A. Sarin et al., CASPASE DEPENDENCE OF TARGET-CELL DAMAGE-INDUCED BY CYTOTOXIC LYMPHOCYTES, The Journal of immunology (1950), 161(6), 1998, pp. 2810-2816
Since the CTL secreted granule protease granzyme B can activate multip
le target caspases, it has been proposed that this pathway is responsi
ble for CTL-induced cytolysis of Fas-negative targets. However, target
lysis via the granule exocytosis pathway is completely resistant to c
aspase inhibitors. To test the possibility that granzymes trigger a po
stcaspase cytoplasmic apoptotic pathway leading to lysis, we have exam
ined the caspase dependence of several cytoplasmic changes associated
with apoptotic death. Rapid prelytic phosphatidylserine externalizatio
n was induced in Jurkat target cells by both the Fas ligand (FasL)/Fas
and the granule exocytosis effector pathways, This was specifically b
locked by peptide ketone caspase inhibitors when induced by the former
, but not by the latter, pathway. A rapid prelytic loss of target mito
chondrial psi was also induced by both CTL effector pathways, and this
was also specifically blocked by caspase inhibitors when induced by t
he FasL/Fas, but not by the granule exocytosis, pathway. Similarly, ta
rget membrane blebbing induced by CTL via the FasL/Fas, but not via th
e granule exocytosis, effector pathway was specifically blocked by cas
pase inhibitors. In contrast to the above nonnuclear damage, CTL-induc
cd target staining by the lipid probe FM1-43 reflecting plasma membran
e endocytosis was blocked by caspase inhibitors. Thus, when caspase ac
tivation is blocked, the granule exocytosis pathway triggers several p
arameters of target apoptotic damage in addition to lysis, suggesting
that granzymes directly trigger a postcaspase cytoplasmic apoptotic de
ath pathway.