Proinflammatory networks are rapidly up-regulated in pulmonary macrophages of protectively immunized mice following challenge with <i>C</i>. <i>neoformans</i>.

BALB/c mice received an immunization with heat killed C. neoformans strain H99γ or C. neoformans strain H99γ and allowed to rest for 60 days. Mice were then challenged with WT C. neoformans strain H99 and subsequently sacrificed at day 1 or day 3 post-challenge and total RNA was extracted and sequenced from isolated pulmonary macrophages. Top canonical pathways and networks were predicted by the Ingenuity Pathway Analysis software. Top networks up-regulated in pulmonary macrophages from protectively immunized mice compared to non-protectively immunized mice as ranked by p-value (day 1, a; day 3, d). Blue and yellow represent genes up-regulated or down-regulated, respectively, in macrophages from protectively immunized compared to non-protectively immunized mice. Fold change values for up-regulated genes represented in networks (day 1, b; day 3, e). Top 5 immune response related gene ontology (GO) terms ranked by p value (day 1, c; day 3, f). Data were generated from a merged data set from 3 independent experiments with 20–25 mice per group.