Figure 6.

Increased missing self-responses after αGC-treatment. Mice were injected i.v. with PBS, or αGC. 3 to 14 days after in vivo stimulation, survival of target cells was tested by challenging the mice with a mix of CFSE-labeled B6 and β₂m^−/− spleen cells. Remaining target cells were measured by flow cytometry. The survival ratio of remaining MHC-I⁻ (β₂m^−/−) target cells is shown, divided by remaining MHC-I⁺ (B6) control cells (relative survival). (A) Rejection capacity of NK cells in B6 mice was tested by injection of a mix of B6 and β₂m^−/− spleen cells at d3, d7, d10 and d14 after in vivo treatment with αGC. (B) Missing self-responses of C57Bl/6, MHC-I^−/−, CD1d^−/− or Jα18 were assessed at day 7 after in vivo αGC treatment. (C) B6 mice were treated with 200 ng or 1 µg of αGC (stimulation 1) and re-treated after 2 weeks (stimulation 2). Three days after the second treatment, the rejection capacity towards MHC-I-deficient spleen cells was assessed. Data are pooled from 2 independent experiments with 6 mice per group (A), 3 independent experiments with 9–12 mice per group for B6 and 4–6 mice per group for CD1d^−/−, Jα18^−/− and MHC-I^−/− mice (B), and 2 independent experiments with 5 mice per group (C). One-way ANOVA comparing each group to PBS-treated control group with Dunnett’s correction for multiple comparisons (A) or Tukey‘s multiple comparison test (C) or unpaired, two-tailed t-test (B) was used to determine statistical significance. Error bars denote SD. Significant differences are denoted * p < 0.05, ** p < 0.01, *** p < 0.001.