New disease-modifying antirheumatic drug 2 acetylthiomethyl-4-(4-methylphenyl)-4-oxobutanoic acid (KE-298) selectively augments activation-induced T cell death

J Lab Clin Med. 2001 Jul;138(1):11-7. doi: 10.1067/mlc.2001.115938.

Abstract

We examined in this study whether the newly developed disease-modifying antirheumatic drug (DMARD) 2-acetylthiomethyl-4-(4-methylphenyl)-4-oxobutanoic acid (KE-298) augments activation-induced T cell death. Peripheral blood (PB) T cells, isolated from healthy donors, were activated by incubation with interleukin-2 (IL-2) followed by further culture with 12-0-tetradecanoyl phorbol 13-acetate (PMA) and ionomycin in the presence or absence of KE-298. The apoptosis of activated T cells was examined by flow cytometric determination of hypodiploid DNA. Fas expression and caspase-3 activity in activated T cells were also examined by flow cytometry, and expression of Fas ligand (FasL), Bcl-2-related proteins, and X chromosome-linked inhibitor of apoptosis protein (XIAP) was determined by Western blot analysis. Apoptosis was not obvious in resting T cells and was not augmented by KE-298. In contrast, apoptosis was clearly detected in activated T cells (activation-induced T cell death) with the increment of caspase-3 activity, and incubation of these cells with KE-298 further enhanced apoptosis. Treatment of activated T cells with KE-298 increased Bax expression but decreased XIAP expression without affecting the expression of Fas/FasL. Thus caspase-3 activity in activated T cells appeared to be increased by KE-298. Our results suggest that the newly developed DMARD, KE-298, selectively augmented activation-induced T cell death. This finding may contribute to the therapeutic efficacy of KE-298 in rheumatoid arthritis (RA) patients and provide new insight into the pharmacologic action of DMARDs.

MeSH terms

  • Antirheumatic Agents / pharmacology*
  • Apoptosis / drug effects*
  • Apoptosis / immunology
  • Blotting, Western
  • Caspase 3
  • Caspases / metabolism
  • Cells, Cultured
  • Cysteine Proteinase Inhibitors / pharmacology
  • Fas Ligand Protein
  • Flow Cytometry
  • Humans
  • Lymphocyte Activation / drug effects
  • Lymphocyte Activation / immunology
  • Membrane Glycoproteins / analysis
  • Membrane Glycoproteins / metabolism
  • Oligopeptides / pharmacology
  • Phenylpropionates / pharmacology*
  • Proteins / analysis
  • Proteins / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / analysis
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • T-Lymphocytes / chemistry
  • T-Lymphocytes / cytology*
  • T-Lymphocytes / enzymology
  • X-Linked Inhibitor of Apoptosis Protein
  • bcl-X Protein
  • fas Receptor / analysis
  • fas Receptor / metabolism

Substances

  • Antirheumatic Agents
  • BCL2L1 protein, human
  • Cysteine Proteinase Inhibitors
  • FASLG protein, human
  • Fas Ligand Protein
  • Membrane Glycoproteins
  • Oligopeptides
  • Phenylpropionates
  • Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • X-Linked Inhibitor of Apoptosis Protein
  • XIAP protein, human
  • aspartyl-glutamyl-valyl-aspartal
  • bcl-X Protein
  • fas Receptor
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • esonarimod