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The Journal of Immunology, Vol 144, Issue 9 3498-3505, Copyright © 1990 by American Association of Immunologists

ARTICLES

The influence of calcium, sodium, and the Na+/Ca2+ antiport on susceptibility to cytolysin/perforin-mediated cytolysis

RP Kraut, D Bose, EJ Cragoe Jr and AH Greenberg
Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Canada.

In NK cell-mediated cytolysis, the degranulation of the NK cell, and the binding and polymerization of its effector molecule cytolysin/perforin, are Ca2+ dependent. To determine if increases in the target cell free intracellular Ca2+ concentration ([Ca2+]i) are also important for cytotoxicity, we examined the effects of Ca2+ substitutes, promotors of Ca2+ influx; and inhibitors of Ca2+ efflux on the cytolysis mediated by purified cytolysin. Ca2+ promoted cytolysin- mediated cytolysis but could also inactivate cytolysin on preincubation in the absence of target cells. Ba2+ and Sr2+ could not alone promote cytolysin-mediated cytolysis but did enhance Ca2(+)-driven cytolysis. Preincubation with Ba2+ and Sr2+ did not inactivate cytolysin. One interpretation which these results suggested was that neither Ba2+ nor Sr2+ were involved in cytolysin polymerization and pore formation, but could act intracellularly as Ca2+ analogs once pore formation had occurred. The synergistic interaction of cytolysin and the Ca2+ ionophore A23187 further supported a role for increased [Ca2+]i in cytolysis. Inhibitors of Na+/Ca2+ exchange, namely low Na+ medium, ouabain, and the amiloride analogs 2',4'-dimethylbenzamil, 5-(N-4- chlorobenzyl)-2',4'dimethylbenzamil, and alpha-phenylbenzamil all markedly enhanced cytolysin-mediated cytolysis. These results support the hypothesis that the Ca2+ dependency of NK cell- and cytolysin- mediated cytolysis is related to increases in target cell [Ca2+]i. Furthermore, they indicate that Na+/Ca2+ exchange is an important counterlytic mechanism.


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M. P. Blaustein and W. J. Lederer
Sodium/Calcium Exchange: Its Physiological Implications
Physiol Rev, July 1, 1999; 79(3): 763 - 854.
[Abstract] [Full Text] [PDF]


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