Figure 4.

iNKT Cell Expansion in Map3k1-Deficient Mice

(A) WT, Map3k1^ΔKD, and Lck^Cre/+Map3k1^f/f mice were i.p. injected with α-GalCer for 3 days. Splenocytes were harvested at days 0 and 3, stained with anti-TCRβ antibody and CD1d tetramer, and analyzed by flow cytometry as indicated. Data are representative of three independent experiments. Numbers in the profiles indicate the percentages of the gated populations.

(B) Statistical analysis of α-GalCer-dependent iNKT expansion at days 0, 3, and 6 in Map3k1^ΔKD mice. The average percentage (±SEM) of PBS-57-loaded CD1d tetramer⁺ TCRβ ⁺ cells from five independent experiments is shown (black circle, WT; black square, Map3k1^ΔKD mice). Differences were analyzed by two-tailed Student’s t test (^∗p ≤ 0.05; ^∗∗p ≤ 0.01; ^∗∗∗p ≤ 0.001).

(C) Liver cells were harvested at days 0 and 3 from WT, Map3k1^ΔKD, and Lck^Cre/+Map3k1^f/f mice following i.p. immunization with α-GalCer. Liver cells were stained with CD1d tetramer and anti-TCRβ antibodies and analyzed by flow cytometry as indicated. Data are representative of three independent experiments. Numbers in the profiles indicate the percentages of the gated populations.

(D) Representative H&E-stained liver sections were prepared from unstimulated (upper panels) and 3-day α-GalCer stimulated (lower panels) WT, Map3k1^ΔKD, and Lck^Cre/+Map3k1^f/f mice (original magnification ×40; scale bar, 10 μM). Arrows indicate lymphocyte infiltration. Data are representative of three independent experiments (two mice per experiment).