ◽  
A. MacKELLAR ◽  
A. W. C. A. CORNELISSEN ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Haemonchus Contortus ◽  
Immunoglobulin E ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Specific Ige ◽  
Cross Reactivity ◽  
Total Serum ◽  
Western Blots ◽  
Serum Ige

Part of the Cε3–Cε4 region of the ovine immunoglobulin E (IgE) gene (nucleotides 1111–1575) was amplified by PCR. The recombinant protein (recIgE1-2) was expressed in E. coli and both monoclonal and polyclonal antibodies were produced. These antibodies recognized recIgE1-2 and native IgE on Western blots and in ELISA. The polyclonal serum showed cross-reactivity with other sheep immunoglobulin classes. The monoclonal antibody was specific for ovine IgE and goat IgE. Infection of sheep with the abomasal nematode Haemonchus contortus resulted in elevated IgE levels in serum 2–4 weeks after infection, as measured by sandwich ELISA using the rabbit polyclonal as capture antibody and the monoclonal antibody against ovine IgE as second antibody. A negative correlation between worm counts and total serum IgE levels at the end of the experiment was found in repeatedly infected sheep. Significant increased levels of excretory–secretory antigens specific IgE levels were found after H. contortus infection. In contrast, no significant changes in 3rd-stage larvae (L3) antigen-specific IgE titre in sera could be detected after infection.

LINKAGE ANALYSIS OF IL-4 AND OTHER CHROMOSOME 5q31.1 MARKERS AND TOTAL SERUM IMMUNOGLOBULIN E (IgE) CONCENTRATIONS

PEDIATRICS ◽  
1995 ◽  
Vol 96 (2) ◽  
pp. 379-379
Author(s):  
Arden Levy
Keyword(s):  
Linkage Analysis ◽  
Immunoglobulin E ◽  
Specific Ige ◽  
Serum Immunoglobulin ◽  
Total Serum ◽  
Serum Ige ◽  
Sib Pair Analysis ◽  
Pair Analysis

This group found evidence linking five markers in chromosome 5q31.1 with a gene modulating total serum IgE concentration in a sib-pair analysis of 170 individuals in 11 Amish families. Evidence was found for the linkage of 5q31. I and the IL-4 gene. Specific IgE concentrations were not linked to these markers. Thus, this study suggests that IL-4 or a nearby gene in 5q31.1 regulates overall IgE production.

Proficiency Survey-Based Evaluation of Clinical Total and Allergen-Specific IgE Assay Performance

2010 ◽  
Vol 134 (7) ◽  
pp. 975-982 ◽  
Author(s):  
Robert G. Hamilton
Keyword(s):  
Allergic Disease ◽  
Immunoglobulin E ◽  
Diagnostic Algorithm ◽  
Specific Ige ◽  
Total Serum ◽  
Design Data ◽  
Ige Antibody ◽  
Marked Variability ◽  
Specific Immunoglobulin E ◽  
History Of

Abstract Context.—The diagnostic algorithm for human allergic disease involves confirmation of sensitization by detection of allergen-specific immunoglobulin E (IgE) antibody in individuals suspected of having allergic disease because of a history of allergic symptoms after known allergen exposure. Previous studies showed wide disparity among clinically reported allergen-specific IgE levels from different serologic assays. Objective.—To validate the relative analytic performance (sensitivity, interassay reproducibility, linearity/parallelism, intermethod agreement) of clinically used total and allergen-specific IgE assays by using College of American Pathologists' Diagnostic Allergy “SE” Proficiency Survey data. Design.—Data from 2 SE survey cycles were used to assess relative analytic performance of the ImmunoCAP (Phadia), Immulite (Siemens Healthcare-Diagnostics), and HYTEC 288 (HYCOR-Agilent Technologies) total and allergen-specific IgE assays. In each cycle, 2 recalcified plasma pools from atopic donors were diluted twice with IgE-negative serum and evaluated in approximately 200 federally certified clinical laboratories for total IgE and IgE antibody to 5 allergen specificities. Statistical analysis evaluated analytic sensitivity, linearity, reproducibility, and intermethod agreement. Results.—Interlaboratory intramethod, intermethod, and interdilution agreement of all 6 clinically used total serum IgE assays were excellent, with coefficients of variation (CVs) below 15%. Interlaboratory intramethod, and interdilution agreement of 3 clinically used allergen-specific IgE assays were also excellent with CVs below 15%. However, intermethod CVs identified between-assay disagreement greater than 20% in 80% of allergen-specific IgE measurements. Allergen reagents and patients' immune response heterogeneity are suggested probable causes. Conclusions.—Clinical total and allergen-specific IgE assays display excellent analytic sensitivity, precision, reproducibility, and linearity. Marked variability in quantitative estimates of allergen-specific IgE from clinically used automated immunoassays is a concern that may be ameliorated with component allergen use.

scholarly journals Allergenic Characterization of Tropomyosin from the Dusky Brown Cockroach, Periplaneta fuliginosa

2004 ◽  
Vol 11 (4) ◽  
pp. 680-685 ◽  
Author(s):  
Kyoung Yong Jeong ◽  
Heeyu Hwang ◽  
Jongweon Lee ◽  
In-Yong Lee ◽  
Dong Soo Kim ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Periplaneta Americana ◽  
Immunoglobulin E ◽  
Expression System ◽  
Specific Ige ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Inhibitor Concentration ◽  
The Cross ◽  
Elisa Inhibition

