NK cells negatively regulate CD8 T cells via natural cytotoxicity receptor (NCR) 1 during LCMV infection
Fig 2
Increase of virus-specific CD8 T cells in absence of NCR1 during acute LCMV infection.
(A) Frequency and total numbers of CD8 T cells in the spleen of naïve mice. Data shown are mean + SEM of n = 3 mice representative of 2 independent experiments. ns, not significant (unpaired two-tailed t test). (B) 1x103 P14 T cells (Ly5.1+) were transferred into WT and NCR1gfp/gfp mice followed by 200 ffu LCMV WE infection. After 7d, organs were harvested and flow cytometric analysis was performed. Number of CD8 T cells in indicated organs is shown. (C) 1x104 P14 T cells (Ly5.1+) were transferred into WT and NCR1gfp/gfp mice followed by 200 ffu LCMV docile infection. After 7d, organs were harvested and flow cytometric analysis was performed. Numbers of endogenous CD8 T cell subsets are shown. Data shown are mean + SEM of n = 5–6 mice pooled from 2 independent experiments. ns, not significant, * p<0.05 (unpaired two-tailed t-test). (D-J) 1x103 P14 T cells (Ly5.1+) were transferred into WT and NCR1gfp/gfp mice followed by 200 ffu LCMV WE infection. After 7d, organs were harvested and flow cytometric analysis was performed. (D) Representative flow cytometry plot pre-gated on CD8 T cells. (E) Representative flow cytometry plot pre-gated on endogenous CD8 T cells in the spleen. (F) Total number of transferred P14 (Ly5.1+), endogenous gp33-tet+ and endogenous np396-tet+ CD8 T cells in the spleen, (G) percentages and (H) total numbers in the lung. (I-J) Splenocytes were incubated in presence of Brefeldin A, gp33 and np396 peptide, respectively, for 6h at 37°C followed by intracellular cytokine staining. (I) Number of IFNγ and TNF producing CD8 T cells after gp33 stimulation is shown. (J) Number of IFNγ and TNF producing CD8 T cells after np396 stimulation is shown. Data shown are mean + SEM of n = 4 mice representative of 2 independent experiments. ns, not significant, * p<0.05,** p<0.01, *** p<0.001 (unpaired two-tailed t-test). (K) 1x103 P14 T cells (Ly5.1+) were transferred into WT and NCR1gfp/gfp mice followed by LCMV WE infection. After 5d, spleens were harvested and viral titers were determined. Data shown are mean ± SEM of n = 6–9 mice pooled from 2 independent experiments. ns, not significant, * p<0.05 (unpaired two-tailed t-test).