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Cited by 30Phagocytosis and Immune RegulationCoagulation, Bradykinin, Polyphosphates, and AngioedemaBlood Coagulation and Thrombosis Mechanisms

High Molecular Weight Kininogen Binds Phosphatidylserine and Opsonizes Urokinase Plasminogen Activator Receptor–Mediated Efferocytosis

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Yang, Aizhen, Dai, Jihong, Xie, Zhanli, Colman, Robert W, Wu, Qingyu, Birge, Raymond B, Wu, Yi·2014·The Journal of Immunology
DOI 10.4049/jimmunol.1302590PMC PMC4134950
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Abstract

Phagocytosis of apoptotic cells (efferocytosis) is essential for regulation of immune responses and tissue homeostasis and is mediated by phagocytic receptors. In this study, we found that urokinase plasminogen activator receptor (uPAR) plays an important role in internalization of apoptotic cells and also characterized the underlying mechanisms. In a flow cytometry–based phagocytic assay, uPAR-deficient macrophages displayed significant defect in internalization but not tethering of apoptotic cells. When uPAR-deficient mice were challenged with apoptotic cells, they exhibited pronounced splenomegaly resulting from accumulation of abundant apoptotic cells in spleen. Overexpression of uPAR in HEK-293 cells enhanced efferocytosis, which was inhibited by Annexin V and phosphatidylserine (PS) liposome, suggesting that uPAR-mediated efferocytosis is dependent on PS. In serum lacking high m.w. kininogen (HK), a uPAR ligand, uPAR-mediated efferocytosis was significantly attenuated, which was rescued by replenishment of HK. As detected by flow cytometry, HK selectively bound to apoptotic cells, but not viable cells. In purified systems, HK was specifically associated with PS liposome. HK binding to apoptotic cells induced its rapid cleavage to the two-chain form of HK (HKa) and bradykinin. Both the H chain and L chain of HKa were associated with PS liposome and apoptotic cells. HKa has higher binding affinity than HK to uPAR. Overexpression of Rac1/N17 cDNA inhibited uPAR-mediated efferocytosis. HK plus PS liposome stimulated a complex formation of CrkII with p130Cas and Dock-180 and Rac1 activation in uPAR-293 cells, but not in control HEK-293 cells. Thus, uPAR mediates efferocytosis through HK interaction with PS on apoptotic cells and activation of the Rac1 pathway.

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Authors
Yang, A., Dai, J., Xie, Z., Colman, R. W., Wu, Q., Birge, R. B., & Wu, Y.
Affiliations
  • Soochow University (CN)
  • Temple University (US)
  • Rutgers, The State University of New Jersey (US)
Corresponding author
  • Yang, Aizhen
  • Dai, Jihong
  • Xie, Zhanli
  • Wu, Qingyu
  • Wu, Yi
ORCID
Venue
The Journal of Immunology
Publisher
Oxford University Press (OUP)
Published
2014-05-01
Volume
192
Issue
9
Pages
4398-4408
ISSN
  • 0022-1767 (ISSN-L)
  • 1550-6606
DOI
10.4049/jimmunol.1302590
PMC
PMC4134950
OpenAlex
W2098224485
Cited by
30
References
1
Citations by year
2026: 2 · 2025: 6 · 2024: 1 · 2023: 1 · 2022: 4 · 2021: 1 · 2020: 2 · 2019: 1 · 2018: 2 · 2017: 3 · 2016: 3 · 2015: 3 · 2014: 1
FWCI
1.52
Type
journal article
Language
en
Open access
Licence
academic.oup.com/pages/standard-publication-reuse-rights (vor)
Locations
  • The Journal of Immunology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • Rutgers University Community Repository (Rutgers University) (submittedVersion)
Funders
  • National Natural Science Foundation of China (30971491, 81270592)
  • Priority Academic Program Development of Jiangsu Higher Education Institutions
  • National Institutes of Health
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (R03AR063290, R03AR057542)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Phagocytosis and Immune Regulation
  • Health Sciences › Medicine › Genetics › Coagulation, Bradykinin, Polyphosphates, and Angioedema
  • Health Sciences › Medicine › Hematology › Blood Coagulation and Thrombosis Mechanisms
Keywords
high molecular weight kininogen, efferocytosis, urokinase-type plasminogen activator receptor, phosphatidylserine, Rac1 signaling, CrkII, p130Cas, bradykinin, macrophages, splenomegaly, opsonization
MeSH terms
Animals, Humans, Macrophages, Mice, Inbred C57BL, Phagocytosis, Phosphatidylserines, Transfection, Immunoblotting, Signal Transduction, Apoptosis, Mice, Knockout, Kininogen, High-Molecular-Weight, Surface Plasmon Resonance, rac1 GTP-Binding Protein, Immunoprecipitation, Mice, Receptors, Urokinase Plasminogen Activator, HEK293 Cells

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