Flow-cytometric determination of peptide-class I complex formation identification of p53 peptides that bind to HLA-A2

Herbert J. Zeh, Gerhard H. Leder, Michael T. Lotze, Russell D. Salter, Matt Tector, György Stuber, Susanne Modrow, Walter J. Storkus

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

A novel class I-peptide-binding assay was developed and used to identify a series of peptides derived from the human p53 tumor-suppressor gene product capable of binding the HLA-A2 class I allele. Brief pH 3.3 acid treatment of human cell lines rapidly denatures pre-existing class I complexes, as detected by loss of binding of conformation-dependent mAbs, leaving only free class I heavy chains associated with the viable cell surface. These heavy chains may be induced to refold and be recognized by antibodies (in 2-4 hours) when acid-treated cells are coincubated with exogenous ß2-microglobulin and peptides capable of binding the relevant class I allele examined. This assay, with a detection limit of 1-10 nM peptide, was used to screen the capacity of a panel of nine peptides bearing HLA-A2-binding motifs and derived from the human p53 tumor-suppressor protein sequence. Eight of the nine peptides bound to, and reconstituted, HLA-A2 on acid-treated cells. This assay system will enable the rapid identification of peptides binding to any class I allele, which is the initial prerequisite for elucidating potential CD8 + T-cell epitopes.

Original languageEnglish (US)
Pages (from-to)79-86
Number of pages8
JournalHuman Immunology
Volume39
Issue number2
DOIs
StatePublished - Feb 1994
Externally publishedYes

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HLA-A2 Antigen
Peptides
Alleles
Acids
Tumor Suppressor Protein p53
T-Lymphocyte Epitopes
Tumor Suppressor Genes
Limit of Detection
Cell Line
Antibodies

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Flow-cytometric determination of peptide-class I complex formation identification of p53 peptides that bind to HLA-A2. / Zeh, Herbert J.; Leder, Gerhard H.; Lotze, Michael T.; Salter, Russell D.; Tector, Matt; Stuber, György; Modrow, Susanne; Storkus, Walter J.

In: Human Immunology, Vol. 39, No. 2, 02.1994, p. 79-86.

Research output: Contribution to journalArticle

Zeh, Herbert J. ; Leder, Gerhard H. ; Lotze, Michael T. ; Salter, Russell D. ; Tector, Matt ; Stuber, György ; Modrow, Susanne ; Storkus, Walter J. / Flow-cytometric determination of peptide-class I complex formation identification of p53 peptides that bind to HLA-A2. In: Human Immunology. 1994 ; Vol. 39, No. 2. pp. 79-86.
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