[The inhibition of TNF-induced NF-kappaB nuclear translocation in ECV304 cells by mAbs against human TNF].
OBJECTIVE
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%.


CONCLUSIONS
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.
Zhu
Can-sheng
CS
Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Ouyang
Wei-ming
WM
Liu
Xue-song
XS
Liang
Xiao-cong
XC
Song
Chao-jun
CJ
Jin
Bo-quan
BQ
D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects*; D048069:Tumor Necrosis Factors / Q000276:immunology*; D014471:Umbilical Veins / Q000166:cytology
D000911:Antibodies, Monoclonal; D016328:NF-kappa B; D048069:Tumor Necrosis Factors