Fig. 3: Impaired memory formation of TNIK-deficient gp33-specific CD8⁺ T cells.

a Competitive AdTf (AdCoTf) model: 1 × 10⁵ splenic Tnik^WT (WT; Cd45.2⁺) and Tnik^−/− (KO; Cd45.1⁺) p14 T cells were AdCoTf into naive Cd45.1⁺Cd45.2⁺ mice previous to infection with 10⁴ pfu LCMV-WE. b Chimerism of splenic AdCoTf [(WT;black) vs (KO;red)] p14 T cells (n = 2–4 mice/group) 3, 7, or 10 days p.i. c Fold change of AnnexinV⁺ AdCoTf p14 T-cell frequency, normalized to average frequency of day 3 WT (n = 2–4 mice per timepoint). d Frequency of TNFα⁺, IFNγ⁺, and IL-2⁺ AdCoTf p14 T cells after in vitro re-stimulation with gp33 peptide (7 days p.i.). e, f ΔMFI of granzyme B, T-bet, and Eomes in WT/KO effector p14 T cells (n = 3 mice per group); normalized to isotype control. g Non-competitive AdTf (AdTf) model: 1 × 10⁵ splenic WT or KO (Cd45.1⁺) p14 T cells were AdTf into naive Cd45.2⁺ mice previous to infection with 10⁴ pfu LCMV-WE. h P14 T cell frequencies per total CD8⁺ T cells (blood) at indicated time points. i Numbers of AdTf p14 T cells (spleen) day 60 p.i. [representative data from two independent experiments (n = 5)]. j, k In total, 7 × 10⁵ CFSE-labeled memory p14 T cells (spleen; pooled from 5 to 7 primary recipients) were injected into congenic secondary recipients, followed by infection with 10⁴ pfu LCMV-WE i.v. j CFSE dilution of AdTf p14 T cells 3 days p.i.; non-activated CFSE^high (dotted line) p14 T cells (data from one out of two independent experiments). k Frequency and absolute numbers of AdTf p14 T cells in the spleen day 3 p.i. l Numbers of memory p14 T cells (blood/spleen) 40 days after AdTf of 10⁵ memory p14 T cells into secondary recipients and immunization (200 pfu LCMV; 1st boost). m Primary peak expansion rate (1st recipient) of KO p14 T cells normalized to WT T cells (n = 5 mice/group) and peak expansion rate of serially re-transferred and re-infected [(2nd recipient (n = 5 mice/group); 3rd recipient (n = 3 mice/group)] memory p14 T cells (blood). Depicted: WT p14 (black lines/circles), KO p14 (red lines/circles). Data are displayed as means ± SEM. Statistics: c, h two-way ANOVA, b, d–f, i, k–m two-tailed Student's t test, nonsignificant P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Also see Supplementary Fig. 3. Source data are provided as a Source Data file.