Functional and structural characterization of the eosinophil P-selectin ligand

F A Symon, M B Lawrence, M L Williamson, Garry Michael Walsh, A J Wardlaw

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

Our recent studies have indicated an important role for P-selectin in eosinophil adhesion, We have therefore compared eosinophil and neutrophil binding with nasal polyp endothelium as well as purified P-selectin. We have also compared the structure and expression of the eosinophil and neutrophil P-selectin ligands. Using the frozen section assay, eosinophils bound to 2-fold more blood vessels within the nasal polyp tissue than neutrophils. Up to 10-fold more eosinophils than neutrophils bound per unit length of endothelium, Neutrophil and eosinophil binding was inhibited by a mAb against P-selectin and a P-selectin chimera which binds to the P-selectin ligand. Eosinophils bound with approximately 2-fold greater avidity to purified P-selectin under now conditions, Using SDS-PAGE we characterized the eosinophil P-selectin ligand as a sialylated, homodimeric glycoprotein consistent with the known structure of PSGL-1. However, expression of PSGL-1 by eosinophils was significantly greater than on neutrophils. The eosinophil ligand had a calculated molecular mass by SDS-PACE of approximately 10 kDa greater than the neutrophil ligand, which was not due to differences in N-glycosylation. Eosinophils expressed the 15-decapeptide repeat form of PSGL-1 compared with neutrophils that have the 16-decapeptide repeat form, The increased binding of eosinophils, compared with neutrophils, to P-selectin in both an ex-vivo and in vitro assay suggests that P-selectin may have a role directing the specific migration of eosinophils in diseases such as asthma. The increased avidity may be due to increased expression of PSGL-1 by eosinophils, differences in the peptide backbone, or post-translational modifications.

Original languageEnglish
Pages (from-to)1711-1719
Number of pages9
JournalThe Journal of Immunology
Volume157
Issue number4
Publication statusPublished - 15 Aug 1996

Keywords

  • VASCULAR ENDOTHELIAL-CELLS
  • INTERCELLULAR-ADHESION MOLECULE-1
  • GLYCOPROTEIN LIGAND
  • HUMAN-LEUKOCYTES
  • BRONCHIAL-MUCOSA
  • AMINO-TERMINUS
  • EXPRESSION
  • NEUTROPHILS
  • RECEPTOR
  • MECHANISMS

Cite this

Symon, F. A., Lawrence, M. B., Williamson, M. L., Walsh, G. M., & Wardlaw, A. J. (1996). Functional and structural characterization of the eosinophil P-selectin ligand. The Journal of Immunology, 157(4), 1711-1719.

Functional and structural characterization of the eosinophil P-selectin ligand. / Symon, F A ; Lawrence, M B ; Williamson, M L ; Walsh, Garry Michael; Wardlaw, A J .

In: The Journal of Immunology, Vol. 157, No. 4, 15.08.1996, p. 1711-1719.

Research output: Contribution to journalArticle

Symon, FA, Lawrence, MB, Williamson, ML, Walsh, GM & Wardlaw, AJ 1996, 'Functional and structural characterization of the eosinophil P-selectin ligand', The Journal of Immunology, vol. 157, no. 4, pp. 1711-1719.
Symon FA, Lawrence MB, Williamson ML, Walsh GM, Wardlaw AJ. Functional and structural characterization of the eosinophil P-selectin ligand. The Journal of Immunology. 1996 Aug 15;157(4):1711-1719.
Symon, F A ; Lawrence, M B ; Williamson, M L ; Walsh, Garry Michael ; Wardlaw, A J . / Functional and structural characterization of the eosinophil P-selectin ligand. In: The Journal of Immunology. 1996 ; Vol. 157, No. 4. pp. 1711-1719.
@article{ee909a709ad941298aa3127d28273162,
title = "Functional and structural characterization of the eosinophil P-selectin ligand",
abstract = "Our recent studies have indicated an important role for P-selectin in eosinophil adhesion, We have therefore compared eosinophil and neutrophil binding with nasal polyp endothelium as well as purified P-selectin. We have also compared the structure and expression of the eosinophil and neutrophil P-selectin ligands. Using the frozen section assay, eosinophils bound to 2-fold more blood vessels within the nasal polyp tissue than neutrophils. Up to 10-fold more eosinophils than neutrophils bound per unit length of endothelium, Neutrophil and eosinophil binding was inhibited by a mAb against P-selectin and a P-selectin chimera which binds to the P-selectin ligand. Eosinophils bound with approximately 2-fold greater avidity to purified P-selectin under now conditions, Using SDS-PAGE we characterized the eosinophil P-selectin ligand as a sialylated, homodimeric glycoprotein consistent with the known structure of PSGL-1. However, expression of PSGL-1 by eosinophils was significantly greater than on neutrophils. The eosinophil ligand had a calculated molecular mass by SDS-PACE of approximately 10 kDa greater than the neutrophil ligand, which was not due to differences in N-glycosylation. Eosinophils expressed the 15-decapeptide repeat form of PSGL-1 compared with neutrophils that have the 16-decapeptide repeat form, The increased binding of eosinophils, compared with neutrophils, to P-selectin in both an ex-vivo and in vitro assay suggests that P-selectin may have a role directing the specific migration of eosinophils in diseases such as asthma. The increased avidity may be due to increased expression of PSGL-1 by eosinophils, differences in the peptide backbone, or post-translational modifications.",
keywords = "VASCULAR ENDOTHELIAL-CELLS, INTERCELLULAR-ADHESION MOLECULE-1, GLYCOPROTEIN LIGAND, HUMAN-LEUKOCYTES, BRONCHIAL-MUCOSA, AMINO-TERMINUS, EXPRESSION, NEUTROPHILS, RECEPTOR, MECHANISMS",
author = "Symon, {F A} and Lawrence, {M B} and Williamson, {M L} and Walsh, {Garry Michael} and Wardlaw, {A J}",
year = "1996",
month = "8",
day = "15",
language = "English",
volume = "157",
pages = "1711--1719",
journal = "The Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "4",

