Establishment of Humanized Mouse Model

Rag2^−/−γc^−/− mice were purchased from Taconic and maintained in the Laboratory Animal Unit, the University of Hong Kong. To establish humanized mice models, huPBMCs were isolated from buffy coat preparations of blood from healthy donors from the Hong Kong Red Cross. Vγ9Vδ2 T cell–depleted huPBMCs were obtained after depletion of Vδ2 T cells by magnetic microbeads (Miltenyi Biotec). 4–5-wk-old male or female Rag2^−/−γc^−/− mice were treated with liposomes (VU Medisch Centrum, Amsterdam, the Netherlands) 1 d before transplantation. The sublethally irradiated mice were transplanted i.p. with 30 × 10⁶ huPBMCs or Vγ9Vδ2 T cell–depleted huPBMCs. In general, huPBMCs from 1 buffy coat were used for generation of 8–10 humanized mice. After transplantation, graft-versus-host disease symptoms such as weight loss, temperature changes, and diarrhea were monitored daily. The immune reconstitution of humanized mice was examined at indicated times. All manipulations were performed in compliance with the approval of the Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster and the guidelines for the use of experimental animals by the Committee on the Use of Live Animals in Teaching and Research, Hong Kong.