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Authors: Creary, Lisa E and Guerra, Sandra G and Chong, Winnie and Brown, Colin J and Turner, Thomas R and Robinson, James and Bultitude, Will P and Mayor, Neema P and Marsh, Steven G E and Saito, Katsuyuki and Lam, Kevin and Duke, Jamie L and Mosbruger, Timothy L and Ferriola, Deborah and Monos, Dimitrios and Willis, Amanda and Askar, Medhat and Fischer, Gottfried and Saw, Chee Loong and Ragoussis, Jiannis and Petrek, Martin and Serra-Pagés, Carles and Juan, Manel and Stavropoulos-Giokas, Catherine and Dinou, Amalia and Ameen, Reem and Al Shemmari, Salem and Spierings, Eric and Gendzekhadze, Ketevan and Morris, Gerald P and Zhang, Qiuheng and Kashi, Zahra and Hsu, Susan and Gangavarapu, Sridevi and Mallempati, Kalyan C and Yamamoto, Fumiko and Osoegawa, Kazutoyo and Vayntrub, Tamara and Chang, Chia-Jung and Hansen, John A and Fernández-Vina, Marcelo A

Extended molecular characterization of HLA genes in the IHWG reference B-lymphoblastoid cell lines (B-LCLs) was one of the major goals for the 17th International HLA and Immunogenetics Workshop (IHIW). Although reference B-LCLs have been examined extensively in previous workshops complete high-resolution typing was not completed for all the classical class I and class II HLA genes. To address this, we conducted a single-blind study where select panels of B-LCL genomic DNA samples were distributed to multiple laboratories for HLA genotyping by next-generation sequencing methods. Identical cell panels comprised of 24 and 346 samples were distributed and typed by at least four laboratories in order to derive accurate consensus HLA genotypes. Overall concordance rates calculated at both 2- and 4-field allele-level resolutions ranged from 90.4% to 100%. Concordance for the class I genes ranged from 91.7 to 100%, whereas concordance for class II genes was variable; the lowest observed at HLA-DRB3 (84.2%). At the maximum allele-resolution 78 B-LCLs were defined as homozygous for all 11 loci. We identified 11 novel exon polymorphisms in the entire cell panel. A comparison of the B-LCLs NGS HLA genotypes with the HLA genotypes catalogued in the IPD-IMGT/HLA Database Cell Repository, revealed an overall allele match at 68.4%. Typing discrepancies between the two datasets were mostly due to the lower-resolution historical typing methods resulting in incomplete HLA genotypes for some samples listed in the IPD-IMGT/HLA Database Cell Repository. Our approach of multiple-laboratory NGS HLA typing of the B-LCLs has provided accurate genotyping data. The data generated by the tremendous collaborative efforts of the 17th IHIW participants is useful for updating the current cell and sequence databases and will be a valuable resource for future studies.Copyright © 2019. Published by Elsevier Inc.

Journal: Human immunology
DOI: 10.1016/j.humimm.2019.03.001
Year: 2019

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