Granzyme B of cytotoxic T cells induces extramitochondrial reactive oxygen species production via caspase-dependent NADPH oxidase activation
Induction of reactive oxygen species (ROS) is a hallmark of granzyme B (gzmB)-mediated pro-apoptotic processes and target cell death. However, it is unclear to what extent the generated ROS derive from mitochondrial and/or extra-mitochondrial sources. To clarify this point, we have produced a mutant...
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| Hauptverfasser: | , , , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
02 February 2010
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| In: |
Immunology & cell biology
Year: 2010, Jahrgang: 88, Heft: 5, Pages: 545-554 |
| ISSN: | 1440-1711 |
| DOI: | 10.1038/icb.2010.5 |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://dx.doi.org/10.1038/icb.2010.5 Verlag, lizenzpflichtig, Volltext: https://onlinelibrary.wiley.com/doi/epdf/10.1038/icb.2010.5 
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| Verfasserangaben: | Juan I. Aguiló, Alberto Anel, Elena Catalán, Alvaro Sebastián, Rebeca Acín-Pérez, Javier Naval, Reinhard Wallich, Markus M. Simon, Julián Pardo |
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| 245 | 1 | 0 | |a Granzyme B of cytotoxic T cells induces extramitochondrial reactive oxygen species production via caspase-dependent NADPH oxidase activation |c Juan I. Aguiló, Alberto Anel, Elena Catalán, Alvaro Sebastián, Rebeca Acín-Pérez, Javier Naval, Reinhard Wallich, Markus M. Simon, Julián Pardo |
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| 520 | |a Induction of reactive oxygen species (ROS) is a hallmark of granzyme B (gzmB)-mediated pro-apoptotic processes and target cell death. However, it is unclear to what extent the generated ROS derive from mitochondrial and/or extra-mitochondrial sources. To clarify this point, we have produced a mutant EL4 cell line, termed EL4-rho(0), which lacks mitochondrial DNA, associated with a decreased mitochondrial membrane potential and a defective ROS production through the electron transport chain of oxidative phosphorylation. When incubated with either recombinant gzmB plus streptolysin or ex vivo gzmB(+) cytotoxic T cells, EL4-rho(0) cells showed phosphatydylserine translocation, caspase 3 activation, Bak conformational change, cytochrome c release and apoptotic morphology comparable to EL4 cells. Moreover, EL4-rho(0) cells produced ROS at levels similar to EL4 under these conditions. GzmB-mediated ROS production was almost totally abolished in both cell lines by the pan-caspase inhibitor, Z-VAD-fmk. However, addition of apocynin, a specific inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, led to a significant reduction of ROS production and cell death only in EL4-rho(0) but not EL4 cells. These data suggest that gzmB-induced cell death is accompanied by a caspase-dependent pathway of extra-mitochondrial ROS production, most probably through activation of NADPH oxidase. | ||
| 650 | 4 | |a Animals | |
| 650 | 4 | |a Apoptosis | |
| 650 | 4 | |a Caspases | |
| 650 | 4 | |a Cell Line | |
| 650 | 4 | |a Enzyme Activation | |
| 650 | 4 | |a Granzymes | |
| 650 | 4 | |a Humans | |
| 650 | 4 | |a Immunoblotting | |
| 650 | 4 | |a Mice | |
| 650 | 4 | |a Mice, Inbred C57BL | |
| 650 | 4 | |a Mitochondria | |
| 650 | 4 | |a NADPH Oxidases | |
| 650 | 4 | |a Reactive Oxygen Species | |
| 650 | 4 | |a Reverse Transcriptase Polymerase Chain Reaction | |
| 650 | 4 | |a Signal Transduction | |
| 650 | 4 | |a T-Lymphocytes, Cytotoxic | |
| 700 | 1 | |a Anel, Alberto |e VerfasserIn |4 aut | |
| 700 | 1 | |a Catalán, Elena |e VerfasserIn |4 aut | |
| 700 | 1 | |a Sebastián, Alvaro |e VerfasserIn |4 aut | |
| 700 | 1 | |a Acín-Pérez, Rebeca |e VerfasserIn |4 aut | |
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| 700 | 1 | |a Simon, Markus M. |e VerfasserIn |4 aut | |
| 700 | 1 | |a Pardo, Julián |e VerfasserIn |4 aut | |
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