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

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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

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Keywords

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

ASJC Scopus subject areas

  • Hematology

Cite this

Kameda, H., Morita, I., Handa, M., Kaburaki, J., Yoshida, T., Mimori, T., Murota, S. I., & Ikeda, Y. (1997). Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin. British Journal of Haematology, 97(2), 348-355.