}

TY - JOUR

T1 - Functional and structural characterization of the eosinophil P-selectin ligand

AU - Symon, F A

AU - Lawrence, M B

AU - Williamson, M L

AU - Walsh, Garry Michael

AU - Wardlaw, A J

PY - 1996/8/15

Y1 - 1996/8/15

N2 - Our recent studies have indicated an important role for P-selectin in eosinophil adhesion, We have therefore compared eosinophil and neutrophil binding with nasal polyp endothelium as well as purified P-selectin. We have also compared the structure and expression of the eosinophil and neutrophil P-selectin ligands. Using the frozen section assay, eosinophils bound to 2-fold more blood vessels within the nasal polyp tissue than neutrophils. Up to 10-fold more eosinophils than neutrophils bound per unit length of endothelium, Neutrophil and eosinophil binding was inhibited by a mAb against P-selectin and a P-selectin chimera which binds to the P-selectin ligand. Eosinophils bound with approximately 2-fold greater avidity to purified P-selectin under now conditions, Using SDS-PAGE we characterized the eosinophil P-selectin ligand as a sialylated, homodimeric glycoprotein consistent with the known structure of PSGL-1. However, expression of PSGL-1 by eosinophils was significantly greater than on neutrophils. The eosinophil ligand had a calculated molecular mass by SDS-PACE of approximately 10 kDa greater than the neutrophil ligand, which was not due to differences in N-glycosylation. Eosinophils expressed the 15-decapeptide repeat form of PSGL-1 compared with neutrophils that have the 16-decapeptide repeat form, The increased binding of eosinophils, compared with neutrophils, to P-selectin in both an ex-vivo and in vitro assay suggests that P-selectin may have a role directing the specific migration of eosinophils in diseases such as asthma. The increased avidity may be due to increased expression of PSGL-1 by eosinophils, differences in the peptide backbone, or post-translational modifications.

AB - Our recent studies have indicated an important role for P-selectin in eosinophil adhesion, We have therefore compared eosinophil and neutrophil binding with nasal polyp endothelium as well as purified P-selectin. We have also compared the structure and expression of the eosinophil and neutrophil P-selectin ligands. Using the frozen section assay, eosinophils bound to 2-fold more blood vessels within the nasal polyp tissue than neutrophils. Up to 10-fold more eosinophils than neutrophils bound per unit length of endothelium, Neutrophil and eosinophil binding was inhibited by a mAb against P-selectin and a P-selectin chimera which binds to the P-selectin ligand. Eosinophils bound with approximately 2-fold greater avidity to purified P-selectin under now conditions, Using SDS-PAGE we characterized the eosinophil P-selectin ligand as a sialylated, homodimeric glycoprotein consistent with the known structure of PSGL-1. However, expression of PSGL-1 by eosinophils was significantly greater than on neutrophils. The eosinophil ligand had a calculated molecular mass by SDS-PACE of approximately 10 kDa greater than the neutrophil ligand, which was not due to differences in N-glycosylation. Eosinophils expressed the 15-decapeptide repeat form of PSGL-1 compared with neutrophils that have the 16-decapeptide repeat form, The increased binding of eosinophils, compared with neutrophils, to P-selectin in both an ex-vivo and in vitro assay suggests that P-selectin may have a role directing the specific migration of eosinophils in diseases such as asthma. The increased avidity may be due to increased expression of PSGL-1 by eosinophils, differences in the peptide backbone, or post-translational modifications.

KW - VASCULAR ENDOTHELIAL-CELLS

KW - INTERCELLULAR-ADHESION MOLECULE-1

KW - GLYCOPROTEIN LIGAND

KW - HUMAN-LEUKOCYTES

KW - BRONCHIAL-MUCOSA

KW - AMINO-TERMINUS

KW - EXPRESSION

KW - NEUTROPHILS

KW - RECEPTOR

KW - MECHANISMS

M3 - Article

VL - 157

SP - 1711

EP - 1719

JO - The Journal of Immunology

JF - The Journal of Immunology

SN - 0022-1767

IS - 4

ER -