Differential effect of the activation of protein kinase A on the protein synthesis and secretion in the T-helper 2 cell line D10.G4.1

Scand J Immunol. 1996 Aug;44(2):150-6. doi: 10.1046/j.1365-3083.1996.d01-295.x.

Abstract

The authors analysed the effect of protein kinase A (PKA) activation on the protein synthesis and secretion in the T-helper 2 cell line D10.G4.1 (D10) using an assay that allows the detection of almost all secreted proteins of a cell. IL-4 and IL-10 were quantified. Three groups of secretory products could be defined. The T-cell receptor (TCR)-induced production of the first group (A) of proteins including IL-4 was enhanced by low concentrations of PKA activators. At higher concentrations the enhancement was less marked. The synthesis and secretion of a second group (B) of proteins including IL-10 remained unaffected. The production of a third group (C) of proteins was inhibited in a concentration-dependent manner. Biochemical analysis revealed a block of phospholipase C gamma (PLC gamma) activity by PKA activators. When D10 cells were stimulated by a phorbol ester plus calcium ionophore the production of group A proteins was enhanced almost fourfold, whereas production of group B proteins was unaffected by PKA activation. This effect was observed at all concentrations of various PKA activators tested. The secretion of group C proteins was no longer inhibited. The same results were obtained when analysing IL-4 and IL-10 m-RNA by Northern blotting. The data demonstrate a lymphokine specific mode of action on a single cell basis. Furthermore, it suggests that the inhibitory action of PKA in D10 cells is due partly to blocking of PLC gamma activity.

MeSH terms

  • 1-Methyl-3-isobutylxanthine / pharmacology
  • Animals
  • Cell Line
  • Cyclic AMP-Dependent Protein Kinases / physiology*
  • Enzyme Activation
  • Humans
  • Interleukin-10 / biosynthesis*
  • Interleukin-10 / genetics
  • Interleukin-10 / metabolism
  • Interleukin-4 / biosynthesis*
  • Interleukin-4 / genetics
  • Interleukin-4 / metabolism
  • Mice
  • Mice, Inbred C3H
  • RNA, Messenger / analysis
  • Rabbits
  • Rats
  • Receptors, Antigen, T-Cell / physiology
  • Tetradecanoylphorbol Acetate / pharmacology
  • Th2 Cells / metabolism
  • Type C Phospholipases / antagonists & inhibitors

Substances

  • RNA, Messenger
  • Receptors, Antigen, T-Cell
  • Interleukin-10
  • Interleukin-4
  • Cyclic AMP-Dependent Protein Kinases
  • Type C Phospholipases
  • Tetradecanoylphorbol Acetate
  • 1-Methyl-3-isobutylxanthine