Fig. 4

Old mice generate more TNFα-producing G6PI-specific Th cells. a–d Peripheral LN cells from young (average age of 8 weeks, n=5) or old (average age of 103 weeks, n=5) mice were restimulated with anti-CD3/CD28 beads (0.5 beads/cell) for 6h, fixed and intracellularly stained for IFNγ, IL-4, IL-17A, GM-CSF, and TNFα in combination with CD154 and CD4. FACS plots show representative staining for CD154⁺ among CD4⁺ cells (a) and for cytokine producers among CD154⁺CD4⁺ cells (b). Diagrams show average frequencies of CD154⁺ cells among CD4⁺ cells (c) or of cytokine producers among CD154⁺CD4⁺ cells (d). e–g Peripheral LNs were collected from young (average age of 9 weeks, n=7) and old mice (average age of 89 weeks, n=7) at day 6 after immunization with G6PI. Whole LN cells were restimulated with G6PI for 6h, fixed and stained intracellularly for cytokines in combination with CD154 and CD4. FACS plots show representative staining for CD154⁺ among CD4⁺ cells (e). Diagrams show average frequencies of CD154⁺ among CD4⁺ cells (f) or of indicated cytokines among CD154⁺CD4⁺ (g). Statistical testing was performed as described in the “Methods” section. n.s. non-significant