Defective stimulus-response coupling in human monocytes infected with Leishmania donovani is associated with altered activation and translocation of protein kinase C.
Stimulus-response coupling through protein kinase C (PKC) was shown to be defective in mononuclear phagocytes (M phi) infected with Leishmania donovani. Phorbol 12-myristate 13-acetate (PMA)-induced oxidative burst activity and protein phosphorylation were markedly attenuated in infected M phi. These results were not explained either by quantitative alterations in amounts of PKC or by altered phorbol ester binding but were related to defects in kinase activation. Analysis in vitro of the kinetic properties of PKC from infected M phi revealed an approximately 2-fold increase in the concentration of 1,2-dioleoyl-rac-glycerol required to achieve half-maximal kinase activation. Evidence for abnormal PKC activation in vivo was reflected by attenuation of PMA-induced translocation of enzyme to the particulate fraction of infected cells. These results provide direct evidence that infection with Leishmania inhibits activation of, and therefore intracellular signaling dependent on, PKC. Inhibition of stimulus-response coupling through PKC provides a basis for understanding impairment of cellular activation by Leishmania and may contribute to chronic infection.
Olivier
M
M
Department of Medicine, University of British Columbia Faculty of Medicine, Vancouver, Canada.
Brownsey
R W
RW
Reiner
N E
NE
D000818:Animals; D015153:Blotting, Western; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation; D006801:Humans; D006861:Hydrogen Peroxide / Q000097:blood*; D066298:In Vitro Techniques; D007700:Kinetics; D007893:Leishmania donovani / Q000472:pathogenicity*; D009000:Monocytes / Q000187:drug effects / Q000469:parasitology / Q000502:physiology*; D010101:Oxygen Consumption / Q000187:drug effects; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D010710:Phosphates / Q000097:blood; D010750:Phosphoproteins / Q000097:blood* / Q000302:isolation & purification; D011493:Protein Kinase C / Q000097:blood*; D011955:Receptors, Drug / Q000378:metabolism; D013481:Superoxides / Q000097:blood*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D004075:Diglycerides; D010710:Phosphates; D010750:Phosphoproteins; D011955:Receptors, Drug; C083141:phorbol ester binding protein; C027776:phorbol ester receptor; D013481:Superoxides; D015240:Phorbol 12,13-Dibutyrate; D006861:Hydrogen Peroxide; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; C013965:diolein