The Journal of Immunology, Vol 141, Issue 4 1288-1294, Copyright © 1988 by American Association of Immunologists
Transmembrane chloride flux is required for target cell lysis but not for Golgi reorientation in cloned cytolytic effector cells. Golgi reorientation, N alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester serine esterase release, and delivery of the lethal hit are separable events in target cell lysis
G Prochazka, C Landon and G Dennert
Immunology Program, University of Southern California Comprehensive Cancer Center, Los Angeles 90033.
Cell-mediated cytotoxicity can be inhibited by the replacement of chloride
with ions that are incapable of passing through chloride channels or by the
presence of stilbene disulfonate derivatives known to interfere with
chloride flux. We show that the stilbene disulfonate
(4,4-diisothiocyano-2,2'-disulfonic acid stilbene (DIDS) inhibits lysis of
YAC-1 targets by the cloned cell line NKB61A2. Inhibition of lysis occurs
on the level of the effector cell inasmuch as preincubation of effectors
but not of targets interferes with subsequent lysis. Moreover, inhibition
of chloride flux in the target does not interfere with target cell lysis by
cytotoxic granules isolated from killer cells. Target cell binding takes
place in the presence of DIDS or absence of external chloride, suggesting
that events that follow target cell binding require chloride flux. We show
that reorientation of the Golgi apparatus, which occurs subsequent to
target cell binding in the effector cell, occurs under conditions that
interfere with chloride flux. It is therefore suggested that events in the
effector cell taking place subsequent to the Golgi apparatus reorientation
reaction are inhibited and that delivery of the lethal hit is a
stimulus-induced secretory event that requires transmembrane chloride flux.
Delivery of the lethal hit is shown to be independent of the release of N
alpha- benzyloxycarbonyl-L-lysine thiobenzyl ester (BLT) serine esterase,
suggesting that cytolytic components and BLT serine esterase are likely
packaged in different vesicles.
