Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study

One of the purposes of specific immunotherapy (SIT) is to modulate humoral immune response against allergens with significant increases in allergen-specific IgG levels, commonly associated with blocking activity. The present study investigated in vitro blocking activity of allergen-specific IgG anti...

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Main Authors: Isabella Lima Siman, Lais Martins de Aquino, Leandro Hideki Ynoue, Juliana Silva Miranda, Ana Claudia Arantes Marquez Pajuaba, Jair Pereira Cunha-Júnior, Deise Aparecida Oliveira Silva, Ernesto Akio Taketomi
Format: Article
Language:English
Published: Wiley 2013-01-01
Series:Clinical and Developmental Immunology
Online Access:http://dx.doi.org/10.1155/2013/657424
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author Isabella Lima Siman
Lais Martins de Aquino
Leandro Hideki Ynoue
Juliana Silva Miranda
Ana Claudia Arantes Marquez Pajuaba
Jair Pereira Cunha-Júnior
Deise Aparecida Oliveira Silva
Ernesto Akio Taketomi
author_facet Isabella Lima Siman
Lais Martins de Aquino
Leandro Hideki Ynoue
Juliana Silva Miranda
Ana Claudia Arantes Marquez Pajuaba
Jair Pereira Cunha-Júnior
Deise Aparecida Oliveira Silva
Ernesto Akio Taketomi
author_sort Isabella Lima Siman
collection DOAJ
description One of the purposes of specific immunotherapy (SIT) is to modulate humoral immune response against allergens with significant increases in allergen-specific IgG levels, commonly associated with blocking activity. The present study investigated in vitro blocking activity of allergen-specific IgG antibodies on IgE reactivity to Dermatophagoides pteronyssinus (Dpt) in sera from atopic patients. Dpt-specific IgG antibodies were purified by ammonium sulfate precipitation followed by protein-G affinity chromatography. Purity was checked by SDS-PAGE and immunoreactivity by slot-blot and immunoblot assays. The blocking activity was evaluated by inhibition ELISA. The electrophoretic profile of the ammonium sulfate precipitated fraction showed strongly stained bands in ligand fraction after chromatography, compatible with molecular weight of human whole IgG molecule. The purity degree was confirmed by detecting strong immunoreactivity to IgG, negligible to IgA, and no reactivity to IgE and IgM. Dpt-specific IgG fraction was capable of significantly reducing levels of IgE anti-Dpt, resulting in 35%–51% inhibition of IgE reactivity to Dpt in atopic patients sera. This study showed that allergen-specific IgG antibodies purified from mite-allergic patients sera block the IgE recognition of Dermatophagoides pteronyssinus antigens. This approach reinforces that intermittent measurement of serum allergen-specific IgG antibodies will be an important objective laboratorial parameter that will help specialists to follow their patients under SIT.
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spelling doaj-art-d2bc69c9889b4c3b8cd947cd550a1add2025-08-20T03:24:07ZengWileyClinical and Developmental Immunology1740-25221740-25302013-01-01201310.1155/2013/657424657424Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro StudyIsabella Lima Siman0Lais Martins de Aquino1Leandro Hideki Ynoue2Juliana Silva Miranda3Ana Claudia Arantes Marquez Pajuaba4Jair Pereira Cunha-Júnior5Deise Aparecida Oliveira Silva6Ernesto Akio Taketomi7Laboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilLaboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilLaboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilLaboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilLaboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilLaboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilLaboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilLaboratory of Allergy and Clinical Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia, Avenida Pará 1720, Bloco 4 C, Campus Umuarama, 38400-902 Uberlândia, MG, BrazilOne of the purposes of specific immunotherapy (SIT) is to modulate humoral immune response against allergens with significant increases in allergen-specific IgG levels, commonly associated with blocking activity. The present study investigated in vitro blocking activity of allergen-specific IgG antibodies on IgE reactivity to Dermatophagoides pteronyssinus (Dpt) in sera from atopic patients. Dpt-specific IgG antibodies were purified by ammonium sulfate precipitation followed by protein-G affinity chromatography. Purity was checked by SDS-PAGE and immunoreactivity by slot-blot and immunoblot assays. The blocking activity was evaluated by inhibition ELISA. The electrophoretic profile of the ammonium sulfate precipitated fraction showed strongly stained bands in ligand fraction after chromatography, compatible with molecular weight of human whole IgG molecule. The purity degree was confirmed by detecting strong immunoreactivity to IgG, negligible to IgA, and no reactivity to IgE and IgM. Dpt-specific IgG fraction was capable of significantly reducing levels of IgE anti-Dpt, resulting in 35%–51% inhibition of IgE reactivity to Dpt in atopic patients sera. This study showed that allergen-specific IgG antibodies purified from mite-allergic patients sera block the IgE recognition of Dermatophagoides pteronyssinus antigens. This approach reinforces that intermittent measurement of serum allergen-specific IgG antibodies will be an important objective laboratorial parameter that will help specialists to follow their patients under SIT.http://dx.doi.org/10.1155/2013/657424
spellingShingle Isabella Lima Siman
Lais Martins de Aquino
Leandro Hideki Ynoue
Juliana Silva Miranda
Ana Claudia Arantes Marquez Pajuaba
Jair Pereira Cunha-Júnior
Deise Aparecida Oliveira Silva
Ernesto Akio Taketomi
Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study
Clinical and Developmental Immunology
title Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study
title_full Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study
title_fullStr Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study
title_full_unstemmed Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study
title_short Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study
title_sort allergen specific igg antibodies purified from mite allergic patients sera block the ige recognition of dermatophagoides pteronyssinus antigens an in vitro study
url http://dx.doi.org/10.1155/2013/657424
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