Characterization of the human neutrophil C1q receptor and functional effects of free ligand on activated neutrophils.
The partial characterization and expression of the C1q receptor (C1q-R) in relation to other complement receptors present on the surface of neutrophils has been examined, as well as the effects of free C1q on cell function. A polyclonal anti-C1q-R antibody recognizes a 68-kD neutrophil surface protein. C1q-R expression was not upregulated upon warming, priming, or exposure to FMLP, but decreased after exposure to phorbol myristate acetate (PMA), because of shedding of the receptor into the extracellular medium, as detected by enzyme-linked immunosorbent assay. CR3 and CR1 expression was upregulated from intracellular pools after cell stimulation by PMA. No evidence of intracellular pools of C1q-R was found, as assessed by immunoblotting of subcellular fractions. But C1q-R appeared to be expressed early in cell differentiation, was detected on undifferentiated HL-60 cells, and like CR3 expression, increased upon 5 days differentiation towards a neutrophil lineage. However, C1q-R expression decreased upon additional culture, whereas CR3 expression continued to increase. A large variation in the percentage of peripheral cells expressing C1q receptors in donors was observed, ranging from 13% to 100%, contrasting with CR3 receptors that exhibited less variability. Interactions between free monomeric C1q and neutrophils were also studied. Incubation of stimulated neutrophils with 10 to 100 micrograms/mL C1q resulted in a further increase in CR3 expression and adherence to albumin-coated surfaces. Staphylococci opsonized with low quantities of C1q (0.1 to 1 microgram/mL) mediated a moderate and sustained respiratory burst in neutrophils, whereas a burst of similar magnitude was generated only with free C1q at concentrations 10- to 100-fold higher. Stimulation was only partially inhibited if cells were first treated with anti-C1q-R antibody, suggesting other C1q binding proteins may be present on the cell surface. In summary, neutrophil C1q receptor is approximately 68-kD, exhibits varying expression on different subjects, and is not upregulated from intracellular stores on exposure to soluble stimuli. Stimulated, but not resting, neutrophils selectively respond to raised levels of free C1q, resulting in altered cell function and enhanced CR3 receptor expression. These studies thus suggest complex roles for C1q in neutrophil function.
Eggleton
P
P
William B. Castle Hematology Research Laboratory, Boston City Hospital, MA.
Ghebrehiwet
B
B
Coburn
J P
JP
Sastry
K N
KN
Zaner
K S
KS
Tauber
A I
AI
D000906:Antibodies; D015153:Blotting, Western; D002352:Carrier Proteins; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D015922:Complement C1q / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D018960:Hyaluronan Receptors; D066298:In Vitro Techniques; D007700:Kinetics; D015473:Leukemia, Promyelocytic, Acute; D016177:Macrophage-1 Antigen / Q000032:analysis / Q000096:biosynthesis; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D008970:Molecular Weight; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D010972:Platelet Activating Factor / Q000494:pharmacology; D011951:Receptors, Complement / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D013481:Superoxides / Q000097:blood; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects
D000906:Antibodies; C087299:C1QBP protein, human; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D016177:Macrophage-1 Antigen; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D010972:Platelet Activating Factor; D011951:Receptors, Complement; C032402:complement 1q receptor; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015922:Complement C1q; D013755:Tetradecanoylphorbol Acetate