Tumor-promoting phorbol esters induce rapid internalization of the C3b receptor via a cytoskeleton-dependent mechanism.
Plasma membrane expression as well as phagocytic capability of the C3b receptor (CR1) are under regulatory control. Phorbol esters are one class of agents which have been shown to influence both of these events. In this study, by using radiolabeled Fab fragments of a monoclonal anti-CR1 antibody to tag the receptor and acid elution of surface-bound Fab, we showed that both phorbol myristate acetate and phorbol dibutyrate induced internalization of the C3b receptor; this occurred in a dose- and time-dependent manner in the absence of occupancy of the receptor by ligand. This was shown to occur in neutrophils, monocytes, and macrophages. We also showed that phorbol esters enhanced CR1-dependent phagocytosis despite the presence of two-thirds fewer receptors present on the plasma membrane. However, fibronectin, another agent that influences phagocytosis, had no effect on receptor internalization. Phorbol ester internalization was temperature-dependent and was inhibitable by cytochalasins B and D. Inhibition of internalization was reversible when cytochalasin B was removed. Phorbol esters also induced increased detergent insolubility of CR1 with kinetics similar to those of receptor internalization. It is possible that association of CR1 with the cytoskeleton is important to the process of "activation" of CR1 in phagocytosis.
O'Shea
J J
JJ
Brown
E J
EJ
Gaither
T A
TA
Takahashi
T
T
Frank
M M
MM
D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism*; D005353:Fibronectins / Q000494:pharmacology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism; D010587:Phagocytosis / Q000187:drug effects*; D015240:Phorbol 12,13-Dibutyrate; D010703:Phorbol Esters / Q000494:pharmacology*; D010704:Phorbols / Q000494:pharmacology*; D011951:Receptors, Complement / Q000187:drug effects* / Q000276:immunology; D017463:Receptors, Complement 3b; D013696:Temperature; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D007140:Immunoglobulin Fab Fragments; D010703:Phorbol Esters; D010704:Phorbols; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D015240:Phorbol 12,13-Dibutyrate; D003571:Cytochalasin B; D013755:Tetradecanoylphorbol Acetate