2D TCR-pMHC-CD8 kinetics determines T-cell responses in a self-antigen-specific TCR system

Eur J Immunol. 2014 Jan;44(1):239-50. doi: 10.1002/eji.201343774. Epub 2013 Oct 20.


Two-dimensional (2D) kinetic analysis directly measures molecular interactions at cell-cell junctions, thereby incorporating inherent cellular effects. By comparison, three-dimensional (3D) analysis probes the intrinsic physical chemistry of interacting molecules isolated from the cell. To understand how T-cell tumor reactivity relates to 2D and 3D binding parameters and to directly compare them, we performed kinetic analyses of a panel of human T-cell receptors (TCRs) interacting with a melanoma self-antigen peptide (gp100209 -217 ) bound to peptide-major histocompatibility complex in the absence and presence of co-receptor CD8. We found that while 3D parameters are inadequate to predict T-cell function, 2D parameters (that do not correlate with their 3D counterparts) show a far broader dynamic range and significantly improved correlation with T-cell function. Thus, our data support the general notion that 2D parameters of TCR-peptide-major histocompatibility complex-CD8 interactions determine T-cell responsiveness and suggest a potential 2D-based strategy to screen TCRs for tumor immunotherapy.

Keywords: 2D kinetics; Micropipette adhesion frequency assay; T-cell activation; Thermal fluctuation assay.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD8 Antigens / chemistry
  • CD8 Antigens / metabolism*
  • Cells, Cultured
  • HLA-A2 Antigen / metabolism*
  • Humans
  • Intercellular Junctions / immunology
  • Kinetics
  • Lymphocyte Activation
  • Neoplasms / immunology*
  • Neoplasms / therapy*
  • Protein Binding
  • Receptors, Antigen, T-Cell / chemistry
  • Receptors, Antigen, T-Cell / metabolism*
  • T-Lymphocytes / immunology*
  • gp100 Melanoma Antigen / immunology
  • gp100 Melanoma Antigen / metabolism


  • CD8 Antigens
  • HLA-A2 Antigen
  • PMEL protein, human
  • Receptors, Antigen, T-Cell
  • gp100 Melanoma Antigen