Mitochondria-dependent and -independent regulation of granzyme B-induced apoptosis

被引:165
作者
MacDonald, G
Shi, LF
Velde, CV
Lieberman, J
Greenberg, AH [1 ]
机构
[1] Univ Manitoba, Manitoba Inst Cell Biol, Winnipeg, MB R3E 0V9, Canada
[2] Harvard Univ, Sch Med, Ctr Blood Res, Boston, MA 02115 USA
关键词
granzyme B; apoptosis; mitochondria; cytochrome c; caspase;
D O I
10.1084/jem.189.1.131
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Granzyme B (GraB) is required for the efficient activation of apoptosis by cytotoxic T lymphocytes and natural killer cells. We find that GraB and perforin induce severe mitochondrial perturbation as evidenced by the release of cytochrome c into the cytosol and suppression of transmembrane potential (Delta psi). The earliest mitochondrial event was the release of cytochrome c, which occurred at the same time as caspase 3 processing and consistently before the activation of apoptosis. Granzyme K/perforin or perforin treatment, both of which kill target cells efficiently but are poor activators of apoptosis ill short-term assays, did not induce rapid cytochrome c release. However, they suppressed Delta psi and increased reactive oxygen species generation, indicating that mitochondrial dysfunction is also associated with this nonapoptotic cell death. Pretreatment with peptide caspase inhibitors zVAD-FMK or YVAD-CHO prevented GraB apoptosis and cytochrome c release, whereas DEVD-CHO blocked apoptosis but did not prevent cytochrome c release, indicating that caspases act both up- and downstream of mitochondria. Of additional interest, Delta psi suppression mediated by GraK or GraB and perforin was not affected by zVAD-FMK and thus was caspase independent. Overexpression of Bcl-2 and Bcl-X-L suppressed caspase activation, mitochondrial cytochrome c release, Delta psi suppression, and apoptosis and cell death induced by GraB, Grak, or perforin. In an in vitro cell free system, GraB activates nuclear apoptosis in S-100 cytosol at high doses, however the addition of mitochondria amplified GraB activity over 15-fold. GraB-induced caspase 3 processing to p17 in S-100 cytosol was increased only threefold in the presence of mitochondria, suggesting that another caspase(s) participates in the: mitochondrial amplification of GraB apoptosis. We conclude that GraB-induced apoptosis is highly amplified by mitochondria in a caspase-dependent manner but that GraB can also initiate caspase 3 processing and apoptosis in the absence of mitochondria.
引用
收藏
页码:131 / 143
页数:13
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