Fig. 2: Conditional deletion of Fgl2 from tumor-specific CD8⁺ T cells increases their persistence and improves tumor control.

a Schematic wherein Fgl2^−/− mice were given WT or Fgl2^−/− OCD8⁺ CD8⁺ T cells one day prior to cancer cell inoculation. Fourteen days later, immune cell function was measured via flow cytometry and bulk RNA-sequencing. b Tumor size as measured on day fourteen post tumor challenge, pooled data from three independent experiments, n = 15 mice in WT OT-I group and 13-15 mice in Fgl2^−/− OT-I group (p = 0.0253). c Representative flow plots and summary data showing d frequency and e cell count of CD8⁺Thy1.1⁺ cells present in the draining lymph node of mice given WT vs. Fgl2^−/− OT-I. Pooled data from two independent experiments, n = 10 mice per group, (p < 0.0001) and p = 0.0288, respectively). f Frequency and g cell count in the spleen of mice is also shown. Pooled data from two independent experiments, n = 9 in the WT OT-I group and 11 mice in the Fgl2^−/− OT-I group (p = 0.0097 and p = 0.0308, respectively). h Gene set enrichment analysis of differentially expressed genes found by bulk RNA-sequencing of WT vs Fgl2^−/− OT-I (isolated day 14 post tumor challenge) revealed that WTCD8⁺ D8⁺ D8⁺ T cells were significantly enriched for genes in the HALLMARK_APOPTOSIS MSigDB pathway compared Fgl2^-/− OT-I (n = 4 mice per group) with a normalized enrichment score (NES) = 1.332 and an adjusted p value = 0.0161. i Representative flow plots and j summary data showing the frequency of cleaved caspase 3/7⁺ WT and Fgl2^−/− OT-I, pooled data from two independent experiments (n = 9 in the WT OT-I group and 12 mice in the Fgl2^−/− OT-I group), (p = 0.0218). k Gene set enrichment analysis of differentially expressed genes revealed that Fgl2^-/− OT-I were significantly enriched for genes upregulated in memory compared to exhausted CD8⁺ T cells isolated day 30 post LCMV-Clone 13 infection compared WT OT-I (n = 4 mice per group) with a normalized enrichment score (NES) = 1.555 and an adjusted p value = 0.0003. l Representative flow plots and summary data in the m draining lymph node (n = 5 in the WT OT-I group and 6 mice in the Fgl2^−/− OT-I group) and n spleen (n = 4 in the WT OT-I group and 5 mice in the Fgl2^−/− OT-I group) showing frequency of PD-1⁺TIM-3⁺ OT⁻I within WT and Fgl2^−/− OT-I populations, (p = 0.0173 and p = 0.0159, respectively). Representative data from one of two independent repeats. o Schematic showing co-adoptive transfer wherein WT and Fgl2^−/− OT-I were transferred at a 1:1 ratio and mice were subsequently challenged with B16-OVA. p Representative flow plots gated on Thy1.1⁺ and summary data in theQ) dLN ad(R) tumor of WT (Thy1.1⁺) vs. Fgl2^-/− (Thy1.1⁺Thy1.2⁺) in tumor mice, n = 5 mice per group (p = 0.0159 and p = 0.0159, respectively). Mann-Whitney non-parametric, unpaired two-sided tests were used. The error bar in summary figures denotes mean ± SEM. *p < 0.05 **p < 0.01, ****p < 0.0001. Source data are provided as a Source Data file. a, o is created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.