Impaired induction of adhesion molecule expression in immortalized endothelial cells leads to functional defects in...

Immortalization should overcome the problem of short lifespan and difficult culture of endothelial cells that limited their use in functional studies. We used four different immortalized endothelial cell lines to study dynamic interactions with lymphocytes. Surprisingly, tumor necrosis factor (TNF)alpha-stimulated human umbilical vein endothelial cells (HUVECs) or human dermal microvascular endothelial cells (HDMECs) readily supported rolling and binding of lymphocytes, whereas none of the immortalized cell lines did. As rolling interactions are primarily mediated by selectins and vascular cell adhesion molecule (VCAM)-1, the endothelial cells were analyzed regarding expression of selectins and other adhesion molecules. Interestingly, cell surface expression of E-selectin could only be detected on HUVEC and HDMEC. Immunocytochemistry showed that some immortalized endothelial cells expressed E-selectin intracellularly following TNFalpha stimulation, suggesting translation but defective post-translational processing or transport of the molecule. In contrast, other immortalized cell lines did not have detectable levels of E-selectin mRNA, suggesting impaired transcription. VCAM-1 could only be induced on normal and human placental microvascular endothelial cell-A2 endothelial cells, whereas all cell lines expressed intercellular adhesion molecule-1 following TNF stimulation. The immortalized endothelial cells tested here have lost functions that are required for dynamic interactions with immune cells and that are common to primary endothelial cells.