Skip to main content
SpringerLink
Log in

Detection and characterization of soluble CD93 released during inflammation

  • Original Research Paper
  • Published:
Inflammation Research Aims and scope Submit manuscript

Abstract

Background

CD93 is a cell surface glycoprotein that is required for efficient engulfment of apoptotic cells via an unknown mechanism. Recently, it was demonstrated that CD93 is proteolytically cleaved from the surface of activated human monocytes and neutrophils in response to inflammatory signals in vitro and that a soluble form of CD93 (sCD93) exists in human plasma.

Objective

The objective of this study was to examine the relationship among production of sCD93, inflammation, and engulfment of apoptotic cells.

Methods

Sterile peritonitis was induced in C57BL/6 mice, and lavage fluid was analyzed for presence of sCD93 by ELISA and Western blot. Cellular infiltrate was assessed by flow cytometry and analyzed for its capacity to shed CD93 in vitro. Peritoneal lavage fluid (PLF) was examined for its ability to regulate engulfment of apoptotic cells.

Results

There was an 8.9-fold increase in sCD93 following induction of peritonitis. Macrophages accounted for the majority of infiltrating leukocytes into the peritoneum when sCD93 levels were highest. Inflammatory peritoneal macrophages constitutively shed CD93 in vitro suggesting that this cell type contributes to elevated levels of sCD93 in vivo. Inflammatory PLF from wild-type mice containing elevated sCD93 significantly enhanced engulfment of apoptotic cells in vitro when compared to inflammatory fluid from CD93-deficient mice.

Conclusions

These data demonstrate that inflammation triggers release of sCD93 in vivo, identify the inflammatory macrophage as a source of sCD93, and provide insight into the mechanism by which CD93 contributes to engulfment of apoptotic cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Norsworthy PJ, Fossati-Jimack L, Cortes-Hernandez J, et al. Murine CD93 (C1qRp) contributes to the removal of apoptotic cells in vivo but is not required for C1q-mediated enhancement of phagocytosis. J Immunol. 2004;172:3406–14.

    PubMed  CAS  Google Scholar 

  2. deCathelineau AM, Henson PM. The final step in programmed cell death: phagocytes carry apoptotic cells to the grave. Essays Biochem. 2003;39:105–17.

    PubMed  CAS  Google Scholar 

  3. Stark MA, Huo Y, Burcin TL, Morris MA, Olson TS, Ley K. Phagocytosis of apoptotic neutrophils regulates granulopoiesis via IL-23 and IL-17. Immunity. 2005;22:285–94.

    Article  PubMed  CAS  Google Scholar 

  4. Erwig LP, Henson PM. Immunological consequences of apoptotic cell phagocytosis. Am J Pathol. 2007;171:2–8.

    Article  PubMed  CAS  Google Scholar 

  5. Teder P, Vandivier RW, Jiang D, et al. Resolution of lung inflammation by CD44. Science. 2002;296:155–8.

    Article  PubMed  CAS  Google Scholar 

  6. Cvetanovic M, Ucker DS. Innate immune discrimination of apoptotic cells: repression of proinflammatory macrophage transcription is coupled directly to specific recognition. J Immunol. 2004;172:880–9.

    PubMed  CAS  Google Scholar 

  7. Voll RE, Herrmann M, Roth EA, Stach C, Kalden JR, Girkontaite I. Immunosuppressive effects of apoptotic cells. Nature. 1997;390:350–1.

    Article  PubMed  CAS  Google Scholar 

  8. Fadok VA, Bratton DL, Konowal A, Freed PW, Westcott JY, Henson PM. 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. 1998;101:890–8.

    Article  PubMed  CAS  Google Scholar 

  9. Huynh ML, Fadok VA, Henson PM. Phosphatidylserine-dependent ingestion of apoptotic cells promotes TGF-beta1 secretion and the resolution of inflammation. J Clin Invest. 2002;109:41–50.

    PubMed  CAS  Google Scholar 

  10. Nepomuceno RR, Henschen-Edman AH, Burgess WH, Tenner AJ. cDNA cloning and primary structure analysis of C1qR(P), the human C1q/MBL/SPA receptor that mediates enhanced phagocytosis in vitro. Immunity. 1997;6:119–29.

