Fig 9. Low IFNγ transcriptional CD8 T cell responses to T. gondii-infected Nlrp3-/- BMDMs are not due to dysregulated co-stimulatory pathways.

(A-B) Wildtype (WT) and Nlrp3-/- BMDMs were infected with clade D ME49 or clade B MAS strains. T-GREAT CD8 T cell responses to parasite-infected BMDMs were analyzed as described in Fig 6A. (A) The frequency of activated CD8 T cells (CD69+ CD62L- CD8+ T cells), and (B) the frequency of YFP+ (Ifng:YFP+) cells among activated CD69+ CD62L- CD8 T cells were determined. Representative flow plots with indicated gates and frequencies are shown from 3–4 experiments. (C-D) WT and Nlrp3-/- BMDMs were infected with a GFP-expressing Pru strain (Pru A7) and stained for the indicated co -stimulatory and -inhibitory molecules at 18h. (C) Surface marker expression of infected (GFP+) and uninfected (GFP-) WT BMDMs from an infected well, and of BMDMs from uninfected control wells are all compared in a single histogram plot; the isotype staining control is also indicated. A representative histogram plot from 2–3 independent experiments is shown for each marker. The gating strategy is depicted in S5A Fig. (D) As in C, but infected WT and Nlrp3-/- BMDMs (GFP+) are compared; isotype staining control of infected cells is also indicated. A representative histogram plot of 2–3 independent experiments is shown for each marker.