Fig. 2
Optimisation of the ex vivo MGIA using BCG Pasteur Aeras. a 1 × 106, 3 × 106, and 5 × 106 splenocytes (1 M, 3 M, and 5 M, respectively) from mice immunized with BCG Pasteur Aeras (grey circles) or given saline (open squares) were co-cultured with 2000 CFU (a), 500 CFU (b) or 100 CFU (c) of BCG Pasteur Aeras. Splenocytes were obtained from a total of 6 immunized and 6 control animals. Aliquots from each spleen were cultured with different numbers of mycobacteria as indicated, and are represented by individual data points. Error bars represent the median +/- interquartile range. Statistical significance was tested using the unpaired t test function in GraphPad Prism. d Analysis of variation of the data shown in a-c. *Sample sizes (alpha 0.05, power 0.8) were calculated using the mean values of BCG immunized and naïve groups and using the standard deviation of the BCG group (standard deviation was greater in the BCG groups than in the saline groups). **Optimal conditions for vaccine candidate testing determined as delta >0.5 log CFU, CoV < 50 % and required sample size < 10 per group. The analysis was carried out using STATA software