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. 2020 Oct 16;11:577823. doi: 10.3389/fimmu.2020.577823

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Copyright © 2020 Wang, Bazin-Lee, Evans, Casella and Mitchell

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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ML3 and ML4A function as competitive antagonists of human TLR4. MPL adjuvant^® components ML3 and ML4A were tested for competitive antagonism of human TLR4 using reporter cells. The agonists (A) PHAD and (B) 3D-6A-PHAD were diluted in half-log steps from a peak concentration of 10 μM in a mixture containing a fixed concentration of ML3 (3 μM), ML4A (3 μM) or Lipid IVa (0.1 μM). Values shown are averages ± SD of secreted alkaline phosphatase from two independent experiments, each performed in duplicate or triplicate and normalized by setting 100% = PHAD or 3D-6A-PHAD alone (3 μM). Shaded regions indicate 99% confidence intervals within which the true population means should occur 99% of the time. Agonist curves in the presence of a fixed amount of ML3 or ML4A reached the same top dose plateaus as for agonist alone indicating both are competitive antagonists of human TLR4.