The inhibitory function of CTLA-4 does not require its tyrosine phosphorylation

J Immunol. 2000 Jan 1;164(1):49-55. doi: 10.4049/jimmunol.164.1.49.

Abstract

CTLA-4 is a negative regulator of T cell responses. Sequence analysis of this molecule reveals the presence of two cytoplasmic tyrosine residues at positions 165 and 182 that are potential Src homology (SH)-2 domain binding sites. The role of phosphorylation of these residues in CTLA-4-mediated signaling is unknown. Here, we show that sole TCR ligation induces zeta-associated protein (ZAP)-70-dependent tyrosine phosphorylation of CTLA-4 that is important for cell surface retention of this molecule. However, CTLA-4 tyrosine phosphorylation is not required for down-regulation of T cell activation following CD3-CTLA-4 coengagement. Specifically, inhibition of extracellular signal-regulated kinase (ERK) activation and of IL-2 production by CTLA-4-mediated signaling occurs in T cells expressing mutant CTLA-4 molecules lacking the cytoplasmic tyrosine residues, and in lck-deficient or ZAP-70-deficient T cells. Therefore, CTLA-4 function involves interplay between two different levels of regulation: phosphotyrosine-dependent cell surface retention and phosphotyrosine-independent association with signaling molecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abatacept
  • Antigens, CD
  • Antigens, Differentiation / biosynthesis
  • Antigens, Differentiation / genetics
  • Antigens, Differentiation / metabolism*
  • Antigens, Differentiation / physiology*
  • CD3 Complex / immunology
  • CD3 Complex / metabolism
  • CTLA-4 Antigen
  • Cell Membrane / immunology
  • Cell Membrane / metabolism
  • Cytoplasm / immunology
  • Cytoplasm / metabolism
  • Enzyme Activation / immunology
  • Humans
  • Immunoconjugates*
  • Immunosuppressive Agents / metabolism*
  • Interleukin-2 / antagonists & inhibitors
  • Interleukin-2 / biosynthesis
  • Jurkat Cells
  • Ligands
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / physiology
  • Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism
  • Mutagenesis, Site-Directed
  • Phosphorylation
  • Protein-Tyrosine Kinases / physiology
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • Tyrosine / metabolism*
  • ZAP-70 Protein-Tyrosine Kinase

Substances

  • Antigens, CD
  • Antigens, Differentiation
  • CD3 Complex
  • CTLA-4 Antigen
  • CTLA4 protein, human
  • Immunoconjugates
  • Immunosuppressive Agents
  • Interleukin-2
  • Ligands
  • Tyrosine
  • Abatacept
  • Protein-Tyrosine Kinases
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
  • ZAP-70 Protein-Tyrosine Kinase
  • ZAP70 protein, human
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases