Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes

Killer immunoglobulin-like receptor (KIR) genes code for a family of inhibitory and activating receptors, finely tuning NK cell function. Numerous studies reported the relevance of KIR allelic polymorphism on KIR expression, ligand affinity, and strength in signal transduction. Although KIR variabil...

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Autores: Falco, Michela, Meazza, Raffaella, Alicata, Claudia, Canevali, Paolo, Muntasell i Castellví, Aura, 1972-, Bottino, Cristina, Moretta, Lorenzo, Pende, Daniela, López-Botet, M. (Miguel)
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2022
País:España
Recursos:Universitat Pompeu Fabra
Repositório:Repositorio Digital de la UPF
OAI Identifier:oai:repositori.upf.edu:10230/54129
Acesso em linha:http://hdl.handle.net/10230/54129
http://dx.doi.org/10.1111/tan.14630
Access Level:Acceso aberto
Palavra-chave:Killer immunoglobulin-like receptors
Monoclonal antibodies
Natural killer cells
Polymorphism
Site-directed mutagenesis
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spelling Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypesFalco, MichelaMeazza, RaffaellaAlicata, ClaudiaCanevali, PaoloMuntasell i Castellví, Aura, 1972-Bottino, CristinaMoretta, LorenzoPende, DanielaLópez-Botet, M. (Miguel)Killer immunoglobulin-like receptorsMonoclonal antibodiesNatural killer cellsPolymorphismSite-directed mutagenesisKiller immunoglobulin-like receptor (KIR) genes code for a family of inhibitory and activating receptors, finely tuning NK cell function. Numerous studies reported the relevance of KIR allelic polymorphism on KIR expression, ligand affinity, and strength in signal transduction. Although KIR variability, including gene copy number and allelic polymorphism, in combination with HLA class I polymorphism, impacts both KIR expression and NK cell education, only a precise phenotypic analysis can define the size of the different KIRpos NK cell subsets. In this context, reagents recognizing a limited number of KIRs is essential. In this study, we have characterized the specificity of an anti-KIR mAb termed HP-DM1. Testing its binding to HEK-293T cells transfected with plasmids coding for different KIRs, we demonstrated that HP-DM1 mAb exclusively reacts with KIR2DL1. Using site-directed mutagenesis, we identified the four amino acids relevant for HP-DM1 recognition: M44, S67, R68, and T70. HP-DM1 mAb binds to a conformational epitope including M44, the residue crucial for HLA-C K80 recognition by KIR2DL1. Based on the HP-DM1 epitope characterization, we could extend its reactivity to all KIR2DL1 allotypes identified except for KIR2DL1*022 and, most likely, KIR2DL1*020, predicting that it does not recognize any other KIR with the only exception of KIR2DS1*013. Moreover, by identifying the residues relevant for HP-DM1 binding, continuously updating of its reactivity will be facilitated.This work was supported by Italian Ministry of Health (Ricerca Corrente G. Gaslini, Ricerca Corrente Ospedale Policlinico San Martino) (Cristina Bottino and Daniela Pende), Instituto de Salud Carlos III, FIS00/0181 (to Miguel Lopez-Botet), H2020-MSCA-ITN-2017-765104-MATURE-NK (to Miguel Lopez-Botet and Daniela Pende), AIRC 5×1000 2018 id. 21147 (Lorenzo Moretta).Wiley202220222022info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/54129http://dx.doi.org/10.1111/tan.14630reponame:Repositorio Digital de la UPFinstname:Universitat Pompeu FabraInglésHLA. 2022 Aug;100(2):107-18info:eu-repo/grantAgreement/EC/H2020/765104© 2022 The Authors. HLA: Immune Response Genetics published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:repositori.upf.edu:10230/541292026-06-12T07:21:37Z
dc.title.none.fl_str_mv Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
title Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
spellingShingle Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
Falco, Michela
Killer immunoglobulin-like receptors
Monoclonal antibodies
Natural killer cells
Polymorphism
Site-directed mutagenesis
title_short Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
title_full Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
title_fullStr Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
title_full_unstemmed Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
title_sort Epitope characterization of a monoclonal antibody that selectively recognizes KIR2DL1 allotypes
dc.creator.none.fl_str_mv Falco, Michela
Meazza, Raffaella
Alicata, Claudia
Canevali, Paolo
Muntasell i Castellví, Aura, 1972-
Bottino, Cristina
Moretta, Lorenzo
Pende, Daniela
López-Botet, M. (Miguel)
author Falco, Michela
author_facet Falco, Michela
Meazza, Raffaella
Alicata, Claudia
Canevali, Paolo
Muntasell i Castellví, Aura, 1972-
Bottino, Cristina
Moretta, Lorenzo
Pende, Daniela
López-Botet, M. (Miguel)
author_role author
author2 Meazza, Raffaella
Alicata, Claudia
Canevali, Paolo
Muntasell i Castellví, Aura, 1972-
Bottino, Cristina
Moretta, Lorenzo
Pende, Daniela
López-Botet, M. (Miguel)
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Killer immunoglobulin-like receptors
Monoclonal antibodies
Natural killer cells
Polymorphism
Site-directed mutagenesis
topic Killer immunoglobulin-like receptors
Monoclonal antibodies
Natural killer cells
Polymorphism
Site-directed mutagenesis
description Killer immunoglobulin-like receptor (KIR) genes code for a family of inhibitory and activating receptors, finely tuning NK cell function. Numerous studies reported the relevance of KIR allelic polymorphism on KIR expression, ligand affinity, and strength in signal transduction. Although KIR variability, including gene copy number and allelic polymorphism, in combination with HLA class I polymorphism, impacts both KIR expression and NK cell education, only a precise phenotypic analysis can define the size of the different KIRpos NK cell subsets. In this context, reagents recognizing a limited number of KIRs is essential. In this study, we have characterized the specificity of an anti-KIR mAb termed HP-DM1. Testing its binding to HEK-293T cells transfected with plasmids coding for different KIRs, we demonstrated that HP-DM1 mAb exclusively reacts with KIR2DL1. Using site-directed mutagenesis, we identified the four amino acids relevant for HP-DM1 recognition: M44, S67, R68, and T70. HP-DM1 mAb binds to a conformational epitope including M44, the residue crucial for HLA-C K80 recognition by KIR2DL1. Based on the HP-DM1 epitope characterization, we could extend its reactivity to all KIR2DL1 allotypes identified except for KIR2DL1*022 and, most likely, KIR2DL1*020, predicting that it does not recognize any other KIR with the only exception of KIR2DS1*013. Moreover, by identifying the residues relevant for HP-DM1 binding, continuously updating of its reactivity will be facilitated.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/54129
http://dx.doi.org/10.1111/tan.14630
url http://hdl.handle.net/10230/54129
http://dx.doi.org/10.1111/tan.14630
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv HLA. 2022 Aug;100(2):107-18
info:eu-repo/grantAgreement/EC/H2020/765104
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:Repositorio Digital de la UPF
instname:Universitat Pompeu Fabra
instname_str Universitat Pompeu Fabra
reponame_str Repositorio Digital de la UPF
collection Repositorio Digital de la UPF
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.812429

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