Figure 6.

Induced Eomes Deletion Does Not Significantly Alter NK Cell Functional Responses but Impairs Ex Vivo Cytotoxicity

(A and B) ILC-Eomes^Δ/Δ or Eomes WT mice were treated with the Tam-3d regimen. On day 2 of tamoxifen, mice were also administered 300 μg poly(I:C) intraperitoneally (i.p.) 24 h later, NK cells were enriched from splenocytes pooled from 2-3 mice per group and used in a flow-based killing assay against RMA-S targets. (A) Percent specific killing of RMA-S cells at the indicated effector/target ratios and (B) mean ± SEM. NK cell percentage of CD45⁺ cells from splenocytes used in the killing assay. n = 3 sets of pooled mice per group, three independent experiments.

(C) Mean ± SEM granzyme B percent positive and MFI of NK cells immediately before (in the blood) and 24 h after (in the spleen) poly(I:C). n = 7–11 mice per group, two or three independent experiments.

(D and E) Splenocytes from ILC-Eomes^Δ/Δ or Eomes WT mice treated with the Tam-3d, Tam-6d, or Tam-9d regimens were stimulated with plate-bound anti-NK1.1, IL-12 (10 ng/mL) + IL-15 (10 ng/mL) or YAC-1 lymphoma targets (10:1 effector:target [E:T]) in a 6-h functional assay. Summary data show (D) overall mean ± SEM percent IFN-γ or CD107a positive NK cells and (E) IFN-γ and CD107a positivity by stage in response to the different stimuli. n = 8–9 mice per group, three independent experiments. Data were compared using false-discovery-rate-corrected t tests.

*p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.