Differential presentation of tumor antigen-derived epitopes by MHC-class I and antigen-positive tumor cells

Int J Cancer. 2008 Oct 15;123(8):1841-7. doi: 10.1002/ijc.23728.

Abstract

SSX2 is a member of the family of cancer/testis antigens. The SSX2 derived peptide SSX2(103-111) has been shown to be presented to cytotoxic T-lymphocytes (CTL) by Major-Histocompatibility (MHC) Class-I complexes after endogenous processing, more precisely by the allele HLA-A*0201. The HLA-A*0201- and SSX2-positive melanoma cell line SK-Mel-37 but not Me275 had been shown to elicit reactivity in SSX2(103-111) specific cytotoxic T-lymphocytes. To analyze the correlation between SSX2(103-111) presentation and T-cell stimulation, we intended to visualize presentation of SSX2(103-111) in these melanoma cell lines. Fab-antibodies were established from a human phage library with specificity for SSX2(103-111)/HLA-A*0201 complexes (but non-reactive with HLA-A*0201 or SSX2(103-111) alone) and used to visualize the presentation of SSX2(103-111) in the context of HLA-A*0201 by fluorescence microscopy. Presentation of SSX2(103-111) the context of HLA-A*0201 was demonstrated for the majority of SK-Mel-37, but for only a small fraction (<1%) of Me275 as indicated by a clear membrane-staining pattern in fluorescence microscopy. The presentation of SSX2(103-111) on SK-Mel37 and Me275, but not the expression of the SSX2 protein correlated with the capability of these cells to stimulate cells of an SSX2(103-111)-specific T-cell clone. MHC-peptide specific antibodies are a valuable tool for the analysis of antigenic peptides in the context of MHC-I molecules and for the structural definition of immunodominant epitopes.

MeSH terms

  • 3T3 Cells
  • Animals
  • Antibody Specificity
  • Antigen Presentation
  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / immunology*
  • Breast Neoplasms / genetics
  • Breast Neoplasms / immunology*
  • Cell Line, Tumor
  • HLA-A Antigens / immunology*
  • HLA-A2 Antigen
  • Humans
  • Immunoglobulin Fragments / immunology
  • Melanoma / genetics
  • Melanoma / immunology*
  • Mice
  • Microscopy, Fluorescence
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / immunology*
  • Peptide Fragments / immunology
  • Polymerase Chain Reaction
  • Repressor Proteins / biosynthesis
  • Repressor Proteins / genetics
  • Repressor Proteins / immunology*
  • T-Lymphocytes, Cytotoxic / immunology

Substances

  • Antigens, Neoplasm
  • HLA-A Antigens
  • HLA-A*02:01 antigen
  • HLA-A2 Antigen
  • Immunoglobulin Fragments
  • Neoplasm Proteins
  • Peptide Fragments
  • Repressor Proteins
  • synovial sarcoma X breakpoint proteins