Abstract
Tumor necrosis factor (TNF)-α genetically fused to the carboxyl terminus of a single-chain Fv (ScFv) antibody specific for the human HER2/neu (anti-HER2/neu ScFv-TNF-α) forms a homotrimeric structure that retains both TNF-α activity and the ability to bind HER2/neu. In contrast to anti-HER2/neu IgG3, anti-HER2/neu ScFv-TNF-α induces potent HER2/neu signaling, activating the downstream mitogen-activated protein kinase (MAPK) and Akt pathways in SKBR3 cells. Activation of MAPK and Akt by anti-HER2/neu ScFv-TNF-α inhibited the apoptosis of SKBR3 cells induced by actinomycin D. Remarkably, anti-HER2/neu ScFv-TNF-α facilitated the repair of injured epithelia. Accelerated wound healing required binding to HER2/neu but not TNF-α activity since anti-HER2/neu ScFv-TNF-α (S147Y), containing a mutant TNF-α with significantly decreased biological activity, demonstrated equivalent ability to facilitate wound healing and soluble HER2/neu inhibited the effect. These results suggest that trimeric anti-HER2/neu ScFv has the potential to facilitate wound healing. In addition, fusion with TNF-α provides a novel approach to producing polymeric antibodies.
- The American Society for Pharmacology and Experimental Therapeutics
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