Evaluating Human KIR B Haplotype in Mice

KIR transgenic mouse expressing a complete human KIR B haplotype on a C57BL6 background MHC class I–deficient Kb^−/− Db^−/− were a kind gift from J Van Bergen and kept under specific pathogen-free conditions.35 (link) DNA constructs were made expressing HLA-C*0102 alone or linked with the T2A self-cleaving peptide and peptide and cloned into pIB2.18 (link) For the B16 model, mice were injected intramuscularly with 50 µg DNA; 2.5×10⁵ B16F10 cells into the mice left flank, and 50 µg of DNA plasmid on days 0 and 7. For the Huh7 model, NSG mice were injected with 2×10⁶ Huh7-C*0102 cells subcutaneously and then 1×10⁶ purified NK cells from 2 weeks vaccinated KIR transgenic (KIR-Tg) mice spleen were injected intravenously on days 0 and 14. NK cells were purified using MACS technology (NK Cell Isolation Kit II, Miltenyi Biotec). Purity was > 90% NK cells with <3% CD3+ T cells.