RESUMO
AIM: To identify the inhibition of TNF-induced NF-kappaB nuclear translocation by three anti-human TNF mAbs, D2, E6 and F6. METHODS: TNF solutions were pretreated with mAbs D2, E6 and F6 as well as control mAb at 37 degrees Celsius for 1 h, respectively, and then they were added to ECV304 cell cultures. After 1 hour, the cells were harvested and nuclear proteins were extracted. The NF-kappaB activity in nuclear extract was detected by electrophoretic mobility shift assay (EMSA). RESULTS: All of the three anti-TNF mAbs could inhibit TNF-induced NF-kappaB nuclear translocation in a dose-dependent manner. At the concentrations of 10 mg/L and 0.1 mg/L, the inhibition rates of mAb D2, E6 and F6 were 94.2% and 75.1%, 64.9% and 28.6%, 70.3% and 49.5% respectively, while the inhibition rate of control mAb was only 20.0% and 11.1%. CONCLUSION: mAbs D2, E6 and F6 can specifically inhibit TNF-induced NF-kappaB nuclear translocation, which lays the foundation for preparation of therapeutic chimeric anti-human TNF antibody for treatment of infectious and autoimmune diseases.