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Characterization of KIR + NK cell subsets with a monoclonal antibody selectively recognizing KIR2DL1 and blocking the specific interaction with HLA-C

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dc.contributor.author Meazza, Raffaella
dc.contributor.author Falco, Michela
dc.contributor.author Canevali, Paolo
dc.contributor.author Loiacono, Fabrizio
dc.contributor.author Colomar-Carando, Natalia
dc.contributor.author Muntasell i Castellví, Aura, 1972-
dc.contributor.author Rea, Anna
dc.contributor.author Mingari, Maria Cristina
dc.contributor.author Locatelli, Franco
dc.contributor.author Moretta, Lorenzo
dc.contributor.author López-Botet, Miguel
dc.contributor.author Pende, Daniela
dc.date.accessioned 2022-09-21T06:13:06Z
dc.date.available 2022-09-21T06:13:06Z
dc.date.issued 2022
dc.identifier.citation Meazza R, Falco M, Canevali P, Loiacono F, Colomar-Carando N, Muntasell A, Rea A, Mingari MC, Locatelli F, Moretta L, Lopez-Botet M, Pende D. Characterization of KIR + NK cell subsets with a monoclonal antibody selectively recognizing KIR2DL1 and blocking the specific interaction with HLA-C. HLA. 2022 Aug;100(2):119-32. DOI: 10.1111/tan.14640
dc.identifier.issn 2059-2302
dc.identifier.uri http://hdl.handle.net/10230/54128
dc.description.abstract The phenotypic identification of different NK cell subsets allows more in-depth characterization of KIR repertoire and function, which are of potential interest in KIR and disease association studies. KIR genes are highly polymorphic, but a great homology exists among the various sequences and few monoclonal antibodies (mAbs) specifically recognize a single KIR. This is the case of HP-DM1 which was demonstrated by analysis of cell transfectants and epitope mapping to be exclusively KIR2DL1-specific, covering all allotypes identified to date, except for KIR2DL1*022 and *020, and also to react with KIR2DS1*013. Here, we compared in immunofluorescence analyses the staining of HP-DM1 with other available mAbs to precisely identify KIR2DL1+ NK cells in potential donors for αβT/B-depleted haplo-HSCT, with known KIR genotype. HP-DM1 mAb was used in combination with EB6 or 11PB6 (anti-KIR2DL1/S1 and anti-KIR2DL3*005), 143211 (anti-KIR2DL1/S5), and HP-MA4 (anti-KIR2DL1/S1/S3/S5) mAbs, allowing the accurate identification of different KIR+ NK cell subsets. These phenotypic evaluations appeared useful to dissect the expression pattern of various KIR2D in NK cells from KIR2DL3*005+ individuals, particularly if KIR2DS1 is present. HP-DM1 mAb remarkably refined NK cell phenotyping of donors carrying KIR2DS5, either in the centromeric or telomeric region. Functional assays with KIR2DL1+ /S1+ /S5+ NK cells confirmed that only HP-DM1 exclusively reacts with KIR2DL1. Finally, we demonstrated that HP-DM1 mAb blocked KIR2DL1 recognition of C2+ HLA-C. Altogether, the data support that HP-DM1 is a unique reagent valuable for characterizing KIR+ NK cell subsets.
dc.description.sponsorship This work was supported by a network grant of the European Commission (H2020-MSCA-ITN-765104-MATURE-NK) to Miguel Lopez-Botet and Daniela Pende; Natalia Colomar-Carando and Anna Rea are fellows in the project. This work was also supported by Italian Ministry of Health (Ricerca Corrente G. Gaslini, Ricerca Corrente and 5xmille-2016 Ospedale Policlinico San Martino), Fondazione AIRC 5 per mille 2018 id 21147 to Franco Locatelli and Lorenzo Moretta.
dc.format.mimetype application/pdf
dc.language.iso eng
dc.publisher Wiley
dc.relation.ispartof HLA. 2022 Aug;100(2):119-32
dc.rights © 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 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
dc.rights.uri http://creativecommons.org/licenses/by/4.0/
dc.title Characterization of KIR + NK cell subsets with a monoclonal antibody selectively recognizing KIR2DL1 and blocking the specific interaction with HLA-C
dc.type info:eu-repo/semantics/article
dc.identifier.doi http://dx.doi.org/10.1111/tan.14640
dc.subject.keyword HLA-C
dc.subject.keyword KIR/KIR-ligand interaction
dc.subject.keyword Killer immunoglobulin-like receptors
dc.subject.keyword Monoclonal antibodies
dc.subject.keyword Natural killer cells
dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/765104
dc.rights.accessRights info:eu-repo/semantics/openAccess
dc.type.version info:eu-repo/semantics/publishedVersion

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