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Figure 1

From: Use of Hemagglutinin Stem Probes Demonstrate Prevalence of Broadly Reactive Group 1 Influenza Antibodies in Human Sera

Figure 1

Design and characterization of HA SS protein. (A) Schematic of the HA SS (bottom) in comparison to HA (top). HA SS was constructed by inserting a GWG linker between residues 42 and 314 of HA1 (red) RBD head, a gp41 post-fusion trimerization motif (green) inserted in place of residues 59 through 93 of HA2 (blue), a GG linker between HA2 and the gp41 HR2 helix and an NGTGGGSG linker between the two gp41 helices. (B) Trimeric and monomeric representation of HA (PDB entry 1RU7) in comparison to the HA SS model. Coloring is respective to panel (A), with the monomeric representation also illustrating the CR6261 epitope as yellow and HA residues which are omitted in the stabilized HA stem as grey. (C) Binding of two bnAb, CR6261 and FI6v3, to WT and SS HA by ELISA. (D) Competition of CR6261 ELISA binding, and (E) pseudotyped neutralization with WT and Δstem (Ile54Arg/Thr49Arg HA2 mutations) HA and HA SS as competitors. HIV gp120 serves as a control. (F) CR6261 (10 µg/mL) ELISA reactivity to WT and ∆stem HA SS probes of different subtypes/strains. H1 HA NC99 and HIV gp120 serve as controls. Error bars represent standard deviation of averaged points in each panel.

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