Enhancement of protein kinase C in murine lymphokine-activated killer cells by retinoic acid.
We previously reported that retinoic acid (RA) augmented mouse (BALB/c) lymphokine (interleukin-2)-activated killer (LAK) cell activity in a dose and time dependent manner. As evidence available has suggested the role of protein kinase C (PKC) in the regulation of cell mediated cytotoxicity, the present work was to investigate whether or not PKC may mediate the enhancement of LAK cell activity by RA. Accompanied with an augmented LAK cell activity, RA increased total PKC enzyme activity, [3H]phorbol 12,13-dibutyrate binding activity, and the amount of immunoreactive PKC. A prolonged treatment (18 h) of LAK cells with 12-O-tetradecanoylphorbol-13-acetate resulted in the loss of both PKC and LAK cell activity. PKC inhibitors, 1-(5-isoquinolinesulfonyl)-2-methyl-piperazine dihydrochloride and staurosporine, also drastically reduced LAK cell activity. Although most of the total PKC activity (97%) was detected in the cytosol fraction, the increase in PKC activity was attributed to an increased enzyme activity in both cytosol and membrane fractions, and shown to be RA dose-dependent. Kinetics study revealed that the increase in PKC was a time-dependent process and the enhancement was detectable as early as 8 h after the addition of RA to LAK cell culture. By immunoblotting, the cytosol PKC of LAK cells was shown to contain alpha and beta isoforms, but not gamma. RA further increased the expression of PKC alpha. The enhanced expression of alpha isozyme of PKC by RA was also in a dose and time dependent manner. Taken together, these results indicate that the mechanism of the augmentation of LAK cell activity by RA may in part result from the increase in PKC, especially PKC alpha isozyme.
Lin
T H
TH
Department of Diagnostic Immunology Research and Biochemistry, Roswell Park Memorial Institute, New York State Department of Health, Buffalo 14263.
Chu
T M
TM
D000818:Animals; D015153:Blotting, Western; D003602:Cytotoxicity, Immunologic; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015979:Killer Cells, Lymphokine-Activated / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology*
D007527:Isoenzymes; D014212:Tretinoin; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate