Quantitation of HLA-A*0201 bound tumor associated antigens on a peptide pulsed B cell line

Hum Immunol. 2001 Feb;62(2):125-32. doi: 10.1016/s0198-8859(00)00251-2.

Abstract

CTLs recognize 8- to 10-mer peptides on MHC class I molecules. Recent studies have shown that human CTLs kill autologous tumor cells in an HLA-restricted and peptide-specific manner, and that artificial pep- tides can stimulate tumor-specific CTLs both in vitro and in vivo. Accordingly, several human clinical trials using such peptides are ongoing worldwide. In such methods, the amount of peptide-MHC complexes that remain on the cell surface of APCs after peptide administration is crucial, because CTL activation depends on the number of ligated TCRs and co-stimulation. However, it remains uncertain how many peptide-MHC complexes are reconstituted and remain on live cells after peptide administration. We herein examined the binding affinities of five HLA-A*0201 restricted peptides-four TAAs and one HIV antigen-to HLA-A*0201 molecules and their decay rates on a live B cell line using tandem mass spectrometry. Our experiments showed that nearly 10(5) peptide-MHC complexes per cell could be reconstituted on a cell surface by pulsing a high dose of peptide even if the binding affinities were intermediate or low. However, the decay rates observed for these pep- tide-MHC complexes on a B cell line were faster than previously estimated.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Neoplasm / immunology
  • Antigens, Neoplasm / metabolism*
  • Callithrix
  • Cell Line
  • Cell Membrane / chemistry
  • Cell Membrane / immunology
  • Cell Membrane / metabolism
  • Dose-Response Relationship, Immunologic
  • HIV Reverse Transcriptase / immunology
  • HIV Reverse Transcriptase / metabolism
  • HLA-A2 Antigen / immunology
  • HLA-A2 Antigen / metabolism*
  • Humans
  • Hybridomas
  • Membrane Glycoproteins / immunology
  • Membrane Glycoproteins / metabolism
  • Mice
  • Neoplasm Proteins / immunology
  • Neoplasm Proteins / metabolism
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / immunology
  • Peptide Fragments / metabolism*
  • Protein Binding / immunology
  • Receptor, ErbB-2 / immunology
  • Receptor, ErbB-2 / metabolism
  • Spectrometry, Mass, Electrospray Ionization
  • Tumor Suppressor Protein p53 / immunology
  • Tumor Suppressor Protein p53 / metabolism
  • gp100 Melanoma Antigen

Substances

  • Antigens, Neoplasm
  • HER2-neu-derived peptide (654-662)
  • HLA-A2 Antigen
  • MAGEA3 protein, human
  • Mageb3 protein, mouse
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • PMEL protein, human
  • Peptide Fragments
  • Pmel protein, mouse
  • Tumor Suppressor Protein p53
  • gp100 Melanoma Antigen
  • Receptor, ErbB-2
  • HIV Reverse Transcriptase