Figure 7. Th-POK is necessary for the normal functional response of Vα14i NKT cells. (A) Reduced CD69 expression. Histograms depicting the expression of CD44, NK1.1, and CD69 on Vα14i NKT cells from the livers and spleens of hd/hd (red) and +/+ (blue) littermates, with the staining of WT CD8 SP T cells (green) overlain for comparison. Results are representative of at least six independent experiments. (B) Reduced IL-4 production. Intracellular cytokine staining of Vα14i NKT cells from hd/hd and WT mice two h after injection of αGalCer or vehicle. Liver and spleen cell suspensions were stained with αGalCer-CD1d tetramers and anti–TCR-β, after which cells were fixed, permeabilized, and stained with antibodies against IL-4 and IFN-γ. Data are representative of six independent experiments. (C) ELISA was used to measure IL-4 in blood drawn from the indicated strains of mice injected 2 h previously with αGalCer. Similar results were obtained in separate assays of two other sets of hd/hd and +/+ or hd/+ littermates challenged with αGalCer (not depicted). (D) The functional effect of the hd mutation in Vα14i NKT cells is independent of CD8 expression. Staining for IL-4, IFN-γ, and TNF in permeabilized liver Vα14i NKT cells from mice with the indicated genotypes heterozygous or homozygous for the hd mutation and a disrupted CD8α allele. Cells were harvested 90 min after αGalCer challenge. Similar responses were observed in a second pair of hd/hd, CD8α−/−, and hd/hd CD8α+/− littermates.
Figure 7.

Th-POK is necessary for the normal functional response of Vα14i NKT cells. (A) Reduced CD69 expression. Histograms depicting the expression of CD44, NK1.1, and CD69 on Vα14i NKT cells from the livers and spleens of hd/hd (red) and +/+ (blue) littermates, with the staining of WT CD8 SP T cells (green) overlain for comparison. Results are representative of at least six independent experiments. (B) Reduced IL-4 production. Intracellular cytokine staining of Vα14i NKT cells from hd/hd and WT mice two h after injection of αGalCer or vehicle. Liver and spleen cell suspensions were stained with αGalCer-CD1d tetramers and anti–TCR-β, after which cells were fixed, permeabilized, and stained with antibodies against IL-4 and IFN-γ. Data are representative of six independent experiments. (C) ELISA was used to measure IL-4 in blood drawn from the indicated strains of mice injected 2 h previously with αGalCer. Similar results were obtained in separate assays of two other sets of hd/hd and +/+ or hd/+ littermates challenged with αGalCer (not depicted). (D) The functional effect of the hd mutation in Vα14i NKT cells is independent of CD8 expression. Staining for IL-4, IFN-γ, and TNF in permeabilized liver Vα14i NKT cells from mice with the indicated genotypes heterozygous or homozygous for the hd mutation and a disrupted CD8α allele. Cells were harvested 90 min after αGalCer challenge. Similar responses were observed in a second pair of hd/hd, CD8α−/−, and hd/hd CD8α+/− littermates.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal