Effect of mycophenolic acid on TNFα‐induced expression of cell adhesion molecules in human venous endothelial cells in vitro

Abstract

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‐α (TNFα)‐activated cultured human venous endothelial cells (EC). 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‐κB 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. Pretreatment of TNFα‐ (5 ng ml−1, 12 h) activated EC with MPA (10 μM, 24 h) increased the binding of U937 cells, which had not been treated with MPA, by ∼amp;2 fold. MPA‐pretreatment of EC did not affect TNFα‐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 TNFα‐activated control EC had returned to baseline levels of expression. The effect of MPA on the surface expression of CAMs was half‐maximal at ∼amp;1 μM and required 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 TNFα‐activated EC. Kinetics of mRNA expression of CAMs mirrored protein expression: mRNA for ICAM‐1 was unaffected, whereas TNFα‐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‐κB. However, half‐life for E‐selectin mRNA was increased 10 fold by MPA, whereas ICAM‐1 mRNA half‐life was unchanged. The data demonstrate that apart from its antiproliferative effects on lymphocytes, MPA enhances TNFα‐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.

Impact of Different Immunosuppressive Regimens on Antigen-Presenting Blood Cells in Kidney Transplant Patients

“…RANTES, was synergistically increased, indicating a possible proinflammatory effect of MMF on RANTES release [9]. In similar manner, in vitro data showed that apart from antiproliferative effects, MMF enhances TNF-induced expression of VCAM-1 and E-selectin on human endothelial cells [10]. The clinical significance of a these findings remains unclear, however, and should be mentioned in association with the data presented here.…”

Effects of mycophenolic acid on endothelial cells

“…These results are partly in agreement with the finding of Raab M et al [10] who demonstrated an inhibitory effect of MPA on TNFa-stimulated expression of ICAM-1, VCAM-1 and E-selectin in HUVECs. In contrast, Hauser IA et al [22] showed that MPA increased TNFastimulated expression of VCAM-1 and E-selectin, but not of ICAM-1 in HUVECs.…”

NF-κB balances vascular regression and angiogenesis via chromatin remodeling and NFAT displacement

“…9 We used promoter arrays to measure the activity of multiple TFs in remodeling ECs after exposure to the angioinhibitory protein pigment epithelial-derived factor (PEDF). We chose nuclear factor-B (NF-B) because it is implicated in angiogenesis, 10 drives Fas ligand (FasL) expression, 11 and is activated by PEDF in non-ECs. 12 NF-B can play 2 opposing roles in tumorigenesis.…”