Characterization of + cell subsets with a monoclonal antibody selectively recognizing and blocking the specific interaction with
Meazza, Raffaella (IRCCS Ospedale Policlinico San Martino (Gènova, Itàlia))
Falco, Michela (IRCCS Istituto Giannina Gaslini (Gènova, Itàlia))
Canevali, Paolo (IRCCS Ospedale Policlinico San Martino (Gènova, Itàlia))
Loiacono, Fabrizio (IRCCS Ospedale Policlinico San Martino (Gènova, Itàlia))
Colomar-Carando, Natalia (University of Genoa)
Muntasell i Castellví, Aura 1972- (Institut Hospital del Mar d'Investigacions Mèdiques)
Rea, Anna (Universitat Pompeu Fabra)
Mingari, Maria Cristina (University of Genoa)
Locatelli, Franco (Università degli Studi di Roma "La Sapienza")
Moretta, Lorenzo (Ospedale Pediatrico Bambino Gesù (Roma, Itàlia))
López-Botet, Miguel (Universitat Pompeu Fabra)
Pende, Daniela (IRCCS Ospedale Policlinico San Martino (Gènova, Itàlia))
Universitat Autònoma de Barcelona

Data:	2022
Resum:	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.
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	HLA-C ; Killer immunoglobulin-like receptors ; KIR/KIR-ligand interaction ; Monoclonal antibodies ; Natural killer cells
Publicat a:	Hla, Vol. 100 (may 2022) , p. 119-132, ISSN 2059-2310

DOI: 10.1111/tan.14640
PMID: 35439359

14 p, 2.8 MB

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