Clustering of class I HLA oligomers with CD8 and TCR: three-dimensional models based on fluorescence resonance energy transfer and crystallographic data

J Immunol. 2001 Apr 15;166(8):5078-86. doi: 10.4049/jimmunol.166.8.5078.

Abstract

Fluorescence resonance energy transfer (FRET) data, in accordance with lateral mobility measurements, suggested the existence of class I HLA dimers and oligomers at the surface of live human cells, including the B lymphoblast cell line (JY) used in the present study. Intra- and intermolecular class I HLA epitope distances were measured on JY B cells by FRET using fluorophore-conjugated Ag-binding fragments of mAbs W6/32 and L368 directed against structurally well-characterized heavy and light chain epitopes, respectively. Out-of-plane location of these epitopes relative to the membrane-bound BODIPY-PC (2-(4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine) was also determined by FRET. Computer-simulated docking of crystallographic structures of class I HLA and epitope-specific Ag-binding fragments, with experimentally determined interepitope and epitope to cell surface distances as constraints, revealed several sterically allowed and FRET-compatible class I HLA dimeric and tetrameric arrangements. Extension of the tetrameric class I HLA model with interacting TCR and CD8 resulted in a model of a supramolecular cluster that may exist physiologically and serve as a functionally significant unit for a network of CD8-HLA-I complexes providing enhanced signaling efficiency even at low MHC-peptide concentrations at the interface of effector and APCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen-Presenting Cells / chemistry
  • Antigen-Presenting Cells / immunology
  • B-Lymphocytes / chemistry
  • B-Lymphocytes / immunology
  • CD8 Antigens / chemistry*
  • Cell Line, Transformed
  • Cell Membrane / chemistry
  • Cell Membrane / immunology
  • Computer Simulation
  • Crystallography, X-Ray / methods
  • Energy Transfer / immunology*
  • Epitopes, B-Lymphocyte / chemistry
  • HLA Antigens / chemistry*
  • HLA-A2 Antigen / chemistry
  • Histocompatibility Antigens Class I / chemistry*
  • Humans
  • Immunoglobulin Fab Fragments / chemistry
  • Models, Molecular*
  • Peptide Mapping
  • Receptors, Antigen, T-Cell / chemistry*
  • Spectrometry, Fluorescence / methods
  • beta 2-Microglobulin / chemistry

Substances

  • CD8 Antigens
  • Epitopes, B-Lymphocyte
  • HLA Antigens
  • HLA-A2 Antigen
  • Histocompatibility Antigens Class I
  • Immunoglobulin Fab Fragments
  • Receptors, Antigen, T-Cell
  • beta 2-Microglobulin