Fig. 5: Δbfd1 infected mice die of tachyzoite replication despite a competent immune response.

a, Survival of C57BL/6J mice infected with WT (n = 40 mice) or Δbfd1 (n = 40 mice) parasites. A subset of surviving mice received sulfadiazine treatment beginning at 21 d.p.i. (WT, n = 12 mice; Δbfd1, n = 15 mice; treatment start indicated by vertical dashed line; data representative of 3 independent experiments). b, Representative photomicrographs of WT-infected or Δbfd1-infected cerebra at 30 d.p.i. Insets show T. gondii cysts in WT-infected brains and tachyzoites in Δbfd1-infected brains (H&E; scale bars, 20 μm). c, Semiquantitative scoring of necrosis severity in brain sections at 30 d.p.i. (WT, n = 3 mice; Δbfd1, n = 4 mice; *P = 0.0286). d, Brain leukocyte cellularity during chronic infection with WT (n = 3 or 4 mice per time point) or ∆bfd1 (n = 3 or 5 mice per time point) parasites, quantified by Guava ViaCount assay (*P = 0.0142, ****P = <0.0001; data representative of 2 independent experiments). e–h, Flow cytometry analysis of the brains of naive mice (n = 4 mice) and mice at 30 d.p.i. with WT (n = 5 mice) or Δbfd1 (n = 3 mice) parasites (data representative of 3 independent experiments). e, UMAP and unsupervised x-shift clustering analysis of CNS CD45⁺ leukocytes at 30 d.p.i. f, Distribution of leukocytes among the 8 clusters shown in e (*P = 0.0001). g, Heat maps showing MFI of phenotypic markers with the highest expression in cluster 1. h, Quantification of inflammatory monocytes in the brain (CD45^highCD11b⁺F4/80⁺Ly6C^hi, cluster 1 shown in e; **P = 0.0085). Data analysed by two-sided Mann–Whitney test in c, two-way ANOVA in d, two-sided unpaired Student’s t-test with Bonferroni–Dunn correction for multiple comparisons in f and two-sided unpaired Student’s t-test in h. Bar graphs depict the mean ± s.d. Line graph depicts the mean ± s.e.m.

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