ABSTRACTHousehold arthropods are one of the most common causes of allergic diseases. Four species of cockroaches are found to reside in Korean homes, but published work deals almost exclusively with the German and American cockroaches. This study was undertaken to investigate the cross-reactive allergenic components of the dusky brown cockroach,Periplaneta fuliginosa. Enzyme-linked immunosorbent assay (ELISA) inhibition and immunoblot analyses for the dusky brown cockroach were performed withBlattella germanicaandDermatophagoides farinaeallergic sera. cDNA encoding tropomyosin, which is a well known cross-reactive pan-allergen, was cloned by reverse transcriptase PCR, and recombinant protein was produced by using a pET-28b expression system. Native tropomyosin was purified by ammonium sulfate fractionation and electroelution. The immunoglobulin E (IgE) reactivities of native and recombinant tropomyosins were compared by an ELISA inhibition study. All 30 sera tested showedP. fuliginosa-specific IgE, and the IgE-binding reactivity of theP. fuliginosaextract was inhibited as much as 79.4% by aB. germanicaextract and as much as 63.3% by aD. farinaeextract. The deduced amino acid sequence of cloned cDNA was identical with that ofPeriplaneta americanatropomyosin (98.5% nucleotide sequence identity). Seven of 26 (26.9%) allergic sera had IgE specific for recombinant protein, and the maximum inhibition ofP. fuliginosa-specific IgE achieved with recombinant tropomyosin was 37.7% at an inhibitor concentration of 10 μg/ml. Native tropomyosin inhibited the binding of IgE to theP. fuliginosa,B. germanica, andD. farinaeextracts by 65.0, 51.8, and 39% at an inhibitor concentration of 1 μg/ml.P. fuliginosaappears to possess allergens that are highly cross-reactive with allergens ofB. germanicaandD. farinae. Tropomyosin was found to be a major allergenic component accounting for the cross-reactivity between cockroaches and dust mites.

scholarly journals The Study of a Possible Correlation between Serum Levels of Interleukin 17 and Clinical Severity in Patients with Allergic Rhinitis

2017 ◽  
Vol 8 (3) ◽  
pp. ar.2017.8.0207
Author(s):  
Mai Aly Gharib Aly ◽  
Mohamed Tawfik El Tabbakh ◽  
Waheed Fawzy Heissam ◽  
Said Hamed Abbadi
Keyword(s):  
Allergic Rhinitis ◽  
Immunoglobulin E ◽  
Skin Testing ◽  
Allergic Diseases ◽  
Serum Levels ◽  
Clinical Severity ◽  
Total Serum ◽  
Interleukin 17 ◽  
Serum Ige ◽  
Cluster Immunotherapy

Introduction Allergic rhinitis (AR) is one of the most common allergic diseases, which affects ~20% of the world's population. T-helper (Th) type 2 cells produce interleukin (IL) 4 and IL-13, and mediate allergic responses, and these cytokines have been extensively studied as key players in the atopic airway diseases. However, the involvement of Th17 cells and IL-17 in AR has not been clearly examined. Aim To reevaluate AR clinical severity with serum IL-17, whether IL-17 affects the disease alone or in contribution with the atopic predisposition. Patients and Methods During an 18-month period, 39 individuals were divided into three groups: A, (13 control), B (13 with mild-to-moderate AR), and C (13 with severe AR). Both group B and group C patients (26) were subjected to clinical examination and allergy skin testing, and to measurement of both total serum immunoglobulin E (IgE) and IL-17 levels. Eleven patients with AR then were exposed to 6 months of cluster immunotherapy, whereas the rest of the patients were not exposed. Results Revealed a significant elevation of serum IL-17 levels with an associated increase in serum IgE in the patients with AR compared with controls and revealed that the serum levels of both total serum IgE and IL-17 decreased significantly after cluster immunotherapy. Conclusion These preliminary results added new data about the use of injective immunotherapy as well as reported on the use of sublingual immunotherapy.

Characterization of G12 Sandwich ELISA, a Next-Generation Immunoassay for Gluten Toxicity

2012 ◽  
Vol 95 (2) ◽  
pp. 372-376 ◽  
Author(s):  
Elisabeth Halbmayr-Jech ◽  
Elisabeth Hammer ◽  
Richard Fielder ◽  
Jacqueline Coutts ◽  
Adrian Rogers ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Celiac Disease ◽  
Working Group ◽  
Sandwich Elisa ◽  
Extraction Methods ◽  
Cross Reactivity ◽  
Next Generation ◽  
Preliminary Results ◽  
Sample Extraction

Abstract In this work, a monoclonal antibody called G12, raised against the most immunotoxic peptide to celiac disease patients, was used to develop a sandwich ELISA. Preliminary results on cross-reactivities, recoveries, and extraction methods of the new assay are presented. The assay calibration was performed using material from the Prolamin Working Group. The antibody's specificity was determined by cross-reactivity studies on different grains, nuts, oils, and starches. Recovery of the assay was determined by spiking experiments on common food matrixes, as well as on problematic matrixes. Furthermore, sample extraction methods using ethanol, cocktail solution, and a proprietary buffer have been compared.

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