title: C5a and formyl peptide receptor regulation on human monocytes.
abstract: Regulation of C5a and formyl-methionine-leucine-phenylalanine-lysine (fMLPL) receptors on human monocytes has been studied using fluorescein-conjugated derivatives and flow cytometry. Monocytes have receptors for each of these ligands, as evidenced by their ability to bind specifically biologically active fluorescein derivatives of these ligands. Quenching experiments showed that bound fluoresceinated C5a and fMLPL are rapidly internalized at 37 degrees C. Once internalized, monocytes are able to reexpress these receptors, returning to control levels within approximately 90 min. This contrasts with rate differences seen in polymorphonuclear neutrophils (PMNs), where fMLPL receptors return more rapidly (approximately 30 min) than do C5a receptors (approximately 100 min). Monensin inhibited the reexpression of C5a but not fMLPL receptors, suggesting that a receptor recycling process is necessary to replenish C5a receptors on the monocyte surface. Similar although less efficient inhibition of C5a receptor reexpression was observed with NH4Cl treatment. Reexpression of both C5a and fMLPL receptors was independent of extracellular Ca2+. Treatment with various agents known to stimulate monocytes and PMNs increased the expression of fMLPL receptors in both cell types but either had no effect on or reduced the level of C5a receptor expression. This would indicate that monocytes, like PMNs, have intracellular pools of preformed fMLPL receptors, available for reexpression. These studies show that, like PMNs, monocytes modulate C5a and fMLPL receptors through different mechanisms. Furthermore, monocytes are capable of reexpressing these receptors following exposure to ligand, a theoretical requirement for chemotaxis.
authors.affiliation: Applied Sciences Division, Baxter Healthcare Corporation, Round Lake, Illinois 60073.
authors.forename: D E
authors.initials: DE
authors.lastname: Van Epps
authors.forename: S J
authors.initials: SJ
authors.lastname: Simpson
authors.forename: D E
authors.initials: DE
authors.lastname: Chenoweth
mesh_terms: D000643:Ammonium Chloride / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D015936:Complement C5a / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D066298:In Vitro Techniques; D008985:Monensin / Q000494:pharmacology; D009000:Monocytes / Q000502:physiology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D009504:Neutrophils / Q000502:physiology; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement / Q000502:physiology*; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000502:physiology*
chemical_list: D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D000643:Ammonium Chloride; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015936:Complement C5a; D008985:Monensin; D002118:Calcium