Fig. 5: IFNγ production by ex vivo stimulated PBMCs, splenocytes, and lung leukocytes following GP-Cda1/Cda2 vaccination and/or infection.

BALB/c mice were vaccinated thrice at biweekly intervals with GP-Cda1/Cda2. Two weeks after the last boost, the mice received a pulmonary challenge with C. neoformans. Mice were euthanized at 0 dpi (uninfected), 10 dpi, or 70 dpi. PBMCs (a), spleens (b), and lungs (c) were collected. Controls included unvaccinated mice that were euthanized at 0 dpi or 10 dpi. Single-cell PBMC, spleen, and lung suspensions were prepared following which cells were cultured in complete media supplemented with amphotericin B in the presence of the indicated antigens for either 3 days (PBMC and spleen) or 18 h (lung). Control cells were left unstimulated (Unstim). Supernatants were collected and analyzed for IFNγ by ELISA. Each group had five mice. Data were presented as mean ± SEM. Vac vaccinated with GP-Cda1/Cda2. Chall challenged with C. neoformans strain KN99. Dpi days post infection, HK KN99 heat-killed C. neoformans strain KN99. IFNγ production following SEB stimulation, as a positive control, was shown in Supplementary Fig. 2. The results of the statistical comparisons between groups were shown in Supplementary Fig. 3.