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- Title
IgE, IgG1, and IgG4 Reactivity to Dermatophagoides pteronyssinus Glycosylated Extract in Allergic Patients.
- Authors
Resende, Rafael de Oliveira; Ynoue, Leandro Hideki; Miranda, Juliana Silva; de Almeida, Karine Cristine; Silva, Deise Aparecida de Oliveira; Sopelete, Monica Camargo; Alves, Ronaldo; Gennari-Cardoso, Margareth Leitão; Taketomi, Ernesto Akio
- Abstract
Background. House dust mites are important allergen sources and some of these allergenic proteins may contain carbohydrate moieties, which are able to be isolated using lectins, as Concanavalin A (ConA). This study aimed to investigate allergenicity (IgE) and antigenicity (IgG1 and IgG4) of ConA-unbound and ConA-bound Dermatophagoides pteronyssinus (Dpt) crude extracts using sera of mite-allergic patients as well as inhibition capacity of antibody binding. Material and Methods. We obtained mannose-enriched and mannose-depleted fractions from Dpt by ConA affinity chromatography. Both ConA-bound and ConA-unbound fractions were evaluated by ELISA and Western Blotting for specific IgE, IgG1, and IgG4 reactivity with sera obtained from 95 mite-allergic patients (DP+) and 92 nonallergic (NA) subjects. Inhibition ELISA was used to assess cross-reactivity between Dpt extract and its fractions. Results. Among the DP+ patients, no difference was found between ConA-unbound and ConA-bound fractions regarding the levels of specific IgE, IgG1, and IgG4. Nonallergic subjects had the same levels of specific IgG1 to both ConA-unbound and ConA-bound fractions, although for specific IgG4, values were higher for ConA-bound. A positive correlation was found among specific IgE, IgG1, and IgG4 levels when Dpt was compared to ConA-unbound and ConA-bound fractions. Recognition of crude Dpt by IgE, IgG1, and IgG4 was highly inhibited by ConA-unbound and ConA-bound fractions. Western Blotting revealed a broad spectrum of bands ranging from 14 to 116 kDa recognized by specific IgE and IgG4. However, IgG1 reached higher frequency values on high molecular weight polypeptides. Conclusion. ConA-unbound and ConA-bound fractions derived from D. pteronyssinus crude extract revealed important components involved in the IgE recognition in allergic patients as well as IgG1 and/or IgG4 in allergic and healthy subjects.
- Subjects
PLANT protein analysis; ALLERGENS; ALLERGIES; ANIMAL experimentation; ENZYME-linked immunosorbent assay; GLYCOSYLATION; IMMUNOGLOBULINS; LIQUID chromatography; MITES; PEPTIDES; PLANT proteins; TISSUE extracts; WESTERN immunoblotting; HEXOSES
- Publication
BioMed Research International, 2019, p1
- ISSN
2314-6133
- Publication type
Article
- DOI
10.1155/2019/9840890