Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin

Hideto Kameda, Ikuo Morita, Makoto Handa, Junichi Kaburaki, Tadashi Yoshida, Tsuneyo Mimori, Sei Itsu Murota, Yasuo Ikeda

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

P-selectin (GMP-140, PADGEM, CD62P) is a cell adhesion receptor which is believed to play an important role in inflammatory diseases by supporting leucocyte rolling. P-selectin is located on the granule membrane of Weibel- Palade bodies in resting endothelial cells and is expressed on the cell surface during cellular activation with various stimulators such as thrombin. Thereafter, P-selectin is internalized and sorted to the Golgi region and Weibel-Palade bodies again. However, whether P-selectin is re-expressed upon subsequent cellular stimulation has, to date, been unclear. To address this question, we measured the cellular content and surface expression of P- selectin, using indirect immunofluorescence and confocal laser cytometry. Surface expression of P-selectin reached a maximum < 2 min after thrombin stimulation and declined to basal levels after 18O min. Rechallenge with thrombin induced rapid surface re-expression of P-selectin, which was independent of de novo protein synthesis, since cycloheximide did not inhibit re-expression. Moreover, re-expressed P-selectin supported the adherence of HL60 promyelocytic cells. These results clearly demonstrated that functional P-selectin molecule was recycled after repeated stimulation with thrombin, raising the possibility that P-selectin is involved in chronic inflammation.

Original languageEnglish
Pages (from-to)348-355
Number of pages8
JournalBritish Journal of Haematology
Volume97
Issue number2
Publication statusPublished - 1997

Fingerprint

P-Selectin
Thrombin
Endothelial Cells
Weibel-Palade Bodies
Leukocyte Rolling
HL-60 Cells
Cycloheximide
Indirect Fluorescent Antibody Technique
Cell Adhesion
Lasers

Keywords

  • adhesion
  • chronic inflammation
  • degranulation
  • protein synthesis
  • recycling

ASJC Scopus subject areas

  • Hematology

Cite this

Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin. / Kameda, Hideto; Morita, Ikuo; Handa, Makoto; Kaburaki, Junichi; Yoshida, Tadashi; Mimori, Tsuneyo; Murota, Sei Itsu; Ikeda, Yasuo.

In: British Journal of Haematology, Vol. 97, No. 2, 1997, p. 348-355.

Research output: Contribution to journalArticle

Kameda, H, Morita, I, Handa, M, Kaburaki, J, Yoshida, T, Mimori, T, Murota, SI & Ikeda, Y 1997, 'Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin', British Journal of Haematology, vol. 97, no. 2, pp. 348-355.
Kameda, Hideto ; Morita, Ikuo ; Handa, Makoto ; Kaburaki, Junichi ; Yoshida, Tadashi ; Mimori, Tsuneyo ; Murota, Sei Itsu ; Ikeda, Yasuo. / Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin. In: British Journal of Haematology. 1997 ; Vol. 97, No. 2. pp. 348-355.
@article{974872f13a0a494fa6d04d5439ba6124,
title = "Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin",
abstract = "P-selectin (GMP-140, PADGEM, CD62P) is a cell adhesion receptor which is believed to play an important role in inflammatory diseases by supporting leucocyte rolling. P-selectin is located on the granule membrane of Weibel- Palade bodies in resting endothelial cells and is expressed on the cell surface during cellular activation with various stimulators such as thrombin. Thereafter, P-selectin is internalized and sorted to the Golgi region and Weibel-Palade bodies again. However, whether P-selectin is re-expressed upon subsequent cellular stimulation has, to date, been unclear. To address this question, we measured the cellular content and surface expression of P- selectin, using indirect immunofluorescence and confocal laser cytometry. Surface expression of P-selectin reached a maximum < 2 min after thrombin stimulation and declined to basal levels after 18O min. Rechallenge with thrombin induced rapid surface re-expression of P-selectin, which was independent of de novo protein synthesis, since cycloheximide did not inhibit re-expression. Moreover, re-expressed P-selectin supported the adherence of HL60 promyelocytic cells. These results clearly demonstrated that functional P-selectin molecule was recycled after repeated stimulation with thrombin, raising the possibility that P-selectin is involved in chronic inflammation.",
keywords = "adhesion, chronic inflammation, degranulation, protein synthesis, recycling",
author = "Hideto Kameda and Ikuo Morita and Makoto Handa and Junichi Kaburaki and Tadashi Yoshida and Tsuneyo Mimori and Murota, {Sei Itsu} and Yasuo Ikeda",
year = "1997",
language = "English",
volume = "97",
pages = "348--355",
journal = "British Journal of Haematology",
issn = "0007-1048",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin

AU - Kameda, Hideto

AU - Morita, Ikuo

AU - Handa, Makoto

AU - Kaburaki, Junichi

AU - Yoshida, Tadashi

AU - Mimori, Tsuneyo

AU - Murota, Sei Itsu

AU - Ikeda, Yasuo

PY - 1997

Y1 - 1997

N2 - P-selectin (GMP-140, PADGEM, CD62P) is a cell adhesion receptor which is believed to play an important role in inflammatory diseases by supporting leucocyte rolling. P-selectin is located on the granule membrane of Weibel- Palade bodies in resting endothelial cells and is expressed on the cell surface during cellular activation with various stimulators such as thrombin. Thereafter, P-selectin is internalized and sorted to the Golgi region and Weibel-Palade bodies again. However, whether P-selectin is re-expressed upon subsequent cellular stimulation has, to date, been unclear. To address this question, we measured the cellular content and surface expression of P- selectin, using indirect immunofluorescence and confocal laser cytometry. Surface expression of P-selectin reached a maximum < 2 min after thrombin stimulation and declined to basal levels after 18O min. Rechallenge with thrombin induced rapid surface re-expression of P-selectin, which was independent of de novo protein synthesis, since cycloheximide did not inhibit re-expression. Moreover, re-expressed P-selectin supported the adherence of HL60 promyelocytic cells. These results clearly demonstrated that functional P-selectin molecule was recycled after repeated stimulation with thrombin, raising the possibility that P-selectin is involved in chronic inflammation.

AB - P-selectin (GMP-140, PADGEM, CD62P) is a cell adhesion receptor which is believed to play an important role in inflammatory diseases by supporting leucocyte rolling. P-selectin is located on the granule membrane of Weibel- Palade bodies in resting endothelial cells and is expressed on the cell surface during cellular activation with various stimulators such as thrombin. Thereafter, P-selectin is internalized and sorted to the Golgi region and Weibel-Palade bodies again. However, whether P-selectin is re-expressed upon subsequent cellular stimulation has, to date, been unclear. To address this question, we measured the cellular content and surface expression of P- selectin, using indirect immunofluorescence and confocal laser cytometry. Surface expression of P-selectin reached a maximum < 2 min after thrombin stimulation and declined to basal levels after 18O min. Rechallenge with thrombin induced rapid surface re-expression of P-selectin, which was independent of de novo protein synthesis, since cycloheximide did not inhibit re-expression. Moreover, re-expressed P-selectin supported the adherence of HL60 promyelocytic cells. These results clearly demonstrated that functional P-selectin molecule was recycled after repeated stimulation with thrombin, raising the possibility that P-selectin is involved in chronic inflammation.

KW - adhesion

KW - chronic inflammation

KW - degranulation

KW - protein synthesis

KW - recycling

UR - http://www.scopus.com/inward/record.url?scp=0030922560&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030922560&partnerID=8YFLogxK

M3 - Article

VL - 97

SP - 348

EP - 355

JO - British Journal of Haematology

JF - British Journal of Haematology

SN - 0007-1048

IS - 2

ER -