Effect of 1,25(OH)(2)-vitamin D(3) on the activation of natural killer cells: role of protein kinase C and extracellular calcium

Exp Mol Pathol. 1999 Oct;67(2):63-74. doi: 10.1006/exmp.1999.2264.

Abstract

As a first approach for studying the implication of PKC and the steroid hormone 1,25(OH)(2)-vitamin D(3) [1,25(OH)(2)D(3)] on natural killer cell (NK) activity, we analyzed in the YT NK cell line the expression of PKC isoforms and the effects of 1, 25(OH)(2)D(3) on BLT-esterase (a marker of NK lytic granules) activity. Western blot and RT-PCR showed a greater extent of PKC alpha, beta, delta, zeta, epsilon, theta, and lambda and lower levels of PKC mu and eta. In a dose-dependent manner 1, 25(OH)(2)D(3) induced significant increases in BLT-esterase and PKC activities and the stimulatory effect on BLT-esterase activity was mimicked and blocked, respectively, by the PKC activator phorbol ester PMA and PKC inhibitors (H7, PKC(19-36), and N-myristoylated PKC(19-31) peptides). Moreover, the effects of 1,25(OH)(2)D(3) on BLT-esterase could be blocked in a Ca(2+)-free (+EGTA) medium and mimicked by the Ca2+ ionophore A23187. The results suggest that 1, 25(OH)(2)D(3) is a stimulatory factor of NK activity acting through a mechanism involving PKC and extracellular Ca2+.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Calcimycin / pharmacology
  • Calcitriol / pharmacology*
  • Calcium / metabolism*
  • Cell Line
  • DNA Primers / genetics
  • Egtazic Acid / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Extracellular Space / metabolism
  • Gene Expression / drug effects
  • Granzymes
  • Humans
  • Ionophores / pharmacology
  • Isoenzymes / antagonists & inhibitors
  • Isoenzymes / genetics
  • Isoenzymes / metabolism
  • Killer Cells, Natural / drug effects*
  • Killer Cells, Natural / immunology
  • Killer Cells, Natural / metabolism*
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / genetics
  • Protein Kinase C / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Serine Endopeptidases / metabolism
  • Subcellular Fractions / enzymology
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • DNA Primers
  • Enzyme Inhibitors
  • Ionophores
  • Isoenzymes
  • RNA, Messenger
  • Calcimycin
  • Egtazic Acid
  • Protein Kinase C
  • Granzymes
  • Serine Endopeptidases
  • GZMA protein, human
  • Calcitriol
  • Tetradecanoylphorbol Acetate
  • Calcium