Effect of 1,25(OH)(2)-vitamin D(3) on the activation of natural killer cells: role of protein kinase C and extracellular calcium.
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+.
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Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, Bahía Blanca, 8000, Argentina.
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Boland
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Barja
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D001483:Base Sequence; D000001:Calcimycin / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004533:Egtazic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005110:Extracellular Space / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D053804:Granzymes; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D017931:DNA Primers; D004791:Enzyme Inhibitors; D007476:Ionophores; D007527:Isoenzymes; D012333:RNA, Messenger; D000001:Calcimycin; D004533:Egtazic Acid; D011493:Protein Kinase C; D053804:Granzymes; D012697:Serine Endopeptidases; C509212:GZMA protein, human; D002117:Calcitriol; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium