Figure 3
Dendritic cells are required for NK-cell effector functions in vivo and in vitro.
(a) Experimental scheme for CFSE-based in vivo rejection assay. (b) Kinetic of in-vivo survival of MHC-class I-deficient target cells. (c–e) In vitro activating receptor stimulation of NK cells from mice after 4 days of DT injection. (Filled circles) littermate controls, (Empty squares) CD11c.DOG. (c) Representative plots of IFN-γ and CD107a expression after anti-NKp46 stimulation. (d) Summary of IFN-γ and CD107a expression on NK cells after anti-NKp46 and anti-NK1.1 stimulation. (e) Summary of IFN-γ and CD107a expression on NK cells of different maturation stages upon anti-NKp46 stimulation. (f,g) Ca2+ mobilization upon activating receptor stimulation after 4 days of DT injection. (f) Representative plots showing the calcium flux response of NK cells after stimulation of the NK1.1 receptor using antibodies and crosslinker. The arrow indicates the time when the crosslinker was added. (g) Summary plots of 3 experiments where the area under the curve was determined using values before addition of the crosslinker as a baseline. (h–j) Purified NK cells at 4 days after the start of DT injection were stimulated with IL-12 (1 ng/ml) and IL-18 (1 ng/ml). (h) Representative plots of IFN-γ and CD107a expression on NK cells upon IL-12 plus IL-18 stimulation. (i,j) Summary of IFN-γ expression on NK cells upon IL-12 plus IL-18 stimulation. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 (unpaired Student’s t test). Data shown are representative of 2 independent experiments (b), combined from 2 independent experiments (d,e,i,j), combined of 3 independent experiments (g). Error bars, SEM (n = 3–4 mice per group [b], n = 6–7 mice per group [g], n = 7 mice per group [d,e,i,j]).
