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. Author manuscript; available in PMC: 2018 Mar 8.
Published in final edited form as: Sci Transl Med. 2017 Mar 8;9(380):eaaf5347. doi: 10.1126/scitranslmed.aaf5347

Fig. 8. Co-formulation of semisynthetic ST8 glycoconjugates and Prevnar 13.

Fig. 8

(A) Adsorption of ST8 glycoconjugates to Prevnar 13, as assessed by flow cytometry. ST8 glycoconjugates were adsorbed to Prevnar 13 particles using 25%, 100% or 400% of a full dose equivalent, and detected by flow cytometry using mAb 1H8 and an Alexa Fluor 635-labeled detection antibody. Controls are non-treated and CRM197-treated Prevnar 13 particles. Data are means + SD of the median fluorescence intensities of n = 3 independent experiments. (B) Immunization strategy. Rabbits (n = 3 per group) were s.c. immunized 3 times with Prevnar 13 with or without co-adsorbed CRM197 glycoconjugates. Serum was collected at day 35. (C–D) Immune response to ST8 CPS as assessed by polysaccharide ELISA. (C) Evaluation of ST8 CPS binding by rabbit sera at day 35 (1:200 dilution). Data are medians ± range from n = 3 individual rabbits per group. P values are determined by one-tailed Mann-Whitney U test. (D) Comparison of ST8 CPS binding of rabbit sera from different groups. Data are means ± SD of n = 3 rabbits per group. (E) Comparison of opsonophagocytic killing of ST8 pneumococci by pooled rabbit sera. Data are means + SD of cfu reduction relative to control wells of n = 3 incubation wells as biological replicates at a 1:32 serum dilution. P values are determined by one-tailed, unpaired t test with Welch’s correction. (F) Comparison of opsonophagocytic killing by pooled rabbit sera and human reference serum 007sp at different serum dilutions. Data are means ± SD of cfu reduction relative to control wells of n = 3 incubation wells. Data in (E) and (F) are from one representative out of 3 independent experiments.