Fluorosomes: a convenient new reagent to detect and block multivalent and complex receptor-ligand interactions

FASEB J. 2010 May;24(5):1572-82. doi: 10.1096/fj.09-137281. Epub 2010 Jan 7.

Abstract

We describe for the first time fluorescent virus-like particles decorated with biologically active mono- and multisubunit immune receptors of choice and the basic application of such fluorosomes (FSs) to visualize and target immune receptor-ligand interactions. For that purpose, human embryonic kidney (HEK)-293 cells were stably transfected with Moloney murine leukemia virus (MoMLV) matrix protein (MA) GFP fusion constructs. To produce FSs, interleukins (ILs), IL-receptors (IL-Rs), and costimulatory molecules were fused to the glycosyl phosphatidyl inositol anchor acceptor sequence of CD16b and coexpressed along with MoMLV group-specific antigen-polymerase (gag-pol) in MA::GFP(+) HEK-293 cells. We show that IL-2 decorated but not control-decorated FSs specifically identify normal and malignant IL-2 receptor-positive (IL-2R(+)) lymphocytes by flow cytometry. In addition to cytokines and costimulatory molecules, FSs were also successfully decorated with the heterotrimeric IL-2Rs, allowing identification of IL-2(+) target cells. Specificity of binding was proven by complete inhibition with nonlabeled, soluble ligands. Moreover, IL-2R FSs efficiently neutralized soluble IL-2 and thus induced unresponsiveness of T cells receiving full activation stimuli via T-cell antigen receptor and CD28. FSs are technically simple, multivalent tools for assessing and blocking mono- and multisubunit immune receptor-ligand interactions with natural constituents in a plasma membrane context.

Publication types

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

MeSH terms

  • Cell Line, Transformed
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • GPI-Linked Proteins
  • Green Fluorescent Proteins / chemistry*
  • Green Fluorescent Proteins / genetics
  • Humans
  • Interleukin-2 / analysis
  • Interleukin-2 / immunology*
  • Ligands
  • Lymphoma, B-Cell / diagnosis
  • Lymphoma, B-Cell / immunology
  • Microscopy, Confocal
  • Moloney murine leukemia virus
  • Receptors, IgG
  • Receptors, Interleukin-2 / analysis
  • Receptors, Interleukin-2 / immunology*
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / genetics
  • T-Lymphocytes / immunology
  • Virion / chemistry*
  • Virion / genetics

Substances

  • FCGR3B protein, human
  • Fluorescent Dyes
  • GPI-Linked Proteins
  • Interleukin-2
  • Ligands
  • Receptors, IgG
  • Receptors, Interleukin-2
  • Recombinant Proteins
  • Green Fluorescent Proteins