αGC-loaded but not the unloaded CD1d-CD19 fusion promotes murine iNKT cell immunomodulatory functions in vivo. B6 mice were injected intraperitoneally with PBS44 (4 μg), αGC-loaded or unloaded CD1d-CD19 fusion (20 μg), or left untreated. (A) After 18 hours, splenocytes were analyzed for TCR⁺, CD4⁺CD8⁺, NK1.1⁺, B220⁺, and CD11c⁺ cells expressing CD69. Surface expression of the costimulatory molecule CD86 and MHC II on splenic CD19⁺ B cells (B) and CD11c⁺ myeloid cells (C) was assessed by flow cytometry. (D) Splenocytes were stained with NK1.1 and TCRβ antibodies and the NK1.1⁺TCRβ⁻ cells producing IFN-γ directly ex vivo were identified by intracellular staining and flow cytometry. Data are representative of 3 experiments with 1 to 2 mice analyzed per condition. Numbers in the histograms indicate MFI.