NSG Mouse Model for AML Therapy

All animal studies were carried out under protocols approved by the Johns Hopkins Institutional Animal Care and Use Committee. NSG (NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ) mice 6–8 weeks old were obtained from an internal colony that originated from the Jackson Laboratory (Bar Harbor, Maine). Mice were injected with 1×10⁶ MV-4-11 cells modified for stable firefly luciferase (ffLuc) expression31 (link) or 5×10⁴ MOLM-13.ffLuc cells32 (link) via tail vein on D0. NK-cell treatment was administered on D7 (10×10⁶ cells) or D4, 7, and 10 (3×10⁶ cells each). Mice were given D-luciferin (3 mg) by intraperitoneal injection, and bioluminescence (BL) was measured using IVIS Spectrum (In Vivo Imaging System). Data were analyzed using Living Image Software V.4.7.3 (PerkinElmer, Waltham, Massachusetts, USA). When indicated, peripheral blood (PB) was drawn via facial vein; red blood cells were lysed with eBioscience RBC Lysis Buffer (Thermo Fisher); and the remaining cells were analyzed with flow cytometry. Bone marrow and spleen were harvested, and tissues were analyzed with flow cytometry. Analysis of PB for cytokines (hIL-15, hTumor Necrosis Factor (TNF)-α, mouse interleukin (mIL)-6, and mIL-1β) was performed with ELISA (R&D Systems). Mice were euthanized when they exhibited >20% weight loss, hind limb paralysis, or moribund state as per protocol guidelines.