Responses of peptide-specific T cells to stimulation with polystyrene beads carrying HLA class I molecules loaded with single peptides

Alberto Chersi, Rossella Galati, Daniele Accapezzato, Vittorio Francavilla, Vincenzo Barnaba, Richard H. Butler, Nobuyuki Tanigaki

Research output: Contribution to journalArticlepeer-review

Abstract

Cell-sized microbeads carrying single peptide-loaded HLA class I molecules were prepared for HLA-A2 and HLA-B7 by a simple procedure which transfers single peptide-loaded HLA class I molecules from cultured cells to polystyrene beads using anti-peptide antibodies directed to an intracellular segment of HLA-A α chains. The surface density of peptide-loaded HLA class I molecules on beads was comparable to that on the peptide-loaded cells. HLA-A2 beads loaded with an HCV peptide HCV1073 were tested for stimulation activity on an HCV1073-specific CD8+ T cell clone NS3-1. A substantial level of γ-IFN production was induced. The stimulation was peptide-specific. The efficiency was dependent on the bead concentration and the surface HLA class I density on beads and enhanced significantly by co-coupling of anti-CD28 to peptide-loaded beads. The peptide-loading efficiency on HLA class I molecules and the transfer efficiency of HLA class I molecules to polystyrene beads were reasonably high for HLA-A2 and HLA-B7. Thus, polystyrene beads carrying these single peptide-loaded HLA class I molecules are potentially useful in further analysis of the co-stimulatory or inhibitory factors involved in CD8+ T cell responses and eventually in detection of cytotoxic T cells in PBLs.

Original languageEnglish
Pages (from-to)79-91
Number of pages13
JournalJournal of Immunological Methods
Volume291
Issue number1-2
DOIs
Publication statusPublished - Aug 2004

Keywords

  • Anti-HLA class I peptide antibody
  • HLA class I microbeads
  • T cell stimulation

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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