STIMULATION OF CA2+-DEPENDENT CHEMI-LUMINESCENCE IN RAT POLYMORPHONUCLEAR LEUKOCYTES BY POLYSTYRENE BEADS AND THE NONLYTIC ACTION OF COMPLEMENT

  • 1 January 1981
    • journal article
    • research article
    • Vol. 44  (3) , 569-576
Abstract
Chemiluminescence of rat polymorphonuclear leukocytes was stimulated by a phagocytic stimulus, latex beads (diameter = 1.01 .mu.m). Maximum chemiluminescent intensity increased with bead concentration in the range 0.2-20 .times. 109 beads/ml. This response was abolished in the absence of extracellular Ca2+ (1 mM EGTA [ethylene glycol bis(.beta.-aminoethyl ether) N,N,N''N''-tetraacetic acid]). Chemiluminescence could also be stimulated by the Ca2+ ionophore A23187 [calcimycin] in the presence of extracellular Ca. Addition of human serum, as a source of complement, to rat polymorphonuclear leukocytes preincubated with anti-5''-nucleotidase serum resulted in a rapid stimulation of chemiluminescence, after a lag of .apprx. 40 s. Stimulation of chemiluminescence by antibody plus complement was not the result of cell lysis because: no significant release of lactate dehydrogenase was detected at the time of the chemiluminescent response; chemiluminescence was associated with the cells and not the surrounding media; and cell lysis did not produce chemiluminescence. Chemiluminescence stimulated by antibody plus complement or by beads was inhibited by the calmodulin-blocker, trifluoperazine (50% inhibition with .apprx. 20-30 .mu.M). Cu2+ (10-4 M), which can inhibit C9 [complement component 9] action, inhibited the rapid rise in chemiluminescence induced by antibody plus complement, but not the bead-induced chemiluminescence. Depletion of C9 from human serum markedly inhibited the complement induced chemiluminescence response. Addition of purified C9 restored the response. Formation of the terminal complement attack complex at the surface of rat polymorphonuclear leukocytes apparently induces Ca2+-dependent chemiluminescence in the cells, in the absence of cell lysis.