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. 2014 Jul 25;124(9):3725–3740. doi: 10.1172/JCI72308

Figure 4. Cytokines activate αGalCer-pretreated iNKT cells.

Figure 4

(A and B) LPS (40 μg) was i.v. injected, as indicated, into C57BL/6 control (B6) mice or mice i.v. injected 1 month earlier with 4 μg αGalCer (B6/αGC) (3 mice/group). Animals were sacrificed after 6 hours, and splenic iNKT cells were analyzed for CD69 expression (P < 0.001 for B6 vs. B6/αGalCer; A), and IFN-γ expression was measured by intracellular cytokine staining (P = 0.345; B). Increase in CD69 expression based on geometric mean values. (C) αGalCer (1 μg) was i.v. injected into wild-type mice (B6), mice i.v. injected 6 weeks earlier with 4 μg αGalCer (B6 αGC), or mice i.v. injected 6 weeks earlier with 4 μg αGalCer and 2 weeks earlier with 40 μg i.v. LPS (B6/αGC + LPS) (3 mice/group). Splenic iNKT cells were analyzed 90 minutes later by intracellular cytokine staining for production of the indicated cytokines. P > 0.5 for B6/αGC versus B6/αGC/LPS for all 3 cytokines. Experimental outline is depicted in Supplemental Figure 2. Representative data from at least 2 independent experiments are shown in each panel.