JAK3, STAT, and MAPK signaling pathways as novel molecular targets for the tyrphostin AG-490 regulation of IL-2-mediated T cell response

J Immunol. 1999 Apr 1;162(7):3897-904.

Abstract

AG-490 is a member of the tyrphostin family of tyrosine kinase inhibitors. While AG-490 has been considered to be a Janus kinase (JAK)2-specific inhibitor, these conclusions were primarily drawn from acute lymphoblastic leukemia cells that lack readily detectable levels of JAK3. In the present study, evidence is provided that clearly demonstrates AG-490 potently suppresses IL-2-induced T cell proliferation, a non-JAK2-dependent signal, in a dose-dependent manner in T cell lines D10 and CTLL-2. AG-490 blocked JAK3 activation and phosphorylation of its downstream counterpart substrates, STATs. Inhibition of JAK3 by AG-490 also compromised the Shc/Ras/Raf/mitogen-activated protein kinase (MAPK) signaling pathways as measured by phosphorylation of Shc and extracellular signal-related kinase 1 and 2 (ERK1/2). AG-490 effectively inhibited tyrosine phosphorylation and DNA binding activities of several transcription factors including STAT1, -3, -5a, and -5b and activating protein-1 (AP-1) as judged by Western blot analysis and electrophoretic mobility shift assay. These data suggest that AG-490 is a potent inhibitor of the JAK3/STAT, JAK3/AP-1, and JAK3/MAPK pathways and their cellular consequences. Taken together, these findings support the notion that AG-490 possesses previously unrecognized clinical potential as an immunotherapeutic drug due to its inhibitory effects on T cell-derived signaling pathways.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Adaptor Proteins, Vesicular Transport*
  • Binding Sites / drug effects
  • Binding Sites / immunology
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Cell Line
  • DNA / metabolism
  • DNA-Binding Proteins / antagonists & inhibitors
  • DNA-Binding Proteins / metabolism*
  • Enzyme Activation / drug effects
  • Enzyme Activation / immunology
  • Enzyme Inhibitors / pharmacology
  • Interleukin-2 / antagonists & inhibitors
  • Interleukin-2 / pharmacology
  • Janus Kinase 3
  • Lymphocyte Activation / drug effects
  • Milk Proteins*
  • Phosphorylation / drug effects
  • Phosphotyrosine / metabolism
  • Protein-Tyrosine Kinases / antagonists & inhibitors
  • Protein-Tyrosine Kinases / metabolism*
  • Proteins / metabolism
  • RNA, Messenger / metabolism
  • Receptors, Interleukin-2 / biosynthesis
  • STAT1 Transcription Factor
  • STAT3 Transcription Factor
  • STAT5 Transcription Factor
  • Shc Signaling Adaptor Proteins
  • Signal Transduction / drug effects
  • Signal Transduction / immunology*
  • T-Lymphocytes / drug effects*
  • T-Lymphocytes / enzymology*
  • T-Lymphocytes / immunology
  • Trans-Activators / antagonists & inhibitors
  • Trans-Activators / metabolism*
  • Tyrphostins / pharmacology*

Substances

  • Adaptor Proteins, Signal Transducing
  • Adaptor Proteins, Vesicular Transport
  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Interleukin-2
  • Milk Proteins
  • Proteins
  • RNA, Messenger
  • Receptors, Interleukin-2
  • STAT1 Transcription Factor
  • STAT3 Transcription Factor
  • STAT5 Transcription Factor
  • Shc Signaling Adaptor Proteins
  • Trans-Activators
  • Tyrphostins
  • alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide
  • Phosphotyrosine
  • DNA
  • Protein-Tyrosine Kinases
  • Janus Kinase 3
  • Calcium-Calmodulin-Dependent Protein Kinases