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J Immunol Methods. 2014 Jan 31;403(1-2):72-8. doi: 10.1016/j.jim.2013.11.023. Epub 2013 Dec 3.

Combinatorial HLA-peptide bead libraries for high throughput identification of CD8⁺ T cell specificity.

Author information

1
MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.

Abstract

Comprehensive antigenic characterization of a T cell population of unknown specificity is challenging. Existing MHC class I expression systems are limited by the practical difficulty of probing cell populations with an MHC class I peptide library and the cross-reactivity of T cells that are able to recognise many variants of an index peptide. Using emulsion PCR and emulsion in vitro transcription/translation of a random library of peptides conjugated to CD8-null HLA-A*0201 on beads, we probed HLA-A*0201-restricted T cells with specificity for influenza, CMV and EBV. We observed significant enrichment for sequences containing HLA-A2 anchors and correct viral fragments for all T cell populations. HLA bead display provides a novel approach to identify the specificity of T cells.

KEYWORDS:

Bead display; HLA library; T cells

PMID:
24309405
DOI:
10.1016/j.jim.2013.11.023
[Indexed for MEDLINE]

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