    Article  PubMed  CAS  Google Scholar 

  11. Bohlson SS, Silva R, Fonseca MI, Tenner AJ. CD93 is rapidly shed from the surface of human myeloid cells and the soluble form is detected in human plasma. J Immunol. 2005;175:1239–47.

    PubMed  CAS  Google Scholar 

  12. Jutila MA, Rott L, Berg EL, Butcher EC. Function and regulation of the neutrophil MEL-14 antigen in vivo: comparison with LFA-1 and MAC-1. J Immunol. 1989;143:3318–24.

    PubMed  CAS  Google Scholar 

  13. Kishimoto TK, Jutila MA, Berg EL, Butcher EC. Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors. Science. 1989;245:1238–41.

    Article  PubMed  CAS  Google Scholar 

  14. Champagne B, Tremblay P, Cantin A, St PY. Proteolytic cleavage of ICAM-1 by human neutrophil elastase. J Immunol. 1998;161:6398–405.

    PubMed  CAS  Google Scholar 

  15. Nagano O, Murakami D, Hartmann D, et al. Cell-matrix interaction via CD44 is independently regulated by different metalloproteinases activated in response to extracellular Ca(2+) influx and PKC activation. J Cell Biol. 2004;165:893–902.

    Article  PubMed  CAS  Google Scholar 

  16. Nagano O, Saya H. Mechanism and biological significance of CD44 cleavage. Cancer Sci. 2004;95:930–5.

    Article  PubMed  CAS  Google Scholar 

  17. Black RA, Rauch CT, Kozlosky CJ, et al. A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. Nature. 1997;385:729–33.

    Article  PubMed  CAS  Google Scholar 

  18. Boehme MW, Werle E, Kommerell B, Raeth U. Serum levels of adhesion molecules and thrombomodulin as indicators of vascular injury in severe Plasmodium falciparum malaria. Clin Investig. 1994;72:598–603.

    Article  PubMed  CAS  Google Scholar 

  19. Hundhausen C, Misztela D, Berkhout TA, et al. The disintegrin-like metalloproteinase ADAM10 is involved in constitutive cleavage of CX3CL1 (fractalkine) and regulates CX3CL1-mediated cell–cell adhesion. Blood. 2003;102:1186–95.

    Article  PubMed  CAS  Google Scholar 

  20. Gough PJ, Garton KJ, Wille PT, Rychlewski M, Dempsey PJ, Raines EW. A disintegrin and metalloproteinase 10-mediated cleavage and shedding regulates the cell surface expression of CXC chemokine ligand 16. J Immunol. 2004;172:3678–85.

    PubMed  CAS  Google Scholar 

  21. Galkina E, Tanousis K, Preece G, et al. l-Selectin shedding does not regulate constitutive T cell trafficking but controls the migration pathways of antigen-activated T lymphocytes. J Exp Med. 2003;198:1323–35.

    Article  PubMed  CAS  Google Scholar 

  22. Roach T, Slater S, Koval M, et al. CD45 regulates Src family member kinase activity associated with macrophage integrin-mediated adhesion. Curr Biol. 1997;7:408–17.

    Article  PubMed  CAS  Google Scholar 

  23. Lillis AP, Greenlee MC, Mikhailenko I, et al. Murine low-density lipoprotein receptor-related protein 1 (LRP) is required for phagocytosis of targets bearing LRP ligands but is not required for C1q-triggered enhancement of phagocytosis. J Immunol. 2008;181:364–73.

    PubMed  CAS  Google Scholar 

  24. Kask L, Trouw LA, Dahlback B, Blom AM. The C4b-binding protein-protein S complex inhibits the phagocytosis of apoptotic cells. J Biol Chem. 2004;279:23869–73.

    Article  PubMed  CAS  Google Scholar 

  25. Nauta AJ, Castellano G, Xu W, et al. Opsonization with C1q and mannose-binding lectin targets apoptotic cells to dendritic cells. J Immunol. 2004;173:3044–50.

    PubMed  CAS  Google Scholar 

  26. Xu W, Roos A, Schlagwein N, Woltman AM, Daha MR, van KC. IL-10-producing macrophages preferentially clear early apoptotic cells. Blood. 2006;107:4930–7.

