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. 2024 Feb 13;15:1337. doi: 10.1038/s41467-024-45573-9

Fig. 6. Loss of Tyk2 impairs the cytotoxic activity of CD8+ CTLs.

Fig. 6

a Protein expression levels of GZMB in the unstimulated 8.3 CD8+ T cells, ex vivo-activated 8.3 CD8+ T cells, or ex vivo-activated 8.3 CD8+ T cells cocultured with NIT-1 cells (Tyk2+/−, n = 6, Tyk2−/−, n = 6). Purified 8.3 CD8+ T cells were activated with IL-2, IL-12, and anti-CD3/28 beads for 3d. For coculture, ex vivo-activated 8.3 CD8+ T cells were cultured with NIT-1 cells for 24 h (CTLs:NIT-1 = 1:1). b Protein expression levels of GZMB in Tyk2+/− 8.3 CTLs after 24 h of coculture with NIT-1 cells in the presence of IFNAR1 blocking antibody (n = 7), IFN-β neutralizing antibody (n = 7), or IFN-α neutralizing antibody (n = 7). Purified Tyk2+/− 8.3 CD8+ T cells were activated with IL-2, IL-12, and anti-CD3/28 beads for 3d, and cultured with NIT-1 cells. c Gene expression levels of Ifnb1 in NIT-1 cells after coculture with ex vivo-activated 8.3 CD8+ T cells (0 h, n = 4; 1 h, n = 5; 2 h, n = 5; 6 h, n = 4; 12 h, n = 3). d The killing capacity of ex vivo-activated 8.3 CTLs against NIT-1 cells at the indicated ratios. Purified 8.3 CD8+ T cells were activated with IL-2 and CD3/28 beads in the presence or absence of IL-12 for 3d. NIT-1 cells were cultured with activated 8.3 CD8+ T cells for 24 h, and then the survival of NIT-1 cells was analyzed (1:0.1, n = 4; 1:1, n = 3; 1:2, n = 3). Data represent the mean ± SEM. P-values were calculated using one-way ANOVA with Dunnett’s posttest (c) and two-tailed Student’s t test (a, b, and d).