Fig. 6: Protective effect of CTH522/CAF®01 induced immune responses.

a Experimental outline. B6C3F1 mice were immunized with 10 µg CTH522/CAF®01 or CAF®01 alone. b CTH522 specific T cell responses. Splenocytes (n = 6) were stimulated in vitro with CTH522 and IFN-γ and IL-17A responses were measured in supernatants. Data points represent cytokine levels in antigen stimulated wells after subtraction of media only for each mouse and each line represents median level with 25th and 75th percentiles. Two-tailed Mann–Whitney U test was used for comparison among groups. c Serum from vaccinated mice (n = 63) and control serum samples (n = 6) were isolated 4 weeks post last vaccination and CTH522 specific IgG analyzed by ELISA. Data points represent the median OD value with 25th and 75th percentiles at each titration step. d Vaginal wash samples were collected from CTH522 vaccinated mice (n = 63) and control mice (n = 11), diluted 5 times, and CTH522 specific IgG were measured. Results are presented as a Box and Whiskers plot with median and 25th and 75th percentiles (Box) and Min to Max, all data points (Whiskers). Two-tailed Mann–Whitney U test was used for comparison among groups. e Mice were challenged i.vag. with 1 × 10⁵ IFU/mouse of C.t. SvD. IFU were recovered from vaginal swabs at day 3, 7, 10 (n = 63), 14, and 17 (n = 31) post infection. Data are presented as a Box and Whiskers plots with median and 25th and 75th percentiles (Box) and Min to Max, all data points (Whiskers). Two-tailed Mann–Whitney U test was used for comparison among groups. f Area under the curve (AUC) from PID3 to PID17 was calculated for the 31 mice proceeding to PID17 and results presented as a Box and Whiskers plot with median and 25th and 75th percentiles (Box) and Min to Max, all data points (Whiskers). Two tailed Mann–Whitney U test was used for comparison among groups. The experiment has been repeated three times with similar results. g Protection against ascending infection divided into mGT; Uterine horns and uGT; Oviducts/ovaries. The numbers of culture positive samples/total number in the individual experiment as well as a sum of the three experiments are presented. Sum of mGT C.t. positive vs. negative in the two groups, Two-sided Fisher’s exact test, p < 0.0001. Sum of uGT C.t. positive vs. negative in the two groups, Two-sided Fisher’s exact test, p = 0.0006. a Figure was created with Biorender.com. b–f Source data are provided as a Source Data file.