Involvement of protein kinase C in activation of human granulocytes and peritoneal macrophages by type 1 fimbriated (mannose specific) Escherichia coli.
Specific binding of bacteria to phagocytic cells mediated by antibody and complement (opsonins) or by lectin-carbohydrate interactions is required for their efficient uptake and killing by opsonophagocytosis or lectinophagocytosis, respectively (Ofek and Sharon, Infect. Immun. 56, 539, 1988). An early step in these processes is activation of the phagocytes by the bound bacteria, as evidenced by appearance of an oxidative burst. Previous work has shown that protein kinase C (PKC) is involved in activation of human granulocytes by opsonized Escherichia coli. In the present study, we used three inhibitors of PKC to examine the possible involvement of the enzyme in activation of human granulocytes and peritoneal macrophages by type 1 fimbriated (mannose-specific) Escherichia coli in the absence of opsonins. Activation, as measured by chemiluminescence, was completely inhibited by sphingosine (50 microM) and only partially (50%) by 100 microM H-7 [1-(5-isoquinolinesulfonyl)-2-methylpiperazine]; in both cases it was fully reversible. The third inhibitor, K252a, also inhibited almost completely the activation at 1 microM. The inhibitors acted similarly on activation of the phagocytic cells by opsonized bacteria or by phorbol-12-myristate-13-acetate (0.1 microM). Down regulation of the kinase, by pretreatment of the human granulocytes or macrophages with a high concentration (1.6 microM) of phorbol myristate acetate, abolished their ability to respond to stimulation by the bacteria. Our findings provide evidence for the involvement of PKC in the activation of phagocytic cells by type 1 fimbriated bacteria.
Gbarah
A
A
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Mhashilkar
A M
AM
Boner
G
G
Sharon
N
N
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D002227:Carbazoles / Q000494:pharmacology; D004926:Escherichia coli / Q000502:physiology*; D010861:Fimbriae, Bacterial / Q000502:physiology*; D006098:Granulocytes / Q000502:physiology*; D026121:Indole Alkaloids; D007546:Isoquinolines / Q000494:pharmacology; D008163:Luminescent Measurements; D008262:Macrophage Activation / Q000502:physiology*; D008264:Macrophages / Q000502:physiology*; D008358:Mannose; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D013110:Sphingosine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002227:Carbazoles; D026121:Indole Alkaloids; D007546:Isoquinolines; D010879:Piperazines; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C049985:staurosporine aglycone; D011493:Protein Kinase C; D013110:Sphingosine; D013755:Tetradecanoylphorbol Acetate; D008358:Mannose