Cytokine recall responses in macrophages from <i>C</i>. <i>neoformans</i> H99γ immunized mice require epigenetic modifications.

BALB/c mice received an initial immunization with heat killed C. neoformans strain H99γ or C. neoformans strain H99γ and allowed to rest for 60 days. Macrophages were then isolated from the spleens of the immunized mice, as well as from naïve mice, and cultured ex vivo with C. neoformans strain cna1Δ +/- MTA, pargyline hydrochloride, givinostat or EGCG for 24 h (a-c) or C. neoformans strain cna1Δ +/- GSK343, UNC 0638 or MI-2 for 24 h (d-g). Supernatants were collected and analyzed for IL-2 (a, d), IFN-γ (b, e) and TNF-α (c, f) production. Alternatively, macrophages were collected after the 24h period and analyzed for total and phosphorylated STAT1 (g). Data shown are means +/- SEM of 3 independent experiments using 20–25 mice per group. Protein expression is representative of 3 independent experiments. (*p cna1Δ within the same immunization group).