Fig. 2

Defective killing activity of NFATc1-deficient aCD8⁺ Ts and CTLs. a Defective killing of MOPC 315 plasmacytoma cells expressing a luciferase indicator gene by aCD8⁺ Ts generated upon activation by αCD3/CD28 for 3 d in vitro. WT, Nfatc1 ^−/− (Nfatc1 ^f/f x CD4-cre mice) and Nfatc2 ^−/− aCD8⁺ Ts were incubated with MOPC target cells for 24 h. As controls, naive CD8⁺ T cells were incubated, and WT aCD8⁺ Ts were incubated without MOPC target cells, and WT aCD8⁺ Ts were incubated without MOPC target cells. b Defective killing of A20J target cells by Nfatc1 ^−/− CTLs. WT and Nfatc1 ^−/− C57/B6 CTLs were generated by incubation with irradiated splenocytes from Balb/c mice for 6 d, followed incubation with A20J Balb/c tumor cells for 2 h (see Supplementary Fig. 3b for details). Left, apoptosis induction of A20J cells is presented. Right, the degranulation of CTLs is shown. c Changes in number of NFATc1-deficient CTLs expressing various cytokines, granzyme B (Gzmb) or perforin (Prf1). Intracellular cytokine expression of CTLs was measured by flow cytometry upon stimulation by T + I for 6 h. Granzyme B and perforin expression was measured without T + I stimulation. d Defective clearance of Listeria monocytogenes upon infection in mice bearing NFATc1-deficient T cells. WT and Nfatc1 ^f/f x CD4-cre mice were infected with 5 × 10⁵ CFU ΔActA Lm-Ova⁶⁶ bacteria. 5 d later, the mice were sacrificed and Lm titers in livers were determined. e Distribution of splenic CTLs upon Lm infection of WT mice and Nfatc1 ^f/f x CD4-cre mice. Control, uninfected mice. f Number of cytokine-producing CTLs upon Lm infection in Nfatc1 ^f/f x CD4-cre mice. Data are shown as means ± SEM. Unpaired Student’s t-test was used for statistics