Defining the cross-reactivity between peanut allergens Ara h 2 and Ara h 6 using monoclonal antibodies
© The Author(s) 2024. Published by Oxford University Press on behalf of the British Society for Immunology. All rights reserved. For permissions, please e-mail: journals.permissionsoup.com. Elements of the work have been written by employees of the US Government..
In peanut allergy, Arachis hypogaea 2 (Ara h 2) and Arachis hypogaea 6 (Ara h 6) are two clinically relevant peanut allergens with known structural and sequence homology and demonstrated cross-reactivity. We have previously utilized X-ray crystallography and epitope binning to define the epitopes on Ara h 2. We aimed to quantitatively characterize the cross-reactivity between Ara h 2 and Ara h 6 on a molecular level using human monoclonal antibodies (mAbs) and structural characterization of allergenic epitopes. We utilized mAbs cloned from Ara h 2 positive single B cells isolated from peanut-allergic, oral immunotherapy-treated patients to quantitatively analyze cross-reactivity between recombinant Ara h 2 (rAra h 2) and Ara h 6 (rAra h 6) proteins using biolayer interferometry and indirect inhibitory ELISA. Molecular dynamics simulations assessed time-dependent motions and interactions in the antibody-antigen complexes. Three epitopes-conformational epitopes 1.1 and 3, and the sequential epitope KRELRNL/KRELMNL-are conserved between Ara h 2 and Ara h 6, while two more conformational and three sequential epitopes are not. Overall, mAb affinity was significantly lower to rAra h 6 than it was to rAra h 2. This difference in affinity was primarily due to increased dissociation of the antibodies from rAra h 6, a phenomenon explained by the higher conformational flexibility of the Ara h 6-antibody complexes in comparison to Ara h 2-antibody complexes. Our results further elucidate the cross-reactivity of peanut 2S albumins on a molecular level and support the clinical immunodominance of Ara h 2.
Medienart: |
E-Artikel |
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Erscheinungsjahr: |
2024 |
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Erschienen: |
2024 |
Enthalten in: |
Zur Gesamtaufnahme - volume:216 |
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Enthalten in: |
Clinical and experimental immunology - 216(2024), 1 vom: 12. März, Seite 25-35 |
Sprache: |
Englisch |
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Beteiligte Personen: |
Marini-Rapoport, Orlee [VerfasserIn] |
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Links: |
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Anmerkungen: |
Date Completed 13.03.2024 Date Revised 09.04.2024 published: Print Citation Status MEDLINE |
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doi: |
10.1093/cei/uxae005 |
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funding: |
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Förderinstitution / Projekttitel: |
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PPN (Katalog-ID): |
NLM368362531 |
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520 | |a In peanut allergy, Arachis hypogaea 2 (Ara h 2) and Arachis hypogaea 6 (Ara h 6) are two clinically relevant peanut allergens with known structural and sequence homology and demonstrated cross-reactivity. We have previously utilized X-ray crystallography and epitope binning to define the epitopes on Ara h 2. We aimed to quantitatively characterize the cross-reactivity between Ara h 2 and Ara h 6 on a molecular level using human monoclonal antibodies (mAbs) and structural characterization of allergenic epitopes. We utilized mAbs cloned from Ara h 2 positive single B cells isolated from peanut-allergic, oral immunotherapy-treated patients to quantitatively analyze cross-reactivity between recombinant Ara h 2 (rAra h 2) and Ara h 6 (rAra h 6) proteins using biolayer interferometry and indirect inhibitory ELISA. Molecular dynamics simulations assessed time-dependent motions and interactions in the antibody-antigen complexes. Three epitopes-conformational epitopes 1.1 and 3, and the sequential epitope KRELRNL/KRELMNL-are conserved between Ara h 2 and Ara h 6, while two more conformational and three sequential epitopes are not. Overall, mAb affinity was significantly lower to rAra h 6 than it was to rAra h 2. This difference in affinity was primarily due to increased dissociation of the antibodies from rAra h 6, a phenomenon explained by the higher conformational flexibility of the Ara h 6-antibody complexes in comparison to Ara h 2-antibody complexes. Our results further elucidate the cross-reactivity of peanut 2S albumins on a molecular level and support the clinical immunodominance of Ara h 2 | ||
650 | 4 | |a Journal Article | |
650 | 4 | |a Research Support, N.I.H., Intramural | |
650 | 4 | |a Research Support, N.I.H., Extramural | |
650 | 4 | |a Ara h 2 | |
650 | 4 | |a Ara h 6 | |
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700 | 1 | |a Fernández-Quintero, Monica L |e verfasserin |4 aut | |
700 | 1 | |a Keswani, Tarun |e verfasserin |4 aut | |
700 | 1 | |a Zong, Guangning |e verfasserin |4 aut | |
700 | 1 | |a Shim, Jane |e verfasserin |4 aut | |
700 | 1 | |a Pedersen, Lars C |e verfasserin |4 aut | |
700 | 1 | |a Mueller, Geoffrey A |e verfasserin |4 aut | |
700 | 1 | |a Patil, Sarita U |e verfasserin |4 aut | |
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