Abstract
Clearance of apoptotic cells is critical to tissue homeostasis and resolution of inflammatory lesions. Macrophages are known to remove dying cells and release anti-inflammatory mediators in response; however, many cells traditionally thought of as poor phagocytes can mediate this function as well. In the lactating mammary gland following weaning, alveolar epithelial cell death is massive, yet the gland involutes rapidly, attaining its prepregnancy state in a matter of days. We found histologic evidence of apoptotic cell phagocytosis by viable mammary epithelial cells (MEC) in the involuting mouse mammary gland. Cultured MEC were able to engulf apoptotic cells in vitro, utilizing many of the same receptors used by macrophages, including the phosphatidylserine receptor (PSR), CD36, the vitronectin receptor αvβ3, and CD91. In addition, MEC, like macrophages, produced TGFβ in response to stimulation of the PSR by apoptotic cells or the anti-PSR ab 217G8E9, and downregulated endotoxin-stimulated proinflammatory cytokine production. These data support the hypothesis that amateur phagocytes play a significant role in apoptotic cell clearance and its regulation of inflammation.
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Abbreviations
- PSR:
-
phosphatidylserine receptor
- MEC:
-
mammary epithelial cell
- PMEC:
-
primary MEC
- CD91/LRP:
-
low-density lipoprotein-related receptor
- MBP:
-
mannose-binding protein
References
Geske FJ, Monks J, Lehman L and Fadok VA (2002) The role of the macrophage in apoptosis: hunter, gatherer, and regulator. Int. J. Hematol. 76: 16–26
Savill J and Fadok V (2000) Corpse clearance defines the meaning of cell death. Nature 407: 784–788
Gregory CD (2000) CD14-dependent clearance of apoptotic cells: relevance to the immune system. Curr. Opin. Immunol. 12: 27–34
Gregory CD (2000) Non-inflammatory/anti-inflammatory CD14 responses: CD14 in apoptosis. Chem. Immunol. 74: 122–140
Fadok VA, Bratton DL and Henson PM (2001) Phagocyte receptors for apoptotic cells: recognition, uptake, and consequences. J. Clin. Invest. 108: 957–962
Henson PM, Bratton DL and Fadok VA (2001) The phosphatidylserine receptor: a crucial molecular switch? Nat. Rev. Mol. Cell. Biol. 2: 627–633
Savill J, Dransfield I, Gregory C and Haslett C (2002) A blast from the past: clearance of apoptotic cells regulates immune responses. Nat. Rev. Immunol. 2: 965–975
Kreiser RJ and White K (2002) Engulfment mechanisms of apoptotic cells. Curr. Opin. Cell Biol. 14: 734–738
Balasubramanian K and Schroit AJ (2003) Aminophospholipid asymmetry: a matter of life and death. Annu. Rev. Physiol. 65: 701–734
de Freitas Balanco JM, Moreira ME, Bonoma A, Bozza PT, Amarante-Mendes G, Pirmez C and Barcinski MA (2001) Apoptotic mimicry by an obligate intracellular parasite downregulates macrophage microbicidal activity. Curr. Biol. 11: 1870–1873
Freire-de-Lima CG, Nascimento DO, Soares MB, Bozza PT, Castro-Faria-Neto HC, de Mello FG, DosReis GA and Lopes MF (2000) Uptake of apoptotic cells drives the growth of a pathogenic trypanosome in macrophages. Nature 403: 199–203
Tripathi A and Gupta CM (2003) Transbilayer translocation of membrane phosphatidylserine and its role in macrophage invasion in Leishmania promastigotes. Mol. Biochem. Parasitol. 128: 1–9
Wood W, Turmaine M, Weber R, Camp V, Maki RA, McKercher SR and Martin P (2000) Mesenchymal cells engulf and clear apoptotic footplate cells in macrophageless PU.1 null mouse embryos. Development 127: 5245–5252
Walker NI, Bennet RE and Kerr JFR (1989) Cell death by apoptosis during involution of the lactating breast in mice and rats. Am. J. Anat. 185: 19–32
Sexton DW, Blaylock MG and Walsh GM (2001) Human alveolar epithelial cells engulf apoptotic eosinophils by means of integrin- and phosphatidylserine receptor-dependent mechanisms: a process upregulated by dexamethasone. J. Allergy Clin. Immunol. 108: 962–969
Walsh GM, Sexton DW, Blaylock MG and Convery CM (1999) Resting and cytokine-stimulated human small airway epithelial cells recognize and engulf apoptotic eosinophils. Blood 94: 2827–2835
Hughes J, Liu Y, Van Damme J and Savill J (1997) Human glomerular mesangial cell phagocytosis of apoptotic neutrophils: mediation by a novel CD36-independent vitronectin receptor/thrombospondin recognition mechanism that is uncoupled from chemokine secretion. J. Immunol. 158: 4389–4397
Liu Y, Cousin JM, Hughes J, Van Damme J, Seckl JR, Haslett C, Dransfield I, Savill J and Rossi AG (1999) Glucocorticoids promote nonphlogistic phagocytosis of apoptotic leukocytes. J. Immunol. 162: 3639–3646
Savill J, Smith J, Sarraf C, Ren Y, Abbott F and Rees A (1992) Glomerular mesangial cells and inflammatory macrophages ingest neutrophils undergoing apoptosis. Kidney Int. 42: 924–936
Bennett MR, Gibson DF, Schwartz SM and Tait JF (1995) Binding and phagocytosis of apoptotic vascular smooth muscle cells is mediated in part by exposure of phosphatidylserine. Circ. Res. 77: 1136–1142
Dini L (1998) Endothelial liver cell recognition of apoptotic peripheral blood lymphocytes. Biochem. Soc. Trans. 26: 635–639
Dini L (2000) Recognizing death: liver phagocytosis of apoptotic cells. Eur. J. Histochem. 44: 217–227
Dini L, Autuori F, Lentini A, Oliverio S and Piacentini M (1992) The clearance of apoptotic cells in the liver is mediated by the asialoglycoprotein receptor. FEBS Lett. 296: 174–178
Dini L and Carla EC (1998) Hepatic sinusoidal endothelium heterogeneity with respect to the recognition of apoptotic cells. Exp. Cell Res. 240: 388–393
Dini L, Lentini A, Diez GD, Rocha M, Falasca L, Serafino L and Vidal-Vanaclocha F (1995) Phagocytosis of apoptotic bodies by liver endothelial cells. J. Cell Sci. 108: 967–973
Falasca L, Bergamini A, Serafino A, Balabaud C and Dini L (1996) Human Kupffer cell recognition and phagocytosis of apoptotic peripheral blood lymphocytes. Exp. Cell Res. 224: 152–162
Oka K, Sawamura T, Kikuta K, Itokawa S, Kume N, Kita T and Masaki T (1998) 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
Hess KL, Tudor KS, Johnson JD, Osati-Ashtiani F, Askew DS and Cook-Mills JM (1997) Human and murine high endothelial venule cells phagocytose apoptotic leukocytes. Exp. Cell Res. 236: 404–411
Boudjellab N, Chan-Tang HS, Li X and Zhao X (1998) Interleukin 8 reponse by bovine mammary epithelial cells to lipopolysaccharide stimulation. Am. J. Vet. Res. 59: 1563–1567
Boudjellab N, Chan-Tang HS and Zhao X (2000) Bovine interleukin-1 expression by cultured mammary epithelial cells (MAC-T) and its involvement in the release of MAC-T derived interleukin-8. Comp. Biochem. Physiol. A Mol. Integr. Physiol. 127: 191–199
Doppler W, Groner B and Ball RK (1989) Prolactin and glucocorticoid hormones synergistically induce expression of transfected rat beta-casein gene promoter constructs in a mammary epithelial cell line. Proc. Natl. Acad. Sci. USA 86: 104–108
Reichmann E, Ball R, Groner B and Friis RR (1989) New mammary epithelial and fibroblastic cell clones in coculture form structures competent to differentiate functionally. J. Cell Biol. 108: 1127–1138
Hoffmann PR, deCathelineau AM, Ogden CA, Leverrier Y, Bratton DL, Daleke DL, Ridley AJ, Fadok VA and Henson PM (2001) Phosphatidylserine (PS) induces PS receptor-mediated macropinocytosis and promotes clearance of apoptotic cells. J. Cell Biol. 155: 649–660
Savill J, Dransfield I, Hogg N and Haslett C (1990) Vitronectin receptor-mediated phagocytosis of cells undergoing apoptosis. Nature 343: 170–173
Savill J, Hogg N, Ren Y and Haslett C (1992) Thrombospondin cooperates with CD36 and the vitronectin receptor in macrophage recognition of neutrophils undergoing apoptosis. J. Clin. Invest. 90: 1513–1522
Ogden CA, de Cathelineau A, Hoffmann PR, Fadok VA, Bratton DL and Henson PM (2001) C1q and mannose binding lectin engagement of cell surface calreticulin and CD91 initiates macropinocytosis and uptake of apoptotic cells. J. Exp. Med. 194: 781–795
Vandivier RW, Ogden CA, Fadok VA, Hoffmann PR, Brown KK, Botto M, Walport MJ, Fisher JH, Henson PM and Greene KE (2002) Role of surfactant proteins A, D, and C1q in the clearance of apoptotic cells in vivo and in vitro: calreticulin and CD91 as a common collectin receptor complex. J. Immunol. 169: 3978–3986
Fadok VA, Bratton DL, Rose DM, Pearson A, Ezekewitz RA and Henson PM (2000) A receptor for phosphatidylserine-specific clearance of apoptotic cells. Nature 405: 85–90
Platt N, Suzuki H, Kurihara Y, Kodama T and Gordon S (1996) Role for the class A macrophage scavenger receptor in the phagocytosis of apoptotic thymocytes in vitro. Proc. Natl. Acad. Sci. USA 93: 12456–12460
Terpstra V, Kondratenko N and Steinberg D (1997) Macrophages lacking scavenger receptor A show a decrease in binding and uptake of acetylated low-density lipoprotein and of apoptotic thymocytes, but not of oxidatively damaged red blood cells. Proc. Natl. Acad. Sci. USA 94: 8127–8131
Devitt A, Moffatt OD, Raykundalia C, Capra JD, Simmons DL and Gregory CD (1998) Human CD14 mediates recognition and phagocytosis of apoptotic cells. Nature 392: 505–509
Takizawa F, Shoutaro T and Nagasawa S (1996) Enhancement of macrophage phagocytosis upon iC3b deposition on apoptotic cells. FEBS Lett. 397: 269–272
Mevorach D, Mascarenhas JO, Gershov D and Elkon KB (1998) Complement-dependent clearance of apoptotic cells by human macrophages. J. Exp. Med. 188: 2313–2320
Fadok VA, Bratton DL, Guthrie L and Henson PM (2001) Differential effects of apoptotic vs lysed cells on macrophage production of cytokines: role of proteases. J. Immunol. 161: 5873–5879
Fadok VA, Bratton DL, Konowal A, Freed PW, Westcott JY and Henson PM (1998) 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
Huynh M-L, Fadok VA and Henson PM (2002) TGFb1 secretion and resolution of inflammation are increased after phosphatidylserine-dependent ingestion of apoptotic cells. J. Clin. Invest. 109: 41–50
Voll RE, Herrmann M, Roth EA, Stach C, Kalden JR and Girkontaite I (1997) Immunosuppressive effects of apoptotic cells (letter). Nature 390: 350–351
Fadok VA, de Cathelineau A, Daleke DL, Henson PM and Bratton DL (2001) Loss of phospholipid asymmetry and surface exposure of phosphatidylserine is required for phagocytosis of apoptotic cells by macrophages and fibroblasts. J. Biol. Chem. 276: 1071–1077
Parnaik RR, Raff MC and Scholes J (2000) Differences between the clearance of apoptotic cells by professional and non-professional phagocytes. Curr. Biol. 10: 857–860
Ellis RE, Jacobson DM and Horvitz HR (1991) Genes required for the engulfment of cell corpses during programmed cell death in Caenorhabditis elegans. Genetics 129: 79–94
Su HP, Brugnera E, Van Criekinge W, Smits E, Hengartner M, Bogaert T and Ravichandran KS (2000) Identification and characterization of a dimerization domain in CED-6, an adapter protein involved in engulfment of apoptotic cells. J. Biol. Chem. 275: 9542–9549
Su HP, Nakada-Tsukui K, Tosello-Trampont AC, Li Y, Bu G, Henson PM and Ravichandran KS (2002) Interaction of CED-6/GULP, an adapter protein involved in engulfment of apoptotic cells with CED-1 and CD91/low density lipoprotein receptor-related protein (LRP). J. Biol. Chem. 277: 11772–11779
Tosello-Trampont AC, Brugnera E and Ravichandran KS (2001) Evidence for a conserved role for CRKII and Rac in engulfment of apoptotic cells. J. Biol. Chem. 276: 13797–13802
Brugnera E, Haney L, Grimsley C, Lu M, Walk SF, Tosello-Trampont AC, Macara IG, Madhani H, Fink GR and Ravichandran KS (2002) Unconventional Rac-GEF activity is mediated through the Dock180-ELMO complex. Nat. Cell Biol. 4: 574–582
Gumienny TL, Brugnera E, Tosello-Trampont AC, Kinchen JM, Haney LB, Nishiwaki K, Walk SF, Nemergut ME, Macara IG, Francis R, Schedl T, Qin Y, Van Aelst L, Hengartner MO and Ravichandran KS (2001) CED-12/ELMO, a novel member of the CrkII/Dock180/Rac pathway, is required for phagocytosis and cell migration. Cell 107: 27–41
Wang X, Wu YC, Fadok VA, Lee MC, Gengyo-Ando K, Cheng LC, Ledwich D, Hsu PK, Chen JY, Chou BK, Henson P, Mitani S and Xue D (2003) Cell corpse engulfment mediated by C. elegans phosphatidylserine receptor through CED-5 and CED-12. Science 302: 1563–1566
Ball RK, Friis RR, Schoenenberger CA, Doppler W and Groner B (1988) Prolactin regulation of beta-casein gene expression and of a cytosolic 120-kd protein in a cloned mouse mammary epithelial cell line. EMBO J. 7: 2089–2095
Emerman JT and Pitelka DR (1977) Maintenance and induction of morphological differentiation in dissociated mammary epithelium on floating collagen membranes. In vitro 13: 316–328
Merlo GR, Graus-Porta D, Cella N, Marte BM, Taverna D and Hynes NE (1996) Growth, differentiation and survival of HC11 mammary epithelial cells: diverse effects of receptor tyrosine kinase-activating peptide growth factors. Eur. J. Cell Biol. 70: 97–105
Fadok VA, Savill JS, Haslett C, Bratton DL, Doherty DE, Campbell PA and Henson PM (1992) Different populations of macrophages use either the vitronectin receptor or the phosphatidylserine receptor to recognize and remove apoptotic cells. J. Immunol. 149: 4029–4035
Kench JA, Russell DM, Fadok VA, Young SK, Worthen GS, Jones-Carson J, Henson JE, Henson PM and Nemazee D (1999) Aberrant wound healing and TGF-beta production in the autoimmune-prone MRL/+ mouse. Clin. Immunol. 92: 300–310
Acknowledgements
We thank Jay Westcott for invaluable assistance with the cytokine ELISA assays, Pat Foley at Cascade Biosciences for the gift of anti-mouse CD36 antibody, and Gail Smith for electron microscopy assistance. This work was supported by NIH grant numbers PO1 HD 38129, RO1 GM60449, and P30 CA 46934 as well as the Florence Myers Goldhamer Fellowship in Pediatric Allergy and Immunology.
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Monks, J., Rosner, D., Jon Geske, F. et al. Epithelial cells as phagocytes: apoptotic epithelial cells are engulfed by mammary alveolar epithelial cells and repress inflammatory mediator release. Cell Death Differ 12, 107–114 (2005). https://doi.org/10.1038/sj.cdd.4401517
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DOI: https://doi.org/10.1038/sj.cdd.4401517
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