Immunoglobulin-mediated phagocytosis by human monocytes requires protein kinase C activation. Evidence for protein kinase C translocation to phagosomes.
This study has investigated the role of protein kinase C (PKC) activation in IgG-mediated phagocytosis by human monocytes. Incubation of monocytes with IgG-opsonized targets increased membrane-associated PKC approximately 2-fold. Kinetic studies showed that the translocation of PKC to membrane occurred before significant ingestion took place. The pharmacologic PKC inhibitor H7 inhibited IgG-dependent ingestion with ID50 of 20 microM, while the structurally related isoquinoline sulfonamide HA1004 had no effect at this concentration. Staurosporine and calphostin C, PKC inhibitors which have different mechanisms of actions than H7, also inhibited ingestion. Depletion of PKC by prolonged incubation with phorbol esters also inhibited phagocytosis, and dose-response curves showed a strong correlation between the extent of PKC depletion and the extent of inhibition of ingestion. Finally, phagosomes were isolated by sucrose density centrifugation of cells disrupted 5 min after the initiation of phagocytosis. Measurement of PKC activity and immunoreactivity in the phagosomes showed that PKC was concentrated in the phagosome membrane approximately 5-fold compared to the uninvolved plasma membrane. Together, these data suggest that PKC activation is an early, essential step in the efficient ingestion of IgG-opsonized targets by monocytes.
Zheleznyak
A
A
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Brown
E J
EJ
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000469:Alkaline Phosphatase / Q000378:metabolism; D000470:Alkaloids / Q000494:pharmacology; D000595:Amino Acid Sequence; D001692:Biological Transport; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005136:Eye Proteins; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D007546:Isoquinolines / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D009000:Monocytes / Q000276:immunology*; D009281:Naphthalenes; D010587:Phagocytosis; D010588:Phagosomes / Q000201:enzymology*; D010879:Piperazines / Q000494:pharmacology; D011083:Polycyclic Compounds / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D017299:Rod Opsins; D015398:Signal Transduction; D019311:Staurosporine; D013449:Sulfonamides
D000470:Alkaloids; D005136:Eye Proteins; D007074:Immunoglobulin G; D007546:Isoquinolines; D009281:Naphthalenes; D010879:Piperazines; D011083:Polycyclic Compounds; D017299:Rod Opsins; D013449:Sulfonamides; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C043204:N-(2-guanidinoethyl)-5-isoquinolinesulfonamide; D011493:Protein Kinase C; D000469:Alkaline Phosphatase; D019311:Staurosporine; C058819:calphostin C