Antagonistic antibody prevents toll-like receptor 2–driven lethal shock-like syndromes
J. Clin. Invest. Guangxun Meng, et al. 113:1473 doi:10.1172/JCI20762 [Go to this article.]

Figure 4
Molecular analysis of the effects of mAb T2.5 on TLR2ECD-P₃CSK₄ interaction. (A and B) Binding of recombinant TLR2ECD-Fc fusion protein (T2EC, positive controls) to immobilized P₃CSK₄ upon preincubation with T2.5 (T2EC + T2.5) at different molar excesses (A, ∞1, ∞3.3, ∞10) or with an isotype-matched control mAb (T2EC + con) at tenfold molar excess only (B, ∞10). Binding was continuously monitored in an SPR biosensor device, and amounts of antibodies used to gain high molecular excess over T2EC (coincubation) were applied alone as negative controls (A, T2.5; B, Con). Response units at 300 seconds are a measure for P₃CSK₄-binding capacities of T2EC and T2EC plus mAb. (C) For analysis of approximate localization of T2.5 epitope within the TLR2ECD, a mutant human TLR2 construct lacking the N-terminal third of the LRR-rich ECD (hTLR2-mutH) was used for NF-κB–dependent luciferase assay upon transient transfection, preincubation with mAb (T2.5, conT2), and P₃CSK₄ challenge (black bars). Absence of mAb treatment (No mAb) and/or of P₃CSK₄ challenge (white bars), and empty vector (Vector), represent respective controls.