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Defining KIR and HLA Class I Genotypes at Highest Resolution via High-Throughput Sequencing.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Norman, Paul J 
Hollenbach, Jill A 
Nemat-Gorgani, Neda 
Marin, Wesley M 
Norberg, Steven J 

Abstract

The physiological functions of natural killer (NK) cells in human immunity and reproduction depend upon diverse interactions between killer cell immunoglobulin-like receptors (KIRs) and their HLA class I ligands: HLA-A, HLA-B, and HLA-C. The genomic regions containing the KIR and HLA class I genes are unlinked, structurally complex, and highly polymorphic. They are also strongly associated with a wide spectrum of diseases, including infections, autoimmune disorders, cancers, and pregnancy disorders, as well as the efficacy of transplantation and other immunotherapies. To facilitate study of these extraordinary genes, we developed a method that captures, sequences, and analyzes the 13 KIR genes and HLA-A, HLA-B, and HLA-C from genomic DNA. We also devised a bioinformatics pipeline that attributes sequencing reads to specific KIR genes, determines copy number by read depth, and calls high-resolution genotypes for each KIR gene. We validated this method by using DNA from well-characterized cell lines, comparing it to established methods of HLA and KIR genotyping, and determining KIR genotypes from 1000 Genomes sequence data. This identified 116 previously uncharacterized KIR alleles, which were all demonstrated to be authentic by sequencing from source DNA via standard methods. Analysis of just two KIR genes showed that 22% of the 1000 Genomes individuals have a previously uncharacterized allele or a structural variant. The method we describe is suited to the large-scale analyses that are needed for characterizing human populations and defining the precise HLA and KIR factors associated with disease. The methods are applicable to other highly polymorphic genes.

Description

Keywords

HLA class I, KIR, gene content diversity, highly polymorphic, immunity, Alleles, Gene Dosage, Genes, MHC Class I, Genome, Human, Genotype, HLA-A Antigens, HLA-B Antigens, HLA-C Antigens, Haplotypes, High-Throughput Nucleotide Sequencing, Humans, Polymorphism, Genetic, Receptors, KIR

Journal Title

Am J Hum Genet

Conference Name

Journal ISSN

0002-9297
1537-6605

Volume Title

Publisher

Elsevier BV
Sponsorship
This study was supported by U.S. National Institutes of Health grants U01 AI090905, R01 20 GM109030, R01 AI17892 and U19 AI119350. Authors Steven Norberg and Mostafa Ronaghi are 21 employees of Illumina Inc.