Soluble CD154 does not induce an inflammatory response of endothelial cells.
HUVECs were cultured for 4 hours in medium alone, in a supernatant from thrombin-activated platelets (corresponding to 2 × 108platelets/mL, containing 40 U/mL sCD154), or in a supernatant containing 1000 U/mL sCD154. In parallel, HUVECs were cocultured with the CD154 transfectant P3xTB.A7 alone, or cocultured with the CD154 transfectant P3xTB.A7 in the presence of 1000 U/mL sCD154. Thereafter, the HUVECs were dislodged from the culture dishes and analyzed for expression of the adhesion molecules E-selectin, VCAM-1, and ICAM-1 by flow cytometry. The effects observed with the CD154 transfectant could be fully blocked with CD154-specific mAb TRAP1 (not shown and Henn et al12).
