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. 2024 Jun 7;15:4857. doi: 10.1038/s41467-024-48721-3

Fig. 6. Immunisation with combinations of RH5 and R78C improves on immunisation with RH5 alone.

Fig. 6

Wistar rats were immunised on days 0, 28 and 56. A terminal bleed was taken on day 70 (post-third dose). Combination (e.g., “R78C + RH5”) groups were given at equimolar ratios of 9 µg + 10 µg antigen respectively, whereas “mini” groups (e.g., “RCR-78 mini”) were given as 20 µg of the pre-formed complex. Day 70 serum IgG ELISA data (reported in µg/mL) showed against full-length (A, B) RH5 (red), (C, D) CyRPA (blue), and (E, F) RIPR (green). Dotted line: median antigen-specific IgG response from a reference group of single antigen immunised animals. A summary of statistical analysis can be found in Table S3. N = 6 animals per group (G) Single-cycle GIA assays. Total IgG, purified from day 70 serum samples, was titrated in the GIA assay (see Supplementary Fig. 8B). GIA at 1 mg/mL total purified IgG was interpolated. The dotted line indicates the median GIA for the RH5 alone group. Individual and median group responses are shown. Data include all animals from previous studies vaccinated identically with RH5, CyRPA or RIPR at the indicated dose. Significance determined by one-way ANOVA with Dunnett’s multiple comparisons test versus RH5 only group, * p < 0.0104, ** p < 0.0017, *** p < 0.0005, **** p < 0.0001. RH5: n = 15, CyRPA: n = 12, RIPR: n = 11, all other groups n = 6 biologically independent experiments. H GIA EC50 data from the indicated groups (from data in Supplementary Fig. 8C). The Dotted line indicates the median result for RH5-only immunised animals (2 µg dose) for comparison. RH5: n = 21, CyRPA: n = 12, RIPR: n = 11, RCR: n = 24, and R78C + RH5 n = 12 biologically independent experiments. I Combined day 70 serum IgG ELISA data (summed in µg/mL) for all three antigens across select vaccination groups receiving the same immunogens; 2 µg dose RH5 data is shown. The dashed line indicates the median result for RH5-only immunised animals for comparison. RH5: n = 15, CyRPA: n = 12, RIPR: n = 11, all other groups n = 6 biologically independent experiments. J Summary of data shown in (I) with the median contribution of each different antigen-specific IgG (in µg/mL) displayed for each immunisation group. RH5 (2 µg): red; CyRPA: blue; RIPR: green. The dashed line shows the median result for the RH5-only group for comparison. Source data are provided as a Source Data file.