Effect of mycophenolic acid on TNF alpha-induced expression of cell adhesion molecules in human venous endothelial cells in vitro.
1. Mycophenolic acid (MPA) is an inhibitor of inosine-5'-monophosphate dehydrogenase and therefore interferes with cellular GTP biosynthesis. Recently, MPA has been used as an antiproliferative and immunosuppressive agent. In the present study, the effect of MPA on the expression of the endothelial cell adhesion molecules (CAMs), intercellular (I) CAM-1, vascular (V) CAM-1 and endothelial (E)-selectin, was investigated in tumour necrosis factor-alpha (TNF alpha)-activated cultured human venous endothelial cells (EC). 2. Surface expression of CAMs was measured by flow cytometry and mRNA expression by Northern blot analysis. Transcriptional activation of CAMs by the nuclear factor NF-kappaB was determined by an electromobility shift assay. The function of CAMs was studied by a static adhesion assay with human monocyte-like undifferentiated U937 cells. 3. Pretreatment of TNF alpha- (5 ng ml(-1), 12 h) activated EC with MPA (10 microM, 24 h) increased the binding of U937 cells, which had not been treated with MPA, by approximately 2 fold. MPA-pretreatment of EC did not affect TNF alpha-induced surface expression of ICAM-1. However, VCAM-1 and E-selectin were increased 2-3 fold and remained elevated up to 24 h, by which time TNF alpha-activated control EC had returned to baseline levels of expression. The effect of MPA on the surface expression of CAMs was half-maximal at approximately 1 microM and required > or = 12 h of pretreatment. Guanosine (0.3 mM), a precursor of GTP, did not prevent the effect of MPA on the expression of CAMs in TNF alpha-activated EC. 4. Kinetics of mRNA expression of CAMs mirrored protein expression: mRNA for ICAM-1 was unaffected, whereas TNF alpha-induced mRNA expression for E-selectin and VCAM-1 was prolonged and increased by MPA. This effect was not due to increased transcription mediated by the nuclear transcription factor NF-kappaB. However, half-life for E-selectin mRNA was increased 10 fold by MPA, whereas ICAM-1 mRNA half-life was unchanged. 5. The data demonstrate that apart from its antiproliferative effects on lymphocytes, MPA enhances TNF alpha-induced VCAM-1 and E-selectin surface expression on EC by selectively increasing the mRNA-stability of these cell adhesion molecules. This effect of MPA on EC appears to be independent from inhibition of inosine-5'-monophosphate dehydrogenase.
Hauser
I A
IA
Department of Medicine IV, University of Erlangen/Nürnberg, Nürnberg, F.R.G.
Johnson
D R
DR
Thévenod
F
F
Goppelt-Strübe
M
M
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D006151:Guanosine / Q000494:pharmacology; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D066298:In Vitro Techniques; D009173:Mycophenolic Acid / Q000031:analogs & derivatives / Q000494:pharmacology*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D015854:Up-Regulation; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism
D000903:Antibiotics, Antineoplastic; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007166:Immunosuppressive Agents; D016328:NF-kappa B; D012333:RNA, Messenger; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D006151:Guanosine; D009173:Mycophenolic Acid