Noncytolytic terminal complement complexes may serve as calcium gates to elicit leukotriene B4 generation in human polymorphonuclear leukocytes.
Complement effects on human polymorphonuclear leukocytes (PMN) have generally been ascribed to the anaphylatoxin C5a, which induces degranulation, superoxide anion generation, migration, and cell aggregation via interaction with membrane receptors. We here report that complement activation on the surface of antibody-sensitized human PMN provokes generation of the potent lipid mediator leukotriene B4 (LTB4) in strict dependence on complement component C8, but in the absence of detectable C9. The kinetics of LT generation are rapid, comparable with those observed after challenge with the calcium-ionophore A23187. LTB4 release is a distinct event that is dissociable from cytotoxicity as assessed by lactate dehydrogenase (LDH) release (dependent on C9) and from superoxide generation (independent of C8 and C9). It is dose dependent on extracellular calcium and is not observed in the absence of calcium. It is inhibited by substances interfering with calcium-calmodulin function (trifluoperazine and W7), but not by blockers of physiologic calcium channels (nimodipine, verapamil, and D 888). Addition of purified C8 to cells bearing C5b-7 induces a severalfold increase in their passive permeability to 45calcium. Sieving experiments with the use of marker molecules of different sizes collectively indicate the existence of small hydrophilic channels consisting exclusively or predominantly of C5b-8 complexes, which allow passive transmembrane flux of small molecules with Mr less than 200. Thus, noncytolytic terminal complement complexes may serve as a biological bypass gate for calcium in PMN membranes, triggering the arachidonic acid cascade with generation of LTB4 at doses well below the threshold required to invoke overt cell damage.
D002118:Calcium / Q000502:physiology*; D003167:Complement Activation / Q000187:drug effects; D015938:Complement Membrane Attack Complex; D003165:Complement System Proteins / Q000502:physiology*; D003911:Dextrans / Q000378:metabolism; D004306:Dose-Response Relationship, Immunologic; D005110:Extracellular Space / Q000378:metabolism; D006801:Humans; D007473:Ion Channels / Q000502:physiology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007975:Leukotriene B4 / Q000096:biosynthesis*; D046911:Macromolecular Substances; D009504:Neutrophils / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism
D015938:Complement Membrane Attack Complex; D003911:Dextrans; D007473:Ion Channels; D046911:Macromolecular Substances; D013481:Superoxides; D007975:Leukotriene B4; D003165:Complement System Proteins; D007770:L-Lactate Dehydrogenase; D002118:Calcium