Cytotoxic cells kill intracellular bacteria through granulysin-mediated delivery of granzymes

Michael Walch; Farokh Dotiwala; Sachin Mulik; Jerome Thiery; Tomas Kirchhausen; Carol Clayberger; Alan M. Krensky; Denis Martinvalet; Judy Lieberman

doi:10.1016/j.cell.2014.03.062

Cytotoxic cells kill intracellular bacteria through granulysin-mediated delivery of granzymes

Michael Walch^*, Farokh Dotiwala, Sachin Mulik, Jerome Thiery, Tomas Kirchhausen, Carol Clayberger, Alan M. Krensky, Denis Martinvalet, Judy Lieberman

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

162 Scopus citations

Abstract

When killer lymphocytes recognize infected cells, perforin delivers cytotoxic proteases (granzymes) into the target cell to trigger apoptosis. What happens to intracellular bacteria during this process is unclear. Human, but not rodent, cytotoxic granules also contain granulysin, an antimicrobial peptide. Here, we show that granulysin delivers granzymes into bacteria to kill diverse bacterial strains. In Escherichia coli, granzymes cleave electron transport chain complex I and oxidative stress defense proteins, generating reactive oxygen species (ROS) that rapidly kill bacteria. ROS scavengers and bacterial antioxidant protein overexpression inhibit bacterial death. Bacteria overexpressing a GzmB-uncleavable mutant of the complex I subunit nuoF or strains that lack complex I still die, but more slowly, suggesting that granzymes disrupt multiple vital bacterial pathways. Mice expressing transgenic granulysin are better able to clear Listeria monocytogenes. Thus killer cells play an unexpected role in bacterial defense.

Original language	English (US)
Pages (from-to)	1309-1323
Number of pages	15
Journal	Cell
Volume	157
Issue number	6
DOIs	https://doi.org/10.1016/j.cell.2014.03.062
State	Published - Jun 5 2014

Funding

This work was supported by NIH AI-045587 (to J.L.), the Stiefel-Zangger and the Kurt and Senta Herrmann Foundations (to M.W.), and PCMM-Glaxo Smith Kline Alliance (to F.D.), and by NIH grant GM-075252 (to T.K.). We thank Zhan Xu and Solange Kharoubi Hess for technical support, Luis Filgueira for helpful discussions, Marshall Thomas for HeLa-BCL2 cells, James Imlay (University of Illinois) for valuable suggestions and for the L106 E. coli strain, Susan Lovett (Brandeis) for the recA strain, Thorsten Friedrich (Albert-Ludwigs-University, Freiburg, Germany) for E. coli strain ANN0221/pBADnuo/His-nuoF, and Darren Higgins (Harvard Medical School) for the Lm expression plasmid pLIV1 and Eric Marino for maintaining the Imaging Resource used in this study.

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/j.cell.2014.03.062

Cite this

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title = "Cytotoxic cells kill intracellular bacteria through granulysin-mediated delivery of granzymes",

abstract = "When killer lymphocytes recognize infected cells, perforin delivers cytotoxic proteases (granzymes) into the target cell to trigger apoptosis. What happens to intracellular bacteria during this process is unclear. Human, but not rodent, cytotoxic granules also contain granulysin, an antimicrobial peptide. Here, we show that granulysin delivers granzymes into bacteria to kill diverse bacterial strains. In Escherichia coli, granzymes cleave electron transport chain complex I and oxidative stress defense proteins, generating reactive oxygen species (ROS) that rapidly kill bacteria. ROS scavengers and bacterial antioxidant protein overexpression inhibit bacterial death. Bacteria overexpressing a GzmB-uncleavable mutant of the complex I subunit nuoF or strains that lack complex I still die, but more slowly, suggesting that granzymes disrupt multiple vital bacterial pathways. Mice expressing transgenic granulysin are better able to clear Listeria monocytogenes. Thus killer cells play an unexpected role in bacterial defense.",

author = "Michael Walch and Farokh Dotiwala and Sachin Mulik and Jerome Thiery and Tomas Kirchhausen and Carol Clayberger and Krensky, {Alan M.} and Denis Martinvalet and Judy Lieberman",

note = "Funding Information: This work was supported by NIH AI-045587 (to J.L.), the Stiefel-Zangger and the Kurt and Senta Herrmann Foundations (to M.W.), and PCMM-Glaxo Smith Kline Alliance (to F.D.), and by NIH grant GM-075252 (to T.K.). We thank Zhan Xu and Solange Kharoubi Hess for technical support, Luis Filgueira for helpful discussions, Marshall Thomas for HeLa-BCL2 cells, James Imlay (University of Illinois) for valuable suggestions and for the L106 E. coli strain, Susan Lovett (Brandeis) for the recA strain, Thorsten Friedrich (Albert-Ludwigs-University, Freiburg, Germany) for E. coli strain ANN0221 /pBADnuo/His-nuoF , and Darren Higgins (Harvard Medical School) for the Lm expression plasmid pLIV1 and Eric Marino for maintaining the Imaging Resource used in this study. ",

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AU - Clayberger, Carol

AU - Krensky, Alan M.

AU - Martinvalet, Denis

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N2 - When killer lymphocytes recognize infected cells, perforin delivers cytotoxic proteases (granzymes) into the target cell to trigger apoptosis. What happens to intracellular bacteria during this process is unclear. Human, but not rodent, cytotoxic granules also contain granulysin, an antimicrobial peptide. Here, we show that granulysin delivers granzymes into bacteria to kill diverse bacterial strains. In Escherichia coli, granzymes cleave electron transport chain complex I and oxidative stress defense proteins, generating reactive oxygen species (ROS) that rapidly kill bacteria. ROS scavengers and bacterial antioxidant protein overexpression inhibit bacterial death. Bacteria overexpressing a GzmB-uncleavable mutant of the complex I subunit nuoF or strains that lack complex I still die, but more slowly, suggesting that granzymes disrupt multiple vital bacterial pathways. Mice expressing transgenic granulysin are better able to clear Listeria monocytogenes. Thus killer cells play an unexpected role in bacterial defense.

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Cytotoxic cells kill intracellular bacteria through granulysin-mediated delivery of granzymes

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