Interleukin 10 induced c-fos expression in human B cells by activation of divergent protein kinases

Immunol Invest. 1996 Jan-Mar;25(1-2):115-28. doi: 10.3109/08820139609059296.

Abstract

IL-10 is a potent mediator of human B cell growth and plasma cell formation. However, signal transduction of IL-10 in B cells is poorly understood. In this study the effect of IL-10 on the expression of the protooncogene c-fos was investigated, because Fos plays a potential role in the regulation of B cell proliferation and differentiation. B cells were purified from buffy coat preparations of healthy blood donors by positive selection using an anti CD20 monoclonal antibody and a MiniMACS separation unit. B cells were prestimulated with SAC for 48 hrs. Then, cells were incubated with medium or IL-10 (100 ng/ml) for 10 to 120 min. RNA was extracted by phenol/chloroform and c-fos expression was analyzed by PCR assisted mRNA assay. A significant 2-4 fold increase of c-fos expression was observed within 30 min of stimulation with IL-10 (p < 0.01). After 2 hrs c-fos expression declined to basal levels. The effect of IL-10 was dose-dependent with a maximum stimulation using 100 ng/ml of IL-10. The IL-10 effect on c-fos expression was not blocked by polymyxin B. Using the tyrosine kinase inhibitor genistein (10 microM) a complete inhibition of IL-10 induced c-fos expression was observed. In addition, H-7 (10 microM), a specific inhibitor of serine/threonine kinases, significantly blocked IL-10 mediated c-fos expression (p < 0.05). In conclusion, these data show that IL-10 induces c-fos expression in human B-cells by activation of tyrosine and serine/threonine kinases. Since this is the first report on IL-10 induced signal transduction, these data may help to identify the intracellular mechanisms by which IL-10 stimulates human B-cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • B-Lymphocytes / drug effects*
  • B-Lymphocytes / metabolism
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Gene Expression Regulation / drug effects*
  • Genes, fos / drug effects*
  • Genistein
  • Humans
  • Interleukin-10 / pharmacology*
  • Isoflavones / pharmacology
  • Isoquinolines / pharmacology
  • Piperazines / pharmacology
  • Polymerase Chain Reaction
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / metabolism*
  • Protein-Tyrosine Kinases / antagonists & inhibitors
  • Protein-Tyrosine Kinases / metabolism*
  • Proto-Oncogene Proteins c-fos / biosynthesis*
  • Proto-Oncogene Proteins c-fos / physiology
  • Signal Transduction / drug effects*
  • Signal Transduction / physiology

Substances

  • Enzyme Inhibitors
  • Isoflavones
  • Isoquinolines
  • Piperazines
  • Proto-Oncogene Proteins c-fos
  • Interleukin-10
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Genistein
  • Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases