Research Article
Impaired recognition of apoptotic neutrophils by the C1q/calreticulin and CD91 pathway in systemic lupus erythematosus
Suzanne Donnelly,
Wendy Roake, Simon Brown,
Philip Young, Haley Naik, Paul Wordsworth, David A. Isenberg, Kenneth B. M. Reid, Paul Eggleton,
Suzanne Donnelly
John Radcliffe Hospital and University of Oxford, Oxford, UK
Search for more papers by this authorSimon Brown
University of Edinburgh Medical School, Edinburgh, UK
Search for more papers by this authorCorresponding Author
Paul Eggleton
University of Oxford, Oxford, and Peninsula Medical School, Exeter, UK
Institute of Biomedical and Clinical Sciences, Peninsula Medical School, Heavitree Road, Exeter, Devon EX1 2LU, UKSearch for more papers by this authorSuzanne Donnelly,
Wendy Roake, Simon Brown,
Philip Young, Haley Naik, Paul Wordsworth, David A. Isenberg, Kenneth B. M. Reid, Paul Eggleton,
Suzanne Donnelly
John Radcliffe Hospital and University of Oxford, Oxford, UK
Search for more papers by this authorSimon Brown
University of Edinburgh Medical School, Edinburgh, UK
Search for more papers by this authorCorresponding Author
Paul Eggleton
University of Oxford, Oxford, and Peninsula Medical School, Exeter, UK
Institute of Biomedical and Clinical Sciences, Peninsula Medical School, Heavitree Road, Exeter, Devon EX1 2LU, UKSearch for more papers by this authorAbstract
Objective
A deficiency in a subcomponent of C1q can result in increased susceptibility to autoimmune diseases such as systemic lupus erythematosus (SLE). The monocyte endocytic receptor CD91 is implicated in the endocytosis of apoptotic neutrophils via interactions with C1q and calreticulin. In this clinical study, we studied the binding of C1q to leukocytes and determined whether C1q bound specifically to calreticulin and CD91 on cells undergoing apoptosis in SLE.
Methods
Proximal antibody phage display, calreticulin-transfected cells, and immunocytochemical and confocal techniques were used in a comprehensive analysis of direct binding of C1q to apoptotic neutrophils that were obtained from healthy individuals and from patients with SLE. In addition, apoptotic cellular systems were assessed in vitro.
Results
C1q appeared to colocalize to apoptotic blebs on the surface of leukocytes in association with both calreticulin and CD91, as determined by phage display and transfected cell studies. However, C1q did not bind to apoptotic cells isolated from SLE patients, despite the positivity of the cells for both calreticulin and CD91. Surface expression of calreticulin decreased on neutrophils as they aged, but increased on monocytes. In an apoptotic phagocytic assay, the addition of C1q and calreticulin significantly enhanced the phagocytosis of apoptotic cell debris by monocyte-derived cells.
Conclusion
These observations indicate that neutrophils from SLE patients have a reduced ability to be recognized and removed by the C1q/calreticulin/CD91-mediated apoptotic pathway, despite the presence of main apoptotic recognition partners. This suggests that an additional component, as yet unidentified, acts as a C1q binding partner on apoptotic cells, and this component may be lacking in cells isolated from SLE patients.
REFERENCES
- 1 Walport MJ, Davies KA, Botto M. C1q and systemic lupus erythematosus. Immunobiology 1998; 199: 265–85.
- 2 Botto M. C1q knock-out mice for the study of complement deficiency in autoimmune disease. Exp Clin Immunogenet 1998; 15: 231–4.
- 3 Trinder PK, Maeurer MJ, Stoerkel SS, Loos M. Altered (oxidized) C1q induces a rheumatoid arthritis-like destructive and chronic inflammation in joint structures in arthritis-susceptible rats. Clin Immunol Immunopathol 1997; 82: 149–56.
- 4 Petry F, Loos M. Common silent mutations in all types of hereditary complement C1q deficiencies. Immunogenetics 2005; 57: 566–71.
- 5 Racila DM, Sontheimer CJ, Sheffield A, Wisnieski JJ, Racila E, Sontheimer RD. Homozygous single nucleotide polymorphism of the complement C1QA gene is associated with decreased levels of C1q in patients with subacute cutaneous lupus erythematosus. Lupus 2003; 12: 124–32.
- 6 Dini L, Autuori F, Lentini A, Oliverio S, Piacentini M. The clearance of apoptotic cells in the liver is mediated by the asialoglycoprotein receptor. FEBS Lett 1992; 296: 174–8.
- 7 Martin SJ, Reutelingsperger CP, McGahon AJ, Rader JA, van Schie RC, LaFace DM, 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 1995; 182: 1545–56.
- 8 Savill J, Fadok V. Corpse clearance defines the meaning of cell death. Nature 2000; 407: 784–8.
- 9 Vandivier RW, Ogden CA, Fadok VA, Hoffmann PR, Brown KK, Botto M, et al. 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 2002; 169: 3978–86.
- 10
Henson PM.
A role for calreticulin in the clearance of apoptotic cells and in the innate immune system. In:
P Eggleton,
M Michalak, editors.
Calreticulin.
Georgetown:
Landes Bioscience;
2003. p.
151–61.
10.1007/978-1-4419-9258-1_15 Google Scholar
- 11 Ogden CA, deCathelineau A, Hoffmann PR, Bratton D, Ghebrehiwet B, Fadok VA, et al. C1q and mannose binding lectin engagement of cell surface calreticulin and CD91 initiates macropinocytosis and uptake of apoptotic cells. J Exp Med 2001; 194: 781–95.
- 12 Dupuis M, Schaerer E, Krause KH, Tschopp J. The calcium-binding protein calreticulin is a major constituent of lytic granules in cytolytic T lymphocytes. J Exp Med 1993; 177: 1–7.
- 13 Eggleton P, Lieu TS, Zappi EG, Sastry K, Coburn J, Zaner KS, et al. Calreticulin is released from activated neutrophils and binds to C1q and mannan-binding protein. Clin Immunol Immunopathol 1994; 72: 405–9.
- 14 Kinoshita G, Purcell AW, Keech CL, Farris AD, McCluskey J, Gordon TP. Molecular chaperones are targets of autoimmunity in Ro(SS-A) immune mice. Clin Exp Immunol 1999; 115: 268–74.
- 15 Kishore U, Sontheimer RD, Sastry KN, Zappi EG, Hughes GR, Khamashta MA, et al. The systemic lupus erythematosus (SLE) disease autoantigen-calreticulin can inhibit C1q association with immune complexes. Clin Exp Immunol 1997; 108: 181–90.
- 16 Eggleton P, Reid KB, Tenner AJ. C1q: how many functions? How many receptors? Trends Cell Biol 1998; 8: 428–31.
- 17 Kovacs H, Campbell ID, Strong P, Johnson S, Ward FJ, Reid KB, et al. Evidence that C1q binds specifically to CH2-like immunoglobulin γ motifs present in the autoantigen calreticulin and interferes with complement activation. Biochemistry 1998; 37: 17865–74.
- 18 Ronnelid J, Gunnarsson I, Nilsson-Ekdahl K, Nilsson B. Correlation between anti-C1q and immune conglutinin levels, but not between levels of antibodies to the structurally related autoantigens C1q and type II collagen in SLE or RA. J Autoimmun 1997; 10: 415–23.
- 19 Eggleton P, Ward FJ, Johnson S, Khamashta MA, Hughes GR, Hajela VA, et al. Fine specificity of autoantibodies to calreticulin: epitope mapping and characterization. Clin Exp Immunol 2000; 120: 384–91.
- 20 Van den Berg RH, Siegert CE, Faber-Krol MC, Huizinga TW, van Es LA, Daha MR. Anti-C1q receptor/calreticulin autoantibodies in patients with systemic lupus erythematosus (SLE). Clin Exp Immunol 1998; 111: 359–64.
- 21 Rosen A, Casciola-Rosen L, Ahearn J. Novel packages of viral and self-antigens are generated during apoptosis. J Exp Med 1995; 181: 1557–61.
- 22 Tan EM, Cohen AS, Fries JF, Masi AT, McShane DJ, Rothfield NF, et al. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 1982; 25: 1271–7.
- 23 Hay EM, Bacon PA, Gordon C, Isenberg DA, Maddison P, Snaith ML, et al. The BILAG index: a reliable and valid instrument for measuring clinical disease activity in systemic lupus erythematosus. Q J Med 1993; 86: 447–58.
- 24 Thiel S, Petersen SV, Vorup-Jensen T, Matsushita M, Fujita T, Stover CM, et al. Interaction of C1q and mannan-binding lectin (MBL) with C1r, C1s, MBL-associated serine proteases 1 and 2, and the MBL-associated protein MAp19. J Immunol 2000; 165: 878–87.
- 25 Goicoechea S, Pallero MA, Eggleton P, Michalak M, Murphy-Ullrich JE. The anti-adhesive activity of thrombospondin is mediated by the N-terminal domain of cell surface calreticulin. J Biol Chem 2002; 277: 37219–28.
- 26 Savill JS, Wyllie AH, Henson JE, Walport MJ, Henson PM, Haslett C. Macrophage phagocytosis of aging neutrophils in inflammation: programmed cell death in the neutrophil leads to its recognition by macrophages. J Clin Invest 1989; 83: 865–75.
- 27 Savill J, Dransfield I, Hogg N, Haslett C. Vitronectin receptor-mediated phagocytosis of cells undergoing apoptosis. Nature 1990; 343: 170–3.
- 28 McConnell JR, Crockard AD, Cairns AP, Bell AL. Neutrophils from systemic lupus erythematosus patients demonstrate increased nuclear DNA damage. Clin Exp Rheumatol 2002; 20: 653–60.
- 29 Korb LC, Ahearn JM. C1q binds directly and specifically to surface blebs of apoptotic human keratinocytes: complement deficiency and systemic lupus erythematosus revisited. J Immunol 1997; 158: 4525–8.
- 30 Navratil JS, Watkins SC, Wisnieski JJ, Ahearn JM. The globular heads of C1q specifically recognize surface blebs of apoptotic vascular endothelial cells. J Immunol 2001; 166: 3231–9.
- 31 Ghiran I, Klickstein LB, Nicholson-Weller A. Calreticulin is at the surface of circulating neutrophils and uses CD59 as an adaptor molecule. J Biol Chem 2003; 278: 21024–31.
- 32 Chan EY, Ko SC, Lau CS. Increased rate of apoptosis and decreased expression of bcl-2 protein in peripheral blood lymphocytes from patients with active systemic lupus erythematosus. Asian Pac J Allergy Immunol 1997; 15: 3–7.
- 33 Grondal G, Traustadottir KH, Kristjansdottir H, Lundberg I, Klareskog L, Erlendsson K, et al. Increased T-lymphocyte apoptosis/necrosis and IL-10 producing cells in patients and their spouses in Icelandic systemic lupus erythematosus multicase families. Lupus 2002; 11: 435–42.
- 34 Rajagopalan S, Somers EC, Brook RD, Kehrer C, Pfenninger D, Lewis E, et al. Endothelial cell apoptosis in systemic lupus erythematosus: a common pathway for abnormal vascular function and thrombosis propensity. Blood 2004; 103: 3677–83.
- 35 Pickering MC, Fischer S, Lewis MR, Walport MJ, Botto M, Cook HT. Ultraviolet-radiation-induced keratinocyte apoptosis in C1q-deficient mice. J Invest Dermatol 2001; 117: 52–8.
- 36 Eggleton P, Tenner AJ, Reid KB. C1q receptors. Clin Exp Immunol 2000; 120: 406–12.
- 37 Malhotra R, Willis AC, Jensenius JC, Jackson J, Sim RB. Structure and homology of human C1q receptor (collectin receptor). Immunology 1993; 78: 341–8.
- 38 Kumar A, Gupta R, Varghese T, Pande RM, Singal VK, Garg OP. Anti-C1q antibody as a marker of disease activity in systemic lupus erythematosus. Indian J Med Res 1999; 110: 190–3.
- 39 Racila DM, Sontheimer RD. C1q inhibits autoantibody binding to calreticulin. Lupus 1999; 8: 300–4.
- 40 Curry JL, Qin JZ, Bonish B, Carrick R, Bacon P, Panella J, et al. Innate immune-related receptors in normal and psoriatic skin. Arch Pathol Lab Med 2003; 127: 178–86.
- 41 Fabrizi C, Businaro R, Lauro GM, Fumagalli L. Role of α2-macroglobulin in regulating amyloid β-protein neurotoxicity: protective or detrimental factor? J Neurochem 2001; 78: 406–12.
- 42 Swarnakar S, Beers J, Strickland DK, Azhar S, Williams DL. The apolipoprotein E-dependent low density lipoprotein cholesteryl ester selective uptake pathway in murine adrenocortical cells involves chondroitin sulfate proteoglycans and an α2-macroglobulin receptor. J Biol Chem 2001; 276: 21121–8.
- 43 Djordjevic JT, Waterkeyn JG, Hennessy KL, Stanley KK. Role of phosphorylation of the cytoplasmic domain of the alpha(2)-macroglobulin receptor (LRP). Cell Biol Int 2000; 24: 599–610.
- 44 Taylor PR, Carugati A, Fadok VA, Cook HT, Andrews M, Carroll MC, et al. A hierarchical role for classical pathway complement proteins in the clearance of apoptotic cells in vivo. J Exp Med 2000; 192: 359–66.
- 45 Grevink ME, Horst G, Limburg PC, Kallenberg CG, Bijl M. Levels of complement in sera from inactive SLE patients, although decreased, do not influence in vitro uptake of apoptotic cells. J Autoimmun 2005; 24: 329–36.
- 46 Malhotra R, Sim RB. Chemical and hydrodynamic characterization of the human leucocyte receptor for complement subcomponent C1q. Biochem J 1989; 262: 625–31.
- 47 Erdei A, Reid KB. Characterization of C1q-binding material released from the membranes of Raji and U937 cells by limited proteolysis with trypsin. Biochem J 1988; 255: 493–9.
- 48 Eggleton P, Ghebrehiwet B, Coburn JP, Sastry KN, Zaner KS, Tauber AI. Characterization of the human neutrophil C1q receptor and functional effects of free ligand on activated neutrophils. Blood 1994; 84: 1640–9.
- 49 Ghiran I, Tyagi SR, Klickstein LB, Nicholson-Weller A. Expression and function of C1q receptors and C1q binding proteins at the cell surface. Immunobiology 2002; 205: 407–20.
- 50 Tsunoda S, Kawano M, Koni I, Kasahara Y, Yachie A, Miyawaki T, et al. Diminished expression of CD59 on activated CD8+ T cells undergoing apoptosis in systemic lupus erythematosus and Sjogren's syndrome. Scand J Immunol 2000; 51: 293–9.
- 51 Kawano M, Tsunoda S, Koni I, Mabuchi H, Muramoto H, Yachie A, et al. Decreased expression of 20-kD homologous restriction factor (HRF20, CD59) on T lymphocytes in Epstein-Barr virus (EBV)-induced infectious mononucleosis. Clin Exp Immunol 1997; 108: 260–5.
- 52 Richaud-Patin Y, Perez-Romano B, Carrillo-Maravilla E, Rodriguez AB, Simon AJ, Cabiedes J, et al. Deficiency of red cell bound CD55 and CD59 in patients with systemic lupus erythematosus. Immunol Lett 2003; 88: 95–9.
- 53 Eggleton P. Stress protein-polypeptide complexes acting as autoimmune triggers [review]. Clin Exp Immunol 2003; 134: 6–8.
