Abstract
Anthracycline antibiotics are inducers of an immunogenic form of apoptosis that has immunostimulatory properties because of the release of damage-associated molecular patterns. To study the mechanisms used by the innate immune system to sense this immunogenic form of cell death, we established an in vivo model of cell death induced by intraperitoneal injection of doxorubicin, a prototype of anthracyclines. The acute sterile inflammation in this model is characterized by rapid influx of neutrophils and increased levels of IL-6 and monocyte chemotactic protein-1. We demonstrate that acute inflammation induced by doxorubicin is associated with apoptosis of monocytes/macrophages and that it is specific for doxorubicin, an immunogenic chemotherapeutic. Further, the inflammatory response is significantly reduced in mice deficient in myeloid differentiation primary response gene 88 (MyD88), TLR-2 or TLR-9. Importantly, a TLR-9 antagonist reduces the recruitment of neutrophils induced by doxorubicin. By contrast, the acute inflammatory response is not affected in TRIFLps2 mutant mice and in TLR-3, TLR-4 and caspase-1 knockout mice, which shows that the inflammasome does not have a major role in doxorubicin-induced acute inflammation. Our findings provide important new insights into how the innate immune system senses immunogenic apoptotic cells and clearly demonstrate that the TLR-2/TLR-9-MyD88 signaling pathways have a central role in initiating the acute inflammatory response to this immunogenic form of apoptosis.
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Abbreviations
- AnnV:
-
annexin V
- ATP:
-
adenosine triphosphate
- CDAM:
-
cell death-associated molecule
- CRT:
-
calreticulin
- DAMP:
-
damage-associated molecular pattern
- HMGB1:
-
high mobility group box 1 protein
- HSP:
-
heat shock protein
- i.p.:
-
intraperitoneal
- IL:
-
interleukin
- LPS:
-
lipopolysaccharide
- MCP-1:
-
monocyte chemotactic protein-1
- MSU:
-
monosodium urate
- MTC:
-
mitomycin c
- MYD88:
-
myeloid differentiation primary response gene 88
- NLR:
-
the nucleotide-binding oligomerization domain (NOD)-like receptor
- PAMP:
-
pathogen-associated molecular pattern
- PEC:
-
peritoneal exudate cell
- PRR:
-
pattern-recognition receptor
- RLR:
-
RIG-I-like receptor
- TGF-β:
-
transforming growth factor beta
- TLR:
-
Toll-like receptor
- TRIF:
-
Toll/IL-1R domain-containing adaptor inducing IFN-α
- UTP:
-
uridine triphosphate
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Acknowledgements
We thank Professor S Akira (for TLR-4−/− mice), Professor B Beutler (for TRIFLps2 mice), Professor G Lauvau (for Myd88−/− mice), Professor R Beyaert and J Maelfait (for TLR-3−/− mice). This work was supported by the Fund for Scientific Research Flanders (FWO-Vlaanderen, 3G072810 to D.V.K.) and by an individual research grant from FWO-Vlaanderen (31507110 to D.V.K). D.V.K is a postdoctoral fellow and AK is a doctoral fellow, both paid by fellowships from FWO-Vlaanderen. Research in the Vandenabeele unit has been supported by Flanders Institute for Biotechnology (VIB), by European grants (FP6 ApopTrain, MRTN-CT-035624; FP7 EC RTD Integrated Project, Apo-Sys, FP7-200767; Euregional PACT II), Belgian grants (Interuniversity Attraction Poles, IAP 6/18) Flemish grants (Fonds Wetenschappelijke Onderzoek Vlaanderen, G.0875.11 and G.0973.11), and Ghent University grants (MRP, GROUP-ID). PV is holder of a Methusalem grant (BOF09/01M00709) from the Flemish Government. We thank Dr. Amin Bredan (DMBR-VIB, Ghent) for editing the paper.
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Krysko, D., Kaczmarek, A., Krysko, O. et al. TLR-2 and TLR-9 are sensors of apoptosis in a mouse model of doxorubicin-induced acute inflammation. Cell Death Differ 18, 1316–1325 (2011). https://doi.org/10.1038/cdd.2011.4
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DOI: https://doi.org/10.1038/cdd.2011.4
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