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Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide

Glycosphingolipids containing very-long-chain fatty acids (VLCFAs) regulate several immune responses, such as cytokine production, immune signaling, and antibody induction. We previously reported that stimulation with an inflammatory mediator, TNF-α, promotes the expression of glycosphingolipids in...

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Autor principal: Okuda, Tetsuya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7279471/
https://www.ncbi.nlm.nih.gov/pubmed/32455599
http://dx.doi.org/10.3390/ijms21103632
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author Okuda, Tetsuya
author_facet Okuda, Tetsuya
author_sort Okuda, Tetsuya
collection PubMed
description Glycosphingolipids containing very-long-chain fatty acids (VLCFAs) regulate several immune responses, such as cytokine production, immune signaling, and antibody induction. We previously reported that stimulation with an inflammatory mediator, TNF-α, promotes the expression of glycosphingolipids in vascular endothelial cells. The major component is globotetraosylceramide containing VLCFAs (Gb4Cer-VLCFAs), but its role in inflammatory responses has not been fully investigated. In this study, the antibody-inducing properties of Gb4Cer-VLCFAs were analyzed using serum and hybridoma cells generated from Gb4Cer-VLCFA-immunized mice. The reactivity of serum antibodies against Gb4Cer indicated that immunization with Gb4Cer-VLCFAs immediately induced the production of anti-Gb4Cer antibodies. Over 81% of hybridomas generated from the splenocytes of an immunized mouse produced anti-Gb4Cer antibodies, a subset of which recognized an epitope shared by Gb4Cer and its precursor globotriaosylceramide (Gb3Cer). Further biochemical analyses of established monoclonal antibodies revealed that these antibodies included IgM and IgG3, which specifically react with Gb4Cer and Gb3Cer. These results indicate that immunization with Gb4Cer-VLCFAs can efficiently induce the production of anti-Gb4Cer and -Gb3Cer antibodies by B cells.
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spelling pubmed-72794712020-06-17 Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide Okuda, Tetsuya Int J Mol Sci Communication Glycosphingolipids containing very-long-chain fatty acids (VLCFAs) regulate several immune responses, such as cytokine production, immune signaling, and antibody induction. We previously reported that stimulation with an inflammatory mediator, TNF-α, promotes the expression of glycosphingolipids in vascular endothelial cells. The major component is globotetraosylceramide containing VLCFAs (Gb4Cer-VLCFAs), but its role in inflammatory responses has not been fully investigated. In this study, the antibody-inducing properties of Gb4Cer-VLCFAs were analyzed using serum and hybridoma cells generated from Gb4Cer-VLCFA-immunized mice. The reactivity of serum antibodies against Gb4Cer indicated that immunization with Gb4Cer-VLCFAs immediately induced the production of anti-Gb4Cer antibodies. Over 81% of hybridomas generated from the splenocytes of an immunized mouse produced anti-Gb4Cer antibodies, a subset of which recognized an epitope shared by Gb4Cer and its precursor globotriaosylceramide (Gb3Cer). Further biochemical analyses of established monoclonal antibodies revealed that these antibodies included IgM and IgG3, which specifically react with Gb4Cer and Gb3Cer. These results indicate that immunization with Gb4Cer-VLCFAs can efficiently induce the production of anti-Gb4Cer and -Gb3Cer antibodies by B cells. MDPI 2020-05-21 /pmc/articles/PMC7279471/ /pubmed/32455599 http://dx.doi.org/10.3390/ijms21103632 Text en © 2020 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Okuda, Tetsuya
Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide
title Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide
title_full Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide
title_fullStr Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide
title_full_unstemmed Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide
title_short Isolation and Characterization of Antibodies Induced by Immunization with TNF-α Inducible Globotetraosylceramide
title_sort isolation and characterization of antibodies induced by immunization with tnf-α inducible globotetraosylceramide
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7279471/
https://www.ncbi.nlm.nih.gov/pubmed/32455599
http://dx.doi.org/10.3390/ijms21103632
work_keys_str_mv AT okudatetsuya isolationandcharacterizationofantibodiesinducedbyimmunizationwithtnfainducibleglobotetraosylceramide