    Article  PubMed  CAS  Google Scholar 

  27. Park M, Tenner AJ. Cell surface expression of C1qRP/CD93 is stabilized by O-glycosylation. J Cell Physiol. 2003;196:512–22.

    Article  PubMed  CAS  Google Scholar 

  28. Guo L, Eisenman JR, Mahimkar RM, et al. A proteomic approach for the identification of cell-surface proteins shed by metalloproteases. Mol Cell Proteomics. 2002;1:30–6.

    Article  PubMed  CAS  Google Scholar 

  29. Botto M, Dell’Agnola C, Bygrave AE, et al. Homozygous C1q deficiency causes glomerulonephritis associated with multiple apoptotic bodies. Nat Genet. 1998;19:56–9.

    Article  PubMed  CAS  Google Scholar 

  30. Fonseca MI, Carpenter PM, Park M, Palmarini G, Nelson EL, Tenner AJ. C1qR(P), a myeloid cell receptor in blood, is predominantly expressed on endothelial cells in human tissue. J Leukoc Biol. 2001;70:793–800.

    PubMed  CAS  Google Scholar 

  31. Dean YD, McGreal EP, Gasque P. Endothelial cells, megakaryoblasts, platelets and alveolar epithelial cells express abundant levels of the mouse AA4 antigen, a C-type lectin-like receptor involved in homing activities and innate immune host defense. Eur J Immunol. 2001;31:1370–81.

    Article  PubMed  CAS  Google Scholar 

  32. Shi CS, Shi GY, Hsiao SM, et al. Lectin-like domain of thrombomodulin binds to its specific ligand Lewis Y antigen and neutralizes lipopolysaccharide-induced inflammatory response. Blood. 2008;112:3661–70.

    Article  PubMed  CAS  Google Scholar 

  33. Greenlee MC, Sullivan SA, Bohlson SS. CD93 and related family members: their role in innate immunity. Curr Drug Targets. 2008;9:130–8.

    Article  PubMed  CAS  Google Scholar 

  34. Ogden CA, deCathelineau A, Hoffmann PR, 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.

    Article  PubMed  CAS  Google Scholar 

  35. Stuart LM, Ezekowitz RA. Phagocytosis: elegant complexity. Immunity. 2005;22:539–50.

    Article  PubMed  CAS  Google Scholar 

  36. Chevrier S, Genton C, Kallies A, et al. CD93 is required for maintenance of antibody secretion and persistence of plasma cells in the bone marrow niche. Proc Natl Acad Sci USA. 2009;106:3895–900.

    Article  PubMed  Google Scholar 

  37. Bohlson SS, Fraser DA, Tenner AJ. Complement proteins C1q and MBL are pattern recognition molecules that signal immediate and long-term protective immune functions. Mol Immunol. 2007;44:33–43.

    Article  PubMed  CAS  Google Scholar 

  38. Babic AM. Extracorporeal photopheresis: lighting the way to immunomodulation. Am J Hematol. 2008;83:589–91.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Jessica Morris, Ana Kozmar, Michael Kuelbs, Jonah Smith, and Mark Wacker for excellent technical assistance. These studies were initiated in the laboratory of Dr. Andrea Tenner at University of California Irvine. CD93−/− mice were provided by Dr. Marina Botto at Imperial College, London. This work was supported by American Heart Association Scientist Development Grant 0630068N to S.S.B.

Author information

Authors and Affiliations

  1. Department of Biological Sciences, Eck Institute for Global Health, University of Notre Dame, Notre Dame, IN, 46556, USA

    Mallary C. Greenlee, Sarah A. Sullivan & Suzanne Slater Bohlson

  2. Department of Microbiology and Immunology, Indiana University School of Medicine-South Bend, 123 Raclin Carmichael Hall, 1234 Notre Dame Avenue, South Bend, IN, 46617, USA

    Suzanne Slater Bohlson

Authors
  1. Mallary C. Greenlee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sarah A. Sullivan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Suzanne Slater Bohlson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Suzanne Slater Bohlson.

Additional information

Responsible Editor: M. Parnham.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Greenlee, M.C., Sullivan, S.A. & Bohlson, S.S. Detection and characterization of soluble CD93 released during inflammation. Inflamm. Res. 58, 909–919 (2009). https://doi.org/10.1007/s00011-009-0064-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00011-009-0064-0

Keywords

Search

Navigation