Regulation of CD93 cell surface expression by protein kinase C isoenzymes.
Human CD93, also known as complement protein 1, q subcomponent, receptor (C1qRp), is selectively expressed by cells with a myeloid lineage, endothelial cells, platelets, and microglia and was originally reported to be involved in the complement protein 1, q subcomponent (C1q)-mediated enhancement of phagocytosis. The intracellular molecular events responsible for the regulation of its expression on the cell surface, however, have not been determined. In this study, the effect of protein kinases in the regulation of CD93 expression on the cell surface of a human monocyte-like cell line (U937), a human NK-like cell line (KHYG-1), and a human umbilical vein endothelial cell line (HUV-EC-C) was investigated using four types of protein kinase inhibitors, the classical protein kinase C (cPKC) inhibitor Go6976, the novel PKC (nPKC) inhibitor Rottlerin, the protein kinase A (PKA) inhibitor H-89 and the protein tyrosine kinase (PTK) inhibitor herbimycin A at their optimum concentrations for 24 hr. CD93 expression was analyzed using flow cytometry and glutaraldehyde-fixed cellular enzyme-linked immunoassay (EIA) techniques utilizing a CD93 monoclonal antibody (mAb), mNI-11, that was originally established in our laboratory as a CD93 detection probe. The nPKC inhibitor Rottlerin strongly down-regulated CD93 expression on the U937 cells in a dose-dependent manner, whereas the other inhibitors had little or no effect. CD93 expression was down-regulated by Go6976, but not by Rottlerin, in the KHYG-1 cells and by both Rottlerin and Go6976 in the HUV-EC-C cells. The PKC stimulator, phorbol myristate acetate (PMA), strongly up-regulated CD93 expression on the cell surface of all three cell-lines and induced interleukin-8 (IL-8) production by the U937 cells and interferon-gamma (IFN-gamma) production by the KHYG-1 cells. In addition, both Go6976 and Rottlerin inhibited the up-regulation of CD93 expression induced by PMA and IL-8 or IFN-gamma production in the respective cell-lines. Whereas recombinant tumor necrosis factor-alpha (rTNF-alpha) slightly up-regulated CD93 expression on the U937 cells, recombinant interleukin-1beta (rIL-1beta), recombinant interleukin-2 (rIL-2), recombinant interferon-gamma (rIFN-gamma) and lipopolysaccharide (LPS) had no effect. Taken together, these findings indicate that the regulation of CD93 expression on these cells involves the PKC isoenzymes.
Ikewaki
Nobunao
N
Institute of Immunology, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan.
Kulski
Jerzy K
JK
Inoko
Hidetoshi
H
D000098:Acetophenones / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology; D016227:Benzoquinones; D002227:Carbazoles / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects; D042783:Endothelial Cells / Q000201:enzymology / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D007124:Immunoenzyme Techniques; D007211:Indoles / Q000494:pharmacology; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology; D016209:Interleukin-8 / Q000032:analysis / Q000276:immunology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology; D047029:Lactams, Macrocyclic; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000276:immunology; D009000:Monocytes / Q000201:enzymology / Q000276:immunology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011809:Quinones / Q000494:pharmacology; D011951:Receptors, Complement / Q000096:biosynthesis* / Q000276:immunology; D017828:Rifabutin / Q000031:analogs & derivatives; D013449:Sulfonamides / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D020298:U937 Cells; D015854:Up-Regulation / Q000187:drug effects
D000098:Acetophenones; D001578:Benzopyrans; D016227:Benzoquinones; D002227:Carbazoles; D007211:Indoles; D016209:Interleukin-8; D007527:Isoenzymes; D007546:Isoquinolines; D047029:Lactams, Macrocyclic; D008562:Membrane Glycoproteins; D047428:Protein Kinase Inhibitors; D011809:Quinones; D011951:Receptors, Complement; D013449:Sulfonamides; C032402:complement 1q receptor; C081021:Go 6976; D017828:Rifabutin; C020754:herbimycin; D007371:Interferon-gamma; C085746:rottlerin; D011493:Protein Kinase C; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D013755:Tetradecanoylphorbol Acetate