Introduction Dysregulation of Toll-like receptor 4 (TLR4) signaling via numerous exogenous and endogenous ligands appears to play an underlying role in the pathogenesis of multiple inflammatory diseases. Since multiple TLR4 ligands may be upregulated in a single disease condition, blockade of TLR4 activation via a ligand-independent mechanism is an attractive strategy for disease intervention. A ligand-independent anti-TLR4 neutralizing antibody evaluated as a novel strategy for TLR4 neutralization. Methods An anti-human TLR4 mAb, NI-0101, was generated and shown to bind to human TLR4 in a region involved in receptor dimerization. Using exogenous, endogenous or chemical ligands, the capacity to block TLR4 activation was evaluated. Subsequently, NI-0101 was tested in islet-induced immune cell activation. Furthermore, the corresponding mouse surrogate antibody was tested in a laboratory model of islet transplantation. Results Due to inhibition of TLR4 dimerization, NI-0101 efficiently blocked the activation of TLR4 by different ligands. Consistent with its ligand-independent mechanism of action, a similar inhibitory potency of NI-0101 was observed at different concentrations of the activating ligand. In addition, NI-0101 could block human islet-induced human immune cell activation, further confirming the ability of NI-0101 to block TLR4 activation independently of the ligand. To determine the effects of NI-0101 in vivo , we used the mouse surrogate antibody of NI-0101, 5E3. 5E3 effectively protected grafted islets in a mouse model of islet transplantation. Conclusion The therapeutic anti-human TLR4 mAb, NI-0101, has the capacity to interfere with not only LPS but also signaling of TLR4 through endogenous and chemical ligands. When used in an in vitro model of islet-induced immune cell activation, NI-0101 blocked immune cell activation. Furthermore, we demonstrated that anti-TLR4 mAb is efficient in protecting grafted islets in a mouse islet transplantation model. Taken together, these data promote ligand-independent blockade of TLR4 activation as a promising strategy in blocking disease pathogenesis.
Read full abstract