Modulation of complement receptors of a human monocyte cell line, U-937, during incubation with phorbol myristate acetate: expression of an iC3b-specific receptor (CR3).
The human monocyte line, U-937, derived from an individual with histiocytic lymphoma was studied for the expression of surface C3 receptors, after cultivation in the presence of phorbol myristate acetate (PMA) or T lymphocyte-conditioned medium. Receptors were detected by using EAC4b, EAC3b, EC3b, EAC3bi and EAC3d intermediates. U-937 cells, in exponential growth phase, poorly bound the intermediates; after exposure to PMA or T lymphocyte-conditioned medium, U-937 cells strongly bound both EAC3b and EAC3bi since about 50% of cells rosetted with these intermediates. This binding was totally inhibited by EDTA and by Mac-1 monoclonal antibody, suggesting the presence of only CR3 receptor types on these cells. Although U-937 cells formed rosettes with EAC3b, there was no evidence for the presence of CR1 receptors since no rosette was observed either with EAC4b or with EC3b intermediates (EC3b were prepared by coupling purified C3b to erythrocytes with N-succinimidyl 3-(2-pyridyldithio)propionate. As small amounts of factor H were present on EAC3b intermediates, incubation of EAC3b with U-937 cells induced their transformation into EAC3bi and their binding to CR3. Moreover, U-937 cells did not promote the cleavage of C3b in the presence of factor I alone, suggesting that these cells did not bear a sufficient amount of functionally active CR1. These results demonstrated that U-937 cells predominantly expressed CR3. The study of the kinetics of EAC3bi rosette formation demonstrated that CR3 expression is closely related to PMA activation. We suggest that CR3 activity could result from a phosphorylation of existing receptors.
Gilbert
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D
Peulve
P
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Daveau
M
M
Ripoche
J
J
Fontaine
M
M
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008264:Macrophages / Q000276:immunology; D009000:Monocytes / Q000276:immunology*; D010704:Phorbols / Q000494:pharmacology*; D011951:Receptors, Complement / Q000187:drug effects* / Q000378:metabolism; D017463:Receptors, Complement 3b; D012397:Rosette Formation; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D003470:Culture Media; D010704:Phorbols; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D004492:Edetic Acid; D013755:Tetradecanoylphorbol Acetate