Fig. 5. T cell death and circulating chromatin release regulate neutrophil alterations and inflammation.

a Survival curves of WT, TCRα or RAG2-deficient mice infected with 5 × 10⁵C. albicans (n = 5, n = 8 and n = 4 mice per group respectively). b Colony forming units in the spleens and kidneys of WT and TCRα-deficient mice infected with 5 × 10⁵C. albicans, 72 h post infection. Representative of two experimental replicates with 8 mice per group. c Representative flow cytometric analysis of Ly6G^low and Ly6G^high neutrophils in the spleen of WT and TCRα-deficient mice infected with 5 × 10⁵ WT C. albicans and assessed as Live CD3⁻ CD19⁻ CD11b⁺ Ly6G^high/Ly6G^low 72 h post-infection. (WT n = 3 and TCRa-deficient n = 3 animals). Panel: Live/Dead blue, CD3/CD19 PerCP-Cy5.5, CD11b-PECy7, Ly6G-APC. d Ly6G^low/Ly6G^high neutrophil ratios in individual mice from (c). e Immunofluorescence micrographs from the spleens of naive or infected WT or TCRα-deficient mice infected with 5 × 10⁵ WT C. albicans, 72 h post-infection and stained for CD3 and poly-caspase enzyme activity performed with a FLICA poly-caspase activity assay (n = 3 animals per group and two independent experiments). Scale bars: 50 μm. f Immunofluorescence micrographs from the spleens of TCRα-deficient mice alone or after adoptive T cell transfer 48 h prior to infection and infected with 5 × 10⁵ WT C. albicans, 72 h post-infection (n = 3 mice per group and two independent experiments). Scale bar: 50 μm. g–i Cell-free DNA (g), western immunoblotting for histone H3 (h), and plasma cytokines and chemokines measured by multiplex immunoassay (i) of mice in f. Results from two independent experiments. Statistical analysis by unpaired two-sided Mann–Whitney t-test for single comparisons, two-tailed Log-rank (Mantel–Cox) test for survival analysis and two-way Anova for the cytokine array (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).