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. 2022 Sep 9;13:953151. doi: 10.3389/fimmu.2022.953151

Figure 6.

Figure 6

iNKT cells restrain antiviral effector functions of virus-specific CD8+ T cells. Two immune-dominant MHC class I restricted NrHV epitopes were identified using an overlapping NrHV peptide library. (A) Representative FACS plots showing intracellular IFN-γ production by hepatic CD8+ T cell isolated at day 21 pi and stimulated with 10ug/ml of the NrHV NS3 or NS4 epitope for 5h. Gated on CD8+ T cells. (B) Summarized data of (A). (C) IFN-γ production by CD8+ T cells in response to stimulation with increasing concentrations of the NS3 or NS4 peptide. (D) FACS plots showing hepatic NS3- or NS4- specific CD8+ T cells at day 21 pi using NS3- or NS4- MHC class I tetramer staining. Gated on CD3+ T cells. (E) Frequencies of NS3- and NS4-specific cells within the hepatic CD8+ T cell population in WT and CD1d-/- mice (n=4 per group) at day 14 pi (left graph) and CD69 and T-bet expression on NS3- and NS4-specific CD8+ T cells (right graphs) at day 14 pi. (F-G) CD8+ T cells were isolated at day 14 pi from WT and CD1d-/- mice (n=4 per group), stimulated with the NS3 or NS4 peptide or left unstimulated and analyzed for antigen-specific granzyme B/perforin and IFN-γ expression. Summarized data (NS3 and NS4 combined) is shown in (F) and representative FACS plots in (G). Unpaired t test: *p< 0.5, **p<0.01. ns, not significant.