In vivo disruption of T cell development by expression of a dominant-negative polypeptide designed to abolish the SLP-76/Gads interaction

Eur J Immunol. 2007 Oct;37(10):2961-72. doi: 10.1002/eji.200636855.

Abstract

Multi-molecular complexes nucleated by adaptor proteins play a central role in signal transduction. In T cells, one central axis consists of the assembly of several signaling proteins linked together by the adaptors linker of activated T cells (LAT), Src homology 2 domain-containing leukocyte-specific phosphoprotein of 76 kDa (SLP-76), and Grb2-related adaptor downstream of Shc (Gads). Each of these adaptors has been shown to be important for normal T cell development, and their proper sub-cellular localization is critical for optimal function in cell lines. We previously demonstrated in Jurkat T cells and a rat basophilic leukemic cell line that expression of a 50-amino acid polypeptide identical to the site on SLP-76 that binds to Gads blocks proper localization of SLP-76 and SLP-76-dependent signaling events. Here we extend these studies to investigate the ability of this polypeptide to inhibit TCR-induced integrin activity in Jurkat cells and to inhibit in vivo thymocyte development and primary T cell function. These data provide evidence for the in vivo function of a dominant-negative peptide based upon the biology of SLP-76 action and suggest the possibility of therapeutic potential of targeting the SLP-76/Gads interaction.

Publication types

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

MeSH terms

  • Adaptor Proteins, Signal Transducing / antagonists & inhibitors
  • Adaptor Proteins, Signal Transducing / deficiency
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Animals
  • Cell Adhesion / immunology
  • Cell Differentiation / immunology*
  • Cell Line, Tumor
  • Cell Movement / immunology
  • Cells, Cultured
  • Chemokine CXCL12 / physiology
  • Growth Inhibitors / chemical synthesis
  • Growth Inhibitors / genetics
  • Growth Inhibitors / physiology*
  • Humans
  • Integrins / antagonists & inhibitors
  • Integrins / physiology
  • Jurkat Cells
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Peptide Fragments / biosynthesis
  • Peptide Fragments / genetics
  • Peptide Fragments / physiology
  • Peptides / chemical synthesis*
  • Peptides / genetics*
  • Peptides / physiology
  • Phosphoproteins / antagonists & inhibitors
  • Phosphoproteins / metabolism*
  • Protein Binding / immunology
  • Rats
  • T-Lymphocytes / cytology*
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • CXCL12 protein, human
  • Chemokine CXCL12
  • Growth Inhibitors
  • Integrins
  • Mona protein, mouse
  • Peptide Fragments
  • Peptides
  • Phosphoproteins
  • SLP-76 signal Transducing adaptor proteins