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. Author manuscript; available in PMC: 2023 May 19.
Published in final edited form as: Cell Chem Biol. 2022 Feb 28;29(5):811–823.e7. doi: 10.1016/j.chembiol.2022.02.004

Figure 4. Generation and immunogenicity of Aquifex aeolicus Lumazine synthase (AaLS) nanoparticles displaying rsZDIIIs.

Figure 4.

(A) Modelled structural cross-section of AaLS-rsZDIII nanoparticle (AaLS protomers shown in light/dark gray; SpyT/SpyC complex shown in orange; rsZDIII-2.39 shown in blue). (B) Generation of AaLS-rsZDIII nanoparticles by bio-coupling of AaLS-SpC with rsZDIIIs-SpT. SpyC and SpyT moieties spontaneously react to covalently associate. (C) Negative stain electron microscopy of AaLS-rsDIII-2.39 nanoparticle preparations, 50 nm scale bar shown for reference. (D) Mouse immunization and blood collection schedule. (E) D35 and D56 serum reactivity (n=5) for each immunogen group determined by ELISA. Data from a single experiment, performed in triplicate, shown as EC50 values for each animal, including −Log10 (EC50 mean) ± SD for each group. (F) D56 serum FRNT neutralization titers for each immunization group. Data graphed as mean FRNT50 values for each animal (n=5–10) from two independent experiments in triplicate, including Log10 (FRNT50 mean) ± SD for the group. (G) Cross-reactivity of D56 sera elicited by AaLS-ZDIII-WT and AaLS-rsZDIII-2.39 against DENV1–4 EDIIIs. Data from two independent experiments in triplicate, shown as mean EC50 values for each animal (n=3), including −Log10 (EC50 mean) ± SD. (FRNT neutralization and cross-reactivity analysis: 2-way ANOVA with Tukey’s multiple comparisons test: *p< 0.05; **p<0.01; ***p<0.001; ****p<0.0001).