VARIABILITY IN E-SELECTIN EXPRESSION, mRNA LEVELS AND sE-SELECTIN RELEASE BETWEEN ENDOTHELIAL CELL LINES AND PRIMARY ENDOTHELIAL CELLS

Authors

  • H.F Galley,

    Corresponding author
    1. Academic Unit of Anaesthesia and Intensive Care, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen, AB25 2ZD, U.K.
      To whom correspondence should be addressed. E-mail:h.f.galley@abdn.ac.uk
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  • M.G Blaylock,

    1. Academic Unit of Anaesthesia and Intensive Care, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen, AB25 2ZD, U.K.
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  • A.M Dubbels,

    1. Academic Unit of Anaesthesia and Intensive Care, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen, AB25 2ZD, U.K.
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  • N.R Webster

    1. Academic Unit of Anaesthesia and Intensive Care, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen, AB25 2ZD, U.K.
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To whom correspondence should be addressed. E-mail:h.f.galley@abdn.ac.uk

Abstract

Endothelial cell lines express markers and are assumed to exhibit other endothelial cell responses. We investigated E-selectin expression from human umbilical vein endothelial cells, the spontaneously transformed ECV304 line and the hybrid line EA.hy926 by flow cytometry and immunofluorescence, mRNA and soluble E-selectin release. In cells exposed to tumour necrosis factor α (TNF-α) and interleukin-1β (IL-1β), median (range) percentage of E-selectin-positive HUVECs increased from 1.6(0.9–6.2)% to 91.4(83.0–96.1)%, (P=0.001) using flow cytometry. In contrast, E-selectin expression by ECV304 and EA.hy926 cell lines was 100-fold lower. E-selectin mRNA was detectable after 2h, maximal at 6h in HUVECs and undetectable in EA.hy926 and ECV304 cell lines after exposure to TNF-α/IL-1β. sE-selectin accumulation increased (P=0.004) in HUVECs only. Neutrophil adherence to ECV304 and EA.hy926 cells was poor compared to HUVECs (P=0.004). The cell lines ECV304 and EA.hy926 do not exhibit normal endothelium expression of E-selectin, and may not be appropriate for studies of adhesion.

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