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

AIM: NE-κB, 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-IκBαM vector to inhibit NE-κB activation and investigate the effect of pCMV-IκBαM in inhibition of T cells adhesion to endothelial cells.METHODS: The NF-κB activity was detected with pNF-κB 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-I~BczMin in hibition of T cells adhesion to endothelial cells.RESULTS: We could find that NF-~B activity is inhibited by over-expression of non-degraded IκBα 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-IκBαM vector.CONCLUSION: Our results indicate that the pCMV-IκBαM, which inhibit the activity of NF-κB through over-expression of non-degraded IκBα protein, can be used for gene therapy in diseases involving NF-κB activation abnormally like organ transplantation via inhibiting cell adhesion.

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

AIM: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.