Novel multiplex PCR-SSP method for centromeric KIR allele discrimination

Abstract Allelic diversity of the KIR2DL receptors drive differential expression and ligand-binding affinities that impact natural killer cell function and patient outcomes for diverse cancers. We have developed a global intermediate resolution amplification-refractory mutation system (ARMS) PCR-SSP...

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Autores principales:	Jean-Benoît Le Luduec, Anupa Kudva, Jeanette E. Boudreau, Katharine C. Hsu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Killer Cell Immunoglobulin-like Receptors KIR3DL1 Alleles Receptor KIR2DL4 Amplification-refractory Mutation System (ARMS) Centre d’Etude Polymorphisme Humain (CEPH) Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ec063dbd8c6741ea9219a3cc99a4b074
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spelling	oai:doaj.org-article:ec063dbd8c6741ea9219a3cc99a4b0742021-12-02T11:41:03ZNovel multiplex PCR-SSP method for centromeric KIR allele discrimination10.1038/s41598-018-33135-12045-2322https://doaj.org/article/ec063dbd8c6741ea9219a3cc99a4b0742018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-33135-1https://doaj.org/toc/2045-2322Abstract Allelic diversity of the KIR2DL receptors drive differential expression and ligand-binding affinities that impact natural killer cell function and patient outcomes for diverse cancers. We have developed a global intermediate resolution amplification-refractory mutation system (ARMS) PCR-SSP method for distinguishing functionally relevant subgroups of the KIR2DL receptors, as defined by phylogenetic study of the protein sequences. Use of the ARMS design makes the method reliable and usable as a kit, with all reactions utilizing the same conditions. Six reactions define six subgroups of KIR2DL1; four reactions define three subgroups of KIR2DL2; and five reactions define four subgroups of KIR2DL3. Using KIR allele data from a cohort of 426 European-Americans, we identified the most common KIR2DL subtypes and developed the high-throughput PCR-based methodology, which was validated on a separate cohort of 260 healthy donors. Linkage disequilibrium analysis between the different KIR2DL alleles revealed that seven allelic combinations represent more than 95% of the observed population genotypes for KIR2DL1/L2/L3. In summary, our findings enable rapid typing of the most common KIR2DL receptor subtypes, allowing more accurate prediction of co-inheritance and providing a useful tool for the discrimination of observed differences in surface expression and effector function among NK cells exhibiting disparate KIR2DL allotypes.Jean-Benoît Le LuduecAnupa KudvaJeanette E. BoudreauKatharine C. HsuNature PortfolioarticleKiller Cell Immunoglobulin-like ReceptorsKIR3DL1 AllelesReceptor KIR2DL4Amplification-refractory Mutation System (ARMS)Centre d’Etude Polymorphisme Humain (CEPH)MedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-9 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Killer Cell Immunoglobulin-like Receptors KIR3DL1 Alleles Receptor KIR2DL4 Amplification-refractory Mutation System (ARMS) Centre d’Etude Polymorphisme Humain (CEPH) Medicine R Science Q
spellingShingle	Killer Cell Immunoglobulin-like Receptors KIR3DL1 Alleles Receptor KIR2DL4 Amplification-refractory Mutation System (ARMS) Centre d’Etude Polymorphisme Humain (CEPH) Medicine R Science Q Jean-Benoît Le Luduec Anupa Kudva Jeanette E. Boudreau Katharine C. Hsu Novel multiplex PCR-SSP method for centromeric KIR allele discrimination
description	Abstract Allelic diversity of the KIR2DL receptors drive differential expression and ligand-binding affinities that impact natural killer cell function and patient outcomes for diverse cancers. We have developed a global intermediate resolution amplification-refractory mutation system (ARMS) PCR-SSP method for distinguishing functionally relevant subgroups of the KIR2DL receptors, as defined by phylogenetic study of the protein sequences. Use of the ARMS design makes the method reliable and usable as a kit, with all reactions utilizing the same conditions. Six reactions define six subgroups of KIR2DL1; four reactions define three subgroups of KIR2DL2; and five reactions define four subgroups of KIR2DL3. Using KIR allele data from a cohort of 426 European-Americans, we identified the most common KIR2DL subtypes and developed the high-throughput PCR-based methodology, which was validated on a separate cohort of 260 healthy donors. Linkage disequilibrium analysis between the different KIR2DL alleles revealed that seven allelic combinations represent more than 95% of the observed population genotypes for KIR2DL1/L2/L3. In summary, our findings enable rapid typing of the most common KIR2DL receptor subtypes, allowing more accurate prediction of co-inheritance and providing a useful tool for the discrimination of observed differences in surface expression and effector function among NK cells exhibiting disparate KIR2DL allotypes.
format	article
author	Jean-Benoît Le Luduec Anupa Kudva Jeanette E. Boudreau Katharine C. Hsu
author_facet	Jean-Benoît Le Luduec Anupa Kudva Jeanette E. Boudreau Katharine C. Hsu
author_sort	Jean-Benoît Le Luduec
title	Novel multiplex PCR-SSP method for centromeric KIR allele discrimination
title_short	Novel multiplex PCR-SSP method for centromeric KIR allele discrimination
title_full	Novel multiplex PCR-SSP method for centromeric KIR allele discrimination
title_fullStr	Novel multiplex PCR-SSP method for centromeric KIR allele discrimination
title_full_unstemmed	Novel multiplex PCR-SSP method for centromeric KIR allele discrimination
title_sort	novel multiplex pcr-ssp method for centromeric kir allele discrimination
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/ec063dbd8c6741ea9219a3cc99a4b074
work_keys_str_mv	AT jeanbenoitleluduec novelmultiplexpcrsspmethodforcentromerickirallelediscrimination AT anupakudva novelmultiplexpcrsspmethodforcentromerickirallelediscrimination AT jeanetteeboudreau novelmultiplexpcrsspmethodforcentromerickirallelediscrimination AT katharinechsu novelmultiplexpcrsspmethodforcentromerickirallelediscrimination
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Novel multiplex PCR-SSP method for centromeric KIR allele discrimination

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