Figure 3. Functional characterization of H2-D^k transgenic mice.

(A) Schematic representation of the H2-D^k gene within the 11.5 kb EcoRI genomic DNA fragment from AKR mice used to generate transgenic mice. (B) Expression of H2-D^k on MEFs from FVB/N nontransgenic (FVB-Tg(D^k)⁻, transgenic FVB-Tg(D^k)⁺, and AKR (H2^k) mice. MEFs from FVB-Tg(D^k)⁺ and wild-type littermates were prepared from PCR-typed embryos, untreated or incubated overnight with 100 U/ml IFN-β prior to analysis of H2-D^k expression by FACS. (C) H2-D^k staining on lymphocytes from FVB-Tg(D^k)⁻ (red peak), BALB.K (dashed peak), MA/My (black peak), and FVB-Tg(D^k)⁺ (dotted peak) mice. Bar graph shows quantification of the percentage of H2-D^k expression from 2–3 mice per group. (D) Splenocytes from B6.H2⁰ mice or NKC/H2 histocompatible mice were inoculated into either untreated or asialo-GM1–treated NK cell–depleted hosts. Ratio values indicate the relative survival in the test population (CFSE^high) compared to the histocompatible control population (CFSE^low) at 18 hours after injection. Three mice per group were analyzed. Statistical significance between untreated and NK-depleted mice is shown. (E) Viral load in spleens (left) and livers (right) of mice of the indicated genotypes was determined by plaque-forming assays at day 3 p.i. Data were analyzed using two-way ANOVA analysis and the two-tailed Student's test. Data are presented as mean ± SEM and P values of significant results between groups are indicated. Results shown are representative of 2–3 independent experiments.