Dataset Information

Identification of IgE sequential epitopes of lentil (Len c 1) by means of peptide microarray immunoassay.

ABSTRACT:

Background

Lentils are often responsible for allergic reactions to legumes in Mediterranean children. Although the primary sequence of the major allergen Len c 1 is known, the location of the IgE-binding epitopes remains undefined.

Objective

We sought to identify IgE-binding epitopes of Len c 1 and relate epitope binding to clinical characteristics.

Methods

One hundred thirty-five peptides corresponding to the primary sequence of Len c 1 were probed with sera from 33 patients with lentil allergy and 15 nonatopic control subjects by means of microarray immunoassay. Lentil-specific IgE levels, skin prick test responses, and clinical reactions to lentil were determined. Epitopes were defined as overlapping signal above interslide and intraslide cutoffs and confirmed by using inhibition assays with a peptide from the respective region. Hierarchic clustering of microarray data was used to correlate binding patterns with clinical findings.

Results

The patients with lentil allergy specifically recognized IgE-binding epitopes located in the C-terminal region between peptides 107 and 135. Inhibition experiments confirmed the specificity of IgE binding in this region, identifying different epitopes. Linkage of cluster results with clinical data and lentil-specific IgE levels displayed a positive correlation between lentil-specific IgE levels, epitope recognition, and respiratory symptoms. Modeling based on the 3-dimensional structure of a homologous soy vicilin suggests that the Len c 1 epitopes identified are exposed on the surface of the molecule.

Conclusion

Several IgE-binding sequential epitopes of Len c 1 have been identified. Epitopes are located in the C-terminal region and are predicted to be exposed on the surface of the protein. Epitope diversity is positively correlated with IgE levels, pointing to a more polyclonal IgE response.

SUBMITTER: Vereda A

PROVIDER: S-EPMC2948849 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Publications

Identification of IgE sequential epitopes of lentil (Len c 1) by means of peptide microarray immunoassay.

Vereda Andrea A Andreae Doerthe A DA Lin Jing J Shreffler Wayne G WG Ibañez Maria Dolores MD Cuesta-Herranz Javier J Bardina Ludmilla L Sampson Hugh A HA

The Journal of allergy and clinical immunology 20100901 3

<h4>Background</h4>Lentils are often responsible for allergic reactions to legumes in Mediterranean children. Although the primary sequence of the major allergen Len c 1 is known, the location of the IgE-binding epitopes remains undefined.<h4>Objective</h4>We sought to identify IgE-binding epitopes of Len c 1 and relate epitope binding to clinical characteristics.<h4>Methods</h4>One hundred thirty-five peptides corresponding to the primary sequence of Len c 1 were probed with sera from 33 patien ...[more]

PMID: 20816193

Similar Datasets

Project description:IntroductionImmunoinformatic tools can be used to predict schistosome-specific B-cell epitopes with little sequence identity to human proteins and antigens other than the target. This study reports an approach for identifying schistosome peptides mimicking linear B-cell epitopes using in-silico tools and peptide microarray immunoassay validation.MethodFirstly, a comprehensive literature search was conducted to obtain published schistosome-specific peptides and recombinant proteins with the best overall diagnostic performances. For novel peptides, linear B-cell epitopes were predicted from target recombinant proteins using ABCpred, Bcepred and BepiPred 2.0 in-silico tools. Together with the published peptides, predicted peptides with the highest probability of being B-cell epitopes and the lowest sequence identity with proteins from human and other pathogens were selected. Antibodies against the peptides were measured in sera, using peptide microarray immunoassays. Area under the ROC curve was calculated to assess the overall diagnostic performances of the peptides.ResultsPeptide AA81008-19-30 had excellent and acceptable diagnostic performances for discriminating S. mansoni and S. haematobium positives from healthy controls, with AUC values of 0.8043 and 0.7326 respectively for IgG. Peptides MS3_10186-123-131, MS3_10385-339-354, SmSPI-177-193, SmSPI-379-388, MS3-10186-40-49 and SmS-197-214 had acceptable diagnostic performances for discriminating S. mansoni positives from healthy controls with AUC values ranging from 0.7098 to 0.7763 for IgG. Peptides SmSPI-359-372, Smp126160-438-452 and MS3 10186-25-41 had acceptable diagnostic performances for discriminating S. mansoni positives from S. mansoni negatives with AUC values of 0.7124, 0.7156 and 0.7115 respectively for IgG. Peptide MS3-10186-40-49 had an acceptable diagnostic performance for discriminating S. mansoni positives from healthy controls, with an AUC value of 0.7413 for IgM.ConclusionOne peptide with a good diagnostic performance and nine peptides with acceptable diagnostic performances were identified using the immunoinformatic approach and peptide microarray validation. There is need for evaluation of the peptides with true negatives and a good standard positive reference.

Dataset Information

Identification of IgE sequential epitopes of lentil (Len c 1) by means of peptide microarray immunoassay.

Background

Objective

Methods

Results

Conclusion

Publications

Identification of IgE sequential epitopes of lentil (Len c 1) by means of peptide microarray immunoassay.

Similar Datasets

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