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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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Main Authors: Aguiló, Juan (Author) , Anel, Alberto (Author) , Catalán, Elena (Author) , Sebastián, Alvaro (Author) , Acín-Pérez, Rebeca (Author) , Naval, Javier (Author) , Wallich, Reinhard (Author) , Simon, Markus M. (Author) , Pardo, Julián (Author)
Format: Article (Journal)
Language:English
Published: 02 February 2010
In: Immunology & cell biology
Year: 2010, Volume: 88, Issue: 5, Pages: 545-554
ISSN:1440-1711
DOI:10.1038/icb.2010.5
Online Access: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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Author Notes: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. 
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650 4 |a Humans 
650 4 |a Immunoblotting 
650 4 |a Mice 
650 4 |a Mice, Inbred C57BL 
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650 4 |a NADPH Oxidases 
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650 4 |a Reverse Transcriptase Polymerase Chain Reaction 
650 4 |a Signal Transduction 
650 4 |a T-Lymphocytes, Cytotoxic 
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700 1 |a Pardo, Julián  |e VerfasserIn  |4 aut 
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