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Tissue Antigens. 2012 Apr;79(4):291-4. doi: 10.1111/j.1399-0039.2011.01833.x. Epub 2012 Jan 17.

Allele-specific amplification of the complete HLA-C gene from genomic DNA - a novel Cw4 allele (C*04:71) with a Cw1 motif in the peptide-binding site.

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  • 1Laboratorio de Inmunogenética-HLA, Servicio de Inmunología, Hospital Universitario Puerta de Hierro, Majadahonda, Madrid, Spain.

Abstract

To determine the complete sequence of a newly identified human leukocyte antigen (HLA)-C allele, we designed a method where the full genomic sequence of HLA-C*04 was amplified in isolation from the patient second HLA-C allele in a single polymerase chain reaction (PCR), using primers spanning its 5'- and 3'-untranslated regions. The new allele, officially designated HLA-C*04:71, differs from HLA-C*04:01:01:01 by two single-nucleotide polymorphisms: one determines substitution of phenylalanine for serine 9 at the B pocket of the peptide-binding site; the second substitution is a new polymorphism in intron 5. Phe-9 is characteristic of Cw1 alleles and its presence in C*04:71 most likely affects its peptide-binding repertoire. The principle we have used for C*04:71 isolation could be adapted for unambiguous sequence-based HLA-C typing of selected samples in a clinical setting.

© 2012 John Wiley & Sons A/S.

PMID:
22251067
[PubMed - indexed for MEDLINE]
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