Novel cellular microarray assay for profiling T-cell peptide antigen specificities

C. Yue, M. Oelke, M. E. Paulaitis, J. P. Schneck

Research output: Contribution to journalArticlepeer-review

Abstract

We present a novel cellular microarray assay using soluble peptide-loaded HLA A2-Ig dimer complexes that optimizes the avidity of peptide-HLA binding by preserving the molecular flexibility of the dimer complex while attaining much higher concentrations of the complex relative to cognate T-cell receptors. A seminal advance in assay development is made by separating the molecular T-cell receptor recognition event from the binding interactions that lead to antigen-specific cell capture on the microarray. This advance enables the quantitative determination of antigen-specific frequencies in heterogeneous T-cell populations without enumerating the number of cells captured on the microarray. The specificity of cell capture, sensitivity to low antigen-specific frequencies, and quantitation of antigenic T-cell specificities are established using CD8 T-cell populations with prepared antigen-specific CTL frequencies and heterogeneous T cells isolated from peripheral blood. The results demonstrate several advantages for high-throughput broad-based, quantitative assessments of low-frequency antigen specificities. The assay enables the use of cellular microarrays to determine the stability and flux of antigen-specific T-cell responses within and across populations.

Original languageEnglish (US)
Pages (from-to)5629-5637
Number of pages9
JournalJournal of proteome research
Volume9
Issue number11
DOIs
StatePublished - Nov 5 2010

Keywords

  • HLA-Ig
  • T cells
  • cellular microarray
  • high-throughput screening
  • human leukocyte antigen
  • immune response
  • infectious diseases
  • influenza
  • vaccines

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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