IgE, IgG1, and IgG4 Reactivity to Dermatophagoides pteronyssinus Glycosylated Extract in Allergic Patients

• Published in: BioMed research international Vol. 2019; no. 2019; pp. 1 - 12
• Main Authors: Taketomi, Ernesto Akio, Gennari-Cardoso, Margareth Leitão, Sopelete, Monica Camargo, Silva, Deise Aparecida Oliveira, de Almeida, Karine Cristine, Miranda, Juliana Silva, Ynoue, Leandro Hideki, Resende, Rafael de Oliveira, Alves, Ronaldo
• Format: Journal Article
• Language: English
• Published: Cairo, Egypt Hindawi Publishing Corporation 2019; Hindawi; John Wiley & Sons, Inc
• Subjects: Adult; Affinity chromatography; Allergenicity; Allergens; Allergens - chemistry; Allergens - immunology; Allergies; Animals; Antibodies; Antigenicity; Antigens; Archives & records; Carbohydrates; Chromatography; Cloning; Concanavalin A; Concanavalin A - chemistry; Concanavalin A - immunology; Cross-reactivity; Dermatophagoides pteronyssinus; Dermatophagoides pteronyssinus - chemistry; Dermatophagoides pteronyssinus - immunology; Enzyme-Linked Immunosorbent Assay; Female; Glycosylation; Healthy Volunteers; House dust; Humans; Hypersensitivity - immunology; Immunoglobulin E; Immunoglobulin E - chemistry; Immunoglobulin E - immunology; Immunoglobulin G; Immunoglobulin G - chemistry; Immunoglobulin G - immunology; Immunoglobulins; Immunology; Inhibition; Lectins; Male; Mannose; Medical diagnosis; Middle Aged; Mites; Molecular weight; Plant Extracts - chemistry; Plant Extracts - immunology; Polypeptides; Proteins; Pyroglyphidae - chemistry; Pyroglyphidae - immunology; Reactivity; Recognition; Western blotting; Brazil; United States > US
• ISSN: 2314-6133; 2314-6141; 2314-6141
• DOI: 10.1155/2019/9840890

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.