Supramolecular receptor structures in the plasma membrane of lymphocytes revealed by flow cytometric energy transfer, scanning force- and transmission electron-microscopic analyses

Cytometry. 1998 Oct 1;33(2):225-33.

Abstract

Receptors in the plasma membrane of blood cells in general and in that of lymphocytes in particular are supposed to move around in a random walk fashion relatively freely driven by thermal diffusion, as described by the Singer-Nicolson fluid mosaic membrane model. In this article we summarized data and techniques that indicated nonrandom codistribution patterns of receptor superstructures under conditions, where the generation of such molecular colocalizations by the methods themselves were excluded. Application of fluorescence energy transfer in a flow cytometer helped to analyze such codistribution patterns in cell populations. After normalizing energy transfer values for possible differences between labeling ratios of the targeting monoclonal antibodies and using the mean values of energy transfer distribution curves, two-dimensional receptor maps were generated from data obtained in a pair-wise fashion between receptors. Major histocompatibility complex (MHC) class I and II, intercellular adhesion molecule-1 (ICAM-1), TcR-CD3-CD4, tetraspan molecules (CD81, CD82, CD53), and the subunits of the multisubunit IL-2 receptor displayed nonrandom codistribution patterns sometimes with, but very frequently without induction by their ligand. Immunogold-bead "sandwich" labeling analyzed by atomic force microscopy has shown that such receptor "islands" existed also in "receptor-island-groups". This indicated the existence of nonrandom receptor distribution of MHC class I and II molecules also at an elevated hierarchical level. An analysis is given herein concerning a standardized approach. The apparent incompatibility of these supramolecular patterns with the Singer-Nicolson type "free-protein and lipid-mobility paradigm" was resolved by recommending an additional emphasis on the mosaicism of the membrane besides receptor mobility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Antigens, CD / analysis
  • Diffusion
  • Energy Transfer
  • Flow Cytometry*
  • Fluorescence
  • Fluorescent Dyes
  • HLA Antigens / analysis
  • Humans
  • Immunoglobulin Fab Fragments / immunology
  • Immunohistochemistry
  • Intercellular Adhesion Molecule-1 / analysis
  • Lymphocytes / chemistry*
  • Macromolecular Substances
  • Membrane Fluidity
  • Microscopy, Atomic Force*
  • Microscopy, Electron*
  • Microspheres
  • Models, Biological
  • Motion
  • Receptor-CD3 Complex, Antigen, T-Cell / analysis
  • Receptors, Cell Surface / analysis*
  • Receptors, Interleukin-2 / analysis
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Fluorescent Dyes
  • HLA Antigens
  • Immunoglobulin Fab Fragments
  • Macromolecular Substances
  • Receptor-CD3 Complex, Antigen, T-Cell
  • Receptors, Cell Surface
  • Receptors, Interleukin-2
  • Intercellular Adhesion Molecule-1