Inhibition of PMA-induced endothelial cell activation and adhesion by over-expression of domain negative IkappaBalpha protein

• Published in: World journal of gastroenterology : WJG 2005-05, Vol.11 (20), p.3080
• Main Authors: Wei, Jian-Feng, Sun, Ke, Xu, Shi-Guo, Xie, Hai-Yang, Zheng, Shu-Sen
• Format: Article
• Language: English
• Subjects: Cell Adhesion; Cell Line; Endothelial Cells - drug effects; Endothelial Cells - immunology; Endothelium, Vascular - cytology; Graft Rejection - immunology; Humans; I-kappa B Proteins - biosynthesis; NF-KappaB Inhibitor alpha; T-Lymphocytes - cytology; T-Lymphocytes - immunology; Tetradecanoylphorbol Acetate - pharmacology
• ISSN: 1007-9327

NF-kappaB, regulate the expression of cytokine-inducible genes involving immune and inflammatory responses, will be potential therapy approach for allograft from rejection. In this study, we use pCMV- IkappaBalphaM vector to inhibit NF-kappaB activation and investigate the effect of pCMV- IkappaBalphaM in inhibition of T cells adhesion to endothelial cells. The NF-kappaB activity was detected with pNF-kappaB reporter gene and electrophoretic mobility shift assay. Expression of cell surface molecules was detected by RT-PCR and flow cytometer. The cell-cell adhesion assay was performed to determine the effect of pCMV-IkappaBalphaM in inhibition of T cells adhesion to endothelial cells. We could find that NF-kappaB activity is inhibited by over-expression of non-degraded IkappaBalpha protein. Expression of adhesion molecules like ICAM-1, VCAM-1, and P-selectin as well as cell-cell adhesion were inhibited significantly by transfection of the pCMV- IkappaBalphaM vector. Our results indicate that the pCMV- IkappaBalphaM, which inhibit the activity of NF-kappaB through over-expression of non-degraded IkappaBalpha protein, can be used for gene therapy in diseases involving NF-kappaB activation abnormally like organ transplantation via inhibiting cell adhesion.