A receptor for phosphatidylserine-specific clearance of apoptotic cells

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Abstract

The culmination of apoptosis in vivo is phagocytosis of cellular corpses. During apoptosis, the asymmetry of plasma membrane phospholipids is lost, which exposes phosphatidylserine externally1,2,3,4. The phagocytosis of apoptotic cells can be inhibited stereospecifically by phosphatidylserine and its structural analogues, but not by other anionic phospholipids, suggesting that phosphatidylserine is specifically recognized1,5,6,7,8,9,10. Using phage display, we have cloned a gene that appears to recognize phosphatidylserine on apoptotic cells. Here we show that this gene, when transfected into B and T lymphocytes, enables them to recognize and engulf apoptotic cells in a phosphatidylserine-specific manner. Flow cytometric analysis using a monoclonal antibody suggested that the protein is expressed on the surface of macrophages, fibroblasts and epithelial cells; this antibody, like phosphatidylserine liposomes, inhibited the phagocytosis of apoptotic cells and, in macrophages, induced an anti-inflammatory state. This candidate phosphatidylserine receptor is highly homologous to genes of unknown function in Caenorhabditis elegans and Drosophila melanogaster, suggesting that phosphatidylserine recognition on apoptotic cells during their removal by phagocytes is highly conserved throughout phylogeny.

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Figure 1: Monoclonal antibody 217 identifies a surface receptor expressed at high levels on human macrophages that recognize phosphatidylserine on apoptotic cells.
Figure 2: Monoclonal 217 inhibits uptake of apoptotic Jurkat T cells by several phagocytes.
Figure 3: Comparison of the human, mouse, Drosophila and C. elegans predicted protein sequences for the candidate PS receptor.
Figure 4: Transfection of lymphocytes with the candidate PS receptor induces PS-specific binding and uptake of apoptotic cells.
Figure 5: Treatment of J774 mouse macrophages with mAb 217, PS-containing liposomes or apoptotic cells stimulates TGF-β secretion and inhibits LPS-induced TNF-α production.

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Acknowledgements

We thank K. Christensen and D. Edwards for monoclonal antibody production; L. Remigio and D. Riches for providing mouse BMDM; M. Neville and B. Groener for the gift of HC-11 cells; J. Freed for valuable critical comments and a gift of M12.C3 B cells; C. Leslie for provision of PLB 985 cells; A. Stewart for help with culture of primary lung fibroblasts; B. Schaefer for technical advice and the gift of the pUP vector; J. Westcott for provision of the cytokine assays; B. Townend for assistance with flow cytometry and cell sorting; M. Janes for assistance with fluorescence microscopy; T. Konowal for generation of the cDNA library from J774 cells; P. Taylor for assistance with bioinformatics; R. Schlegel and P. Williamson for critical comments; and D. Voelker for technical advice and constructive critical commentary.

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Correspondence to Valerie A. Fadok.

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Fadok, V., Bratton, D., Rose, D. et al. A receptor for phosphatidylserine-specific clearance of apoptotic cells . Nature 405, 85–90 (2000) doi:10.1038/35011084

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