Fig. 8. Type I interferons enhance survival of preferentially Ly6C⁺ CD8⁺ Tn cells in vitro.

A We magnetically enriched CD8⁺ Tn cells to >95% purity. 3 × 10⁵ cells were cultured in round bottom 96 well plates either alone or in the presence CD45- LN stroma in RPMI media + 10% FCS and 0.05 mM beta-mercaptoethanol ±100 in ng/ml IFN-α. Cell viability was assessed by flow cytometry on days 1, 3, 5, and 7 of culture (n = 8 mice, data presented as mean ± sd). B viability of wt CD8⁺ Tn cells cocultured with Ifnar1^−/− stroma and vice versa Ifnar1^−/− CD8⁺ Tn cells stimulated by 100 ng/ml IFN-α was measured on days 1, 3, 5, and 7 (n = 8 mice, data presented as mean ± sd). C expression of Ly6C on live CD8⁺ Tn wt cells cultured with LN stroma with or without IFN-α (n = 8 mice, data presented as mean ± sd). Data are pooled from two experiments, n = 8, (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). A, B, C For the survival of cultured T cells cultured with or without stroma, statistical significance was calculated with a paired two-tailed Student’s t-test between corresponding time points.