Expression of Neutral Glycosphingolipids in Cytokine-Stimulated Human Endothelial Cells

T. Zemunik^1*, A. Markotić², and A. Marusić³

¹Department of Biology, School of Medicine, Split University, Soltanska 2, 21000 Split, Croatia; fax: (++385-21) 465-212; E-mail: tzemunik@bsb.mefst.hr

²Department of Biochemistry, School of Medicine, Split University, Soltanska 2, 21000 Split, Croatia; fax: (++385-21) 557-625; E-mail: markotic@bsb.mefst.hr

³Department of Anatomy and Croatian Institute for Brain Research, University School of Medicine, Salata 3, 10000 Zagreb, Croatia; fax: (++385-01) 01459-0222; E-mail: marusica@mef.hr

^* To whom correspondence should be addressed.

Received July 11, 2003; Revision received November 25, 2003

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We compared immunohistochemical distribution of glycosphingolipids globotriosylceramide (GbOse₃Cer) and globotetraosylceramide (GbOse₄Cer) with that of E-selectin on human umbilical vein endothelial cells (HUVEC) stimulated with tumor necrosis factor (TNF)-alpha. HUVECs activated by TNF-alpha were characterized by the highest expression of E-selectin and greatest adhesion of HL-60 cells as well compared to stimulation with interleukin-1beta or lipopolysaccharide. HUVECs activated by TNF-alpha also stained intensely with globoside antibodies, especially with the GbOse₃Cer-directed one, staining being redistributed in a concentration-dependent manner. These results indicate the possible role of GbOse₃Cer and GbOse₄Cer in immune effector mechanisms of endothelium such as adhesion.

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KEY WORDS: endothelium, glycosphingolipids, E-selectin, TNF-alpha, immunohistochemistry

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