Abstract
Phagocytosis of apoptotic cells is required to prevent tissue injury. Professional phagocytes, such as monocyte-derived macrophages, are highly efficient scavengers of apoptotic cells but their presence cannot always be relied on; in that case, removal of effete cells is accomplished by helpful neighbours. This study describes differences in the efficiency with which apoptotic cells of the same type, but dying in response to different triggers, are engulfed; this varies from engulfment that is so proficient few or no unengulfed apoptotic cells are found, to engulfment that is so delayed apoptotic cells have become secondarily necrotic at the point of engulfment. In all cases the efficiency of engulfment is determined at least in part by the dying cells themselves. p53- and Bax-transfected kidney epithelial (293) cells (transiently transfected using a non-toxic method) were engulfed so proficiently by homotypic neighbours that cells did not show evidence of engagement of the apoptotic programme (chromatin condensation and TUNEL positivity) until engulfment had taken place. Engulfment nonetheless required activation of at least initiator caspases. 293 cells induced to apoptose by other means (etoposide and staurosporine treatment) were not so efficiently ingested: unengulfed apoptotic cells were consistently revealed at all doses and time points, even when treated cells were mixed with healthy, non-treated 293 cells. These data make it extremely unlikely that the fraction of viable, unaffected neighbours determines the efficiency with which engulfment proceeds. Furthermore, 293 cells treated with etoposide or staurosporine were differentially appealing both to homotypic neighbours and to cells in the professional phagocyte lineage (THP-1 cells). If different apoptotic stimuli programme cells to be recognised with different efficiencies, pathways to apoptosis may be injury limiting to greater or lesser degrees. Cell Death and Differentiation (2001) 8, 734–746
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Abbreviations
- CMTMR:
-
5-(and 6)-(((4-chloromethyl)benzoyl)amino)tetramethylrhodamine
- CMFDA:
-
5-chloromethylfluorescein diacetate
- DIC:
-
differential interference contrast
- GFP:
-
green fluorescent protein
- PI:
-
propidium iodide
- PS:
-
phosphatidylserine
- TUNEL:
-
terminal deoxynucleotide transferase-mediated dUTP-fluorescein nick-end labelling
- ZVAD-fmk:
-
benzyloxycarbonyl-valinyl-alanyl-aspartyl-(OMe)-fluoromethyl ketone
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Acknowledgements
We would like to thank Carol Midgeley, David Lane and other members of the laboratory for generous provision of the cell line Saos2::p53teti, p53 expression vectors, and p53 antibodies. Furthermore we thank K Ryan and K Vousden for kindly providing us with pcDNA3/p53, and A Chen and CB Thompson for the generous gift of psFFVneo-Bax. We are grateful to Simon Brown and Niall McTavish for helpful discussion. This work was funded by the Wellcome Trust grant no. 033790. B Spruce is a Wellcome Senior Research Fellow in Clinical Science.
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Wiegand, U., Corbach, S., Prescott, A. et al. The trigger to cell death determines the efficiency with which dying cells are cleared by neighbours. Cell Death Differ 8, 734–746 (2001). https://doi.org/10.1038/sj.cdd.4400867
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DOI: https://doi.org/10.1038/sj.cdd.4400867
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