Leukocyte integrin Mac-1 (CD11b/CD18,M2, CR3) acts as a functional receptor for platelet factor 4

Valeryi K. Lishko, Valentin P. Yakubenko, Tatiana Ugarova, Nataly Podolnikova

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Platelet factor 4 (PF4) is one of the most abundant cationic proteins secreted from -granules of activated platelets. Based on its structure, PF4 was assigned to the CXC family of chemokines and has been shown to have numerous effects on myeloid leukocytes. However, the receptor for PF4 remains unknown. Here, we demonstrate that PF4 induces leukocyte responses through the integrin Mac-1 (M2, CD11b/CD18). Human neutrophils, monocytes, U937 monocytic and HEK293 cells expressing Mac-1 strongly adhered to immobilized PF4 in a concentration-dependent manner. The cell adhesion was partially blocked by anti-Mac-1 mAb and inhibition was enhanced when anti-Mac-1 antibodies were combined with glycosaminoglycans, suggesting that cell-surface proteoglycans act cooperatively with Mac-1. PF4 also induced Mac-1-dependent migration of human neutrophils and murine WT, but not Mac-1-deficient macrophages. Coating of Escherichia coli bacteria or latex beads with PF4 enhanced their phagocytosis by macrophages by 4-fold, and this process was blocked by different Mac-1 antagonists. Furthermore, PF4 potentiated phagocytosis by WT, but not Mac-1-deficient macrophages. As determined by biolayer interferometry, PF4 directly bound the MI-domain, the major ligand-binding region of Mac-1, and this interaction was governed by a Kd of 1.3 0.2 M. Using the PF4-derived peptide library, synthetic peptides duplicating the MI-domain recognition sequences and recombinant mutant PF4 fragments, the binding sites for MI-domain were identified in the PF4 segments Cys12–Ser26 and Ala57–Ser70. These results identify PF4 as a ligand for the integrin Mac-1 and suggest that many immune-modulating effects previously ascribed to PF4 are mediated through its interaction with Mac-1.

Original languageEnglish (US)
Pages (from-to)6869-6882
Number of pages14
JournalJournal of Biological Chemistry
Volume293
Issue number18
DOIs
StatePublished - Jan 1 2018

Fingerprint

Platelet Factor 4
Integrins
Leukocytes
Macrophages
Peptide Library
Phagocytosis
Neutrophils
Interferometry
Ligands
CXC Chemokines
U937 Cells
HEK293 Cells
Cell adhesion
Latex
Proteoglycans
Platelets
Glycosaminoglycans
Microspheres
Chemokines
Cell Adhesion

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Leukocyte integrin Mac-1 (CD11b/CD18,M2, CR3) acts as a functional receptor for platelet factor 4. / Lishko, Valeryi K.; Yakubenko, Valentin P.; Ugarova, Tatiana; Podolnikova, Nataly.

In: Journal of Biological Chemistry, Vol. 293, No. 18, 01.01.2018, p. 6869-6882.

Research output: Contribution to journalArticle

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abstract = "Platelet factor 4 (PF4) is one of the most abundant cationic proteins secreted from -granules of activated platelets. Based on its structure, PF4 was assigned to the CXC family of chemokines and has been shown to have numerous effects on myeloid leukocytes. However, the receptor for PF4 remains unknown. Here, we demonstrate that PF4 induces leukocyte responses through the integrin Mac-1 (M2, CD11b/CD18). Human neutrophils, monocytes, U937 monocytic and HEK293 cells expressing Mac-1 strongly adhered to immobilized PF4 in a concentration-dependent manner. The cell adhesion was partially blocked by anti-Mac-1 mAb and inhibition was enhanced when anti-Mac-1 antibodies were combined with glycosaminoglycans, suggesting that cell-surface proteoglycans act cooperatively with Mac-1. PF4 also induced Mac-1-dependent migration of human neutrophils and murine WT, but not Mac-1-deficient macrophages. Coating of Escherichia coli bacteria or latex beads with PF4 enhanced their phagocytosis by macrophages by 4-fold, and this process was blocked by different Mac-1 antagonists. Furthermore, PF4 potentiated phagocytosis by WT, but not Mac-1-deficient macrophages. As determined by biolayer interferometry, PF4 directly bound the MI-domain, the major ligand-binding region of Mac-1, and this interaction was governed by a Kd of 1.3 0.2 M. Using the PF4-derived peptide library, synthetic peptides duplicating the MI-domain recognition sequences and recombinant mutant PF4 fragments, the binding sites for MI-domain were identified in the PF4 segments Cys12–Ser26 and Ala57–Ser70. These results identify PF4 as a ligand for the integrin Mac-1 and suggest that many immune-modulating effects previously ascribed to PF4 are mediated through its interaction with Mac-1.",
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AB - Platelet factor 4 (PF4) is one of the most abundant cationic proteins secreted from -granules of activated platelets. Based on its structure, PF4 was assigned to the CXC family of chemokines and has been shown to have numerous effects on myeloid leukocytes. However, the receptor for PF4 remains unknown. Here, we demonstrate that PF4 induces leukocyte responses through the integrin Mac-1 (M2, CD11b/CD18). Human neutrophils, monocytes, U937 monocytic and HEK293 cells expressing Mac-1 strongly adhered to immobilized PF4 in a concentration-dependent manner. The cell adhesion was partially blocked by anti-Mac-1 mAb and inhibition was enhanced when anti-Mac-1 antibodies were combined with glycosaminoglycans, suggesting that cell-surface proteoglycans act cooperatively with Mac-1. PF4 also induced Mac-1-dependent migration of human neutrophils and murine WT, but not Mac-1-deficient macrophages. Coating of Escherichia coli bacteria or latex beads with PF4 enhanced their phagocytosis by macrophages by 4-fold, and this process was blocked by different Mac-1 antagonists. Furthermore, PF4 potentiated phagocytosis by WT, but not Mac-1-deficient macrophages. As determined by biolayer interferometry, PF4 directly bound the MI-domain, the major ligand-binding region of Mac-1, and this interaction was governed by a Kd of 1.3 0.2 M. Using the PF4-derived peptide library, synthetic peptides duplicating the MI-domain recognition sequences and recombinant mutant PF4 fragments, the binding sites for MI-domain were identified in the PF4 segments Cys12–Ser26 and Ala57–Ser70. These results identify PF4 as a ligand for the integrin Mac-1 and suggest that many immune-modulating effects previously ascribed to PF4 are mediated through its interaction with Mac-1.

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