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.
Subscribe to Journal
Get full journal access for 1 year
We are sorry, but there is no personal subscription option available for your country.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Fadok,V. A. et al. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148, 2207–2216 (1992).
Martin,S. J. et al. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J. Exp. Med. 182 , 1545–1556 (1995).
Verhoven,B., Schlegel,R. A. & Williamson, P. Mechanisms of phosphatidylserine exposure, a phagocyte recognition signal, on apoptotic T lymphocytes. J. Exp. Med. 182, 1597–1601 (1995).
van den Eijnde,S. M. et al. Phosphatidylserine exposure by apoptotic cells is phylogenetically conserved. Apoptosis 3, 9– 16 (1998).
Fadok,V. A. et al. Different populations of macrophages use either the vitronectin receptor or the phosphatidylserine receptor to recognize and remove apoptotic cells. J. Immunol. 149, 4029– 4035 (1992).
Fadok,V. A. et al. Particle digestibility is required for induction of the phosphatidylserine recognition mechanism used by murine macrophages to phagocytose apoptotic cells. J. Immunol. 151, 4274– 4285 (1993).
Fadok,V. A., Warner,M. L., Bratton,D. L. & Henson,P. M. CD36 is required for phagocytosis of apoptotic cells by human macrophages that use either a phosphatidylserine receptor or the vitronectin receptor (αvβ3). J. Immunol. 161, 6250 –6257 (1998).
Pradhan,D., Krahling,S., Williamson,P. & Schlegel,R. A. Multiple systems for recognition of apoptotic lymphocytes by macrophages. Mol. Biol. Cell. 8, 767– 778 (1997).
Bennett,M. R., Gibson,D. F., Schwartz,S. M. & Tait,J. F. Binding and phagocytosis of apoptotic vascular smooth muscle cells is mediated in part by exposure of phosphatidylserine. Circ. Res. 77, 1136–1142 (1995).
Shiratsuchi,A., Umeda,M., Ohba,Y. & Nakanishi,Y. Recognition of phosphatidylserine on the surface of apoptotic spermatogenic cells and subsequent phagocytosis by Sertoli cells of the rat. J. Biol. Chem. 272, 2354–2358 (1997).
Savill,J., Hogg,N., Ren,Y. & Haslett,C. Thrombospondin cooperates with CD36 and the vitronectin receptor in macrophage recognition of neutrophils undergoing apoptosis. J. Clin. Invest. 90, 1513–1522 (1992).
Sambrano,G. R. & Steinberg,D. Recognition of oxidatively damaged and apoptotic cells by an oxidized low density lipoprotein receptor on mouse peritoneal macrophages: role of membrane phosphatidylserine. Proc. Natl Acad. Sci. USA 92, 1396– 1400 (1995).
Devitt,A. et al. Human CD14 mediates recognition and phagocytosis of apoptotic cells. Nature 392, 505– 509 (1998).
Oka,K. et al. Lectin-like oxidized low-density lipoprotein receptor 1 mediates phagocytosis of aged/apoptotic cells in endothelial cells. Proc. Natl Acad. Sci. USA 95, 9535–9540 (1998).
Balasubramanian,K., Chandra,J. & Schroit, A. J. Immune clearance of phosphatidylserine-expressing cells by phagocytes. The role of β2-glycoprotein I in macrophage recognition. J. Biol. Chem. 272, 31113– 31117 (1997).
Rigotti,A., Acton,S. L. & Krieger, M. The class B scavenger receptors SR-BI and CD36 are receptors for anionic phospholipids. J. Biol. Chem. 270, 16221–16224 (1995).
Wang,P. Y., Kitchens,R. L. & Munford, R. S. Phosphatidylinositides bind to plasma membrane CD14 and can prevent monocyte activation by bacterial lipopolysaccharide. J. Biol. Chem. 273, 24309–24313 (1998).
Ryeom,S. W., Sparrow,J. R. & Silverstein, R. L. CD36 participates in the phagocytosis of rod outer segments by retinal pigment epithelium. J. Cell. Sci. 109, 387–395 (1996).
Shiratsuchi,A., Kawasaki,Y., Ikemoto,M., Arai,H. & Nakanishi,Y. Role of class B scavenger receptor type I in phagocytosis of apoptotic rat spermatogenic cells by Sertoli cells. J. Biol. Chem. 274, 5901–5908 ( 1999).
Nagase,T. et al. Prediction of the coding sequences of unidentified human genes. IX. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro. DNA Res. 5, 31–39 (1998).
Heijne,G. Membrane protein structure prediction, hydrophobicity analysis, and the positive-inside rule. J. Mol. Biol. 225, 487– 494 (1992).
Igarashi,K. et al. A novel phosphatidylserine binding peptide motif defined by an anti-idiotypic monoclonal antibody. Localization of phosphatidylserine-specific binding sites on protein kinase C and phosphatidylserine decarboxylase. J. Biol. Chem. 270, 29075–29078 (1995).
Savill,J., Dransfield,I., Hogg,N. & Haslett,C. Vitronectin receptor-mediated phagocytosis of cells undergoing apoptosis. Nature 343, 170–173 (1990).
Fadok,V. A. et al. Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF. J. Clin. Invest. 101, 890–898 (1998).
McDonald,P. P., Fadok,V. A., Bratton,D. L. & Henson,P. M. Transcriptional and translational regulation of inflammatory mediator production by endogenous TGF-β in macrophages that have ingested apoptotic cells. J. Immunol. 163, 6164– 6172 (2000)
Aramaki,Y., Matsuno,R., Nitta,F., Arima,H. & Tsuchiya,S. Negatively charged liposomes inhibit tyrosine phosphorylation of 41-kDa protein in murine macrophages stimulated with LPS. Biochem. Biophys. Res. Commun. 231, 827–830 (1997).
Wu,Y. C. & Horvitz,H. R. C. elegans phagocytosis and cell-migration protein CED-5 is similar to human DOCK180. Nature 392, 501–504 ( 1998).
Wu,Y. C. & Horvitz,H. R. The C. elegans cell corPSe engulfment gene ced-7 encodes a protein similar to ABC transporters. Cell 93, 951–960 ( 1998).
Liu,Q. A. & Hengartner,M. O. Candidate adaptor protein CED-6 promotes the engulfment of apoptotic cells in C. elegans. Cell 93, 961–972 ( 1998).
Franc,N. C., Heitzler,P., Ezekowitz,R. A. & White,K. Requirement for croquemort in phagocytosis of apoptotic cells in Drosophila . Science 284, 1991– 1994 (1999).
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.
About this article
Cite this article
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
Advanced Materials (2019)
Targeting Tyro3, Axl and MerTK (TAM receptors): implications for macrophages in the tumor microenvironment
Molecular Cancer (2019)
1,3-dimethyl-6-nitroacridine derivatives induce apoptosis in human breast cancer cells by targeting DNA
Drug Development and Industrial Pharmacy (2019)
Advanced Drug Delivery Reviews (2019)
International Immunopharmacology (2019)