Fig. 1: CD4 T cells infiltration to the C. neoformans infected brain correlates with fungal burden and accumulate near sites of yeast growth.

a Fungal burdens in the brain at days 0, 3, 6 and 9 post-infection. Data from a single experiment, n = 3 mice (day 3 and 6) or 4 mice (day 9). Data presented as mean +/- SEM. b Weight loss, relative to starting weight, following i.v. infection with C. neoformans H99 in C57BL/6 mice. Data pooled from two independent experiments, n = 11 mice per time point. Data show as mean +/- SEM. c Total number of CD4 T cells per brain at days 0 (n = 5 mice), 3 (n = 4 mice), 6 (n = 7 mice) and 9 (n = 6 mice) post-infection. Data pooled from 1 to 2 independent experiments. Whiskers refer to the maximum/minimum values, the box refers to interquartile ranges, the centre line refers to the mean. d Number of brain-infiltrating CD4 T cells relative to brain fungal burden in mice infected with low (open symbol), medium (grey symbol) and high (filled symbol) dose of C. neoformans. Each point represents an individual animal. Data analysed by simple linear regression. e Example flow cytometry plots of peripheral blood and brain samples (matched from same animal) from mice pre-injected with cell labelling dye CFSE 10 min prior to euthanasia and analysis. Graph shows quantification where each point represents an individual animal. Data analysed by paired t-test. *P = 0.0206. f Example confocal microscopy images of CD4 staining in the infected brain at day 7 post-infection (representative of 3 animals analysed). Wild-type mice were infected with GFP-expressing C. neoformans and brain sections stained with DAPI (blue) and anti-CD4 (white). g Example Cda2-MHCII tetramer staining of leukocytes isolated from uninfected (open symbol) or C. neoformans-infected brains (at day 7 post-infection; closed symbol). Example dot plots are pre-gated on CD45^hiCD44⁺CD4⁺TCRβ⁺ live singlets. Frequency and the number of cells acquired were quantified in uninfected (n = 5 mice) and infected (n = 11 mice) animals. Data pooled from two independent experiments and analysed by Mann-Whitney U-tests. *P = 0.0389, **P < 0.0094.