Humanized mouse model

Humanized mice (HIS mice) were generated in our laboratory. Briefly, immune-deficient NSG (NOD.Cg-Prkdc_scidIl2rg_tm1Wjl/SzJ) mice were injected intrahepatically with 100,000 human CD34⁺ hematopoietic stem cells (HSC) and analysed for human immune cell reconstitution after 12 weeks using multi-colour antibody panels established for immune-phenotyping of human whole blood samples as described⁴⁷, with some minor modifications to include ILC reconstitution (Lin⁻CD127⁺ cells). Once reconstituted, 12–14 weeks post transplantation, HIS mice, either males or females, were i.v. injected with enhanced luciferase⁴⁸ transfected HL60 APL cells previously tested for mycoplasma contamination (2.5 × 10⁶ live cells/mouse). ATRA (Sigma) (65 mg/kg body weight, i.p.) was administered daily from day 21 to 24. Hundred and fifty μg/mouse/dose of anti-NKp30 and anti-IL-13 were injected every 5 days; to monitor leukaemia progression and treatment effectiveness, mice were injected with luciferin (3 mg/mouse) at days 7, 17, 25. Images were taken in the Xenogen Ivis 100 in vivo Imaging System 20 min after the injection (60 s, Bin4, F/Stop 1,2). Animals were randomly assigned to specific treatment groups, and the investigator was blinded to the group allocation. Human CD34⁺ cells were purchased from either Lonza (CH) or AllCells (USA). NSG mice were purchased from The Jackson Laboratory (stock number: 005557) and bred, treated and maintained under pathogen-free conditions in-house. Animal experimentation followed protocols approved by the local Veterinary Authority (VD Service de la consummation et des affaires Vétérinaires) under the licence VD2797 and performed in accordance with Swiss ethical guidelines.