Cb117

Commercially purchased CD34-depleted human cord blood mononuclear cells (AllCells, LLC., CB117) were infected with M81 strain virus using 2000 infectious units. Cord blood was initially exposed to the virus in vitro for 1.5 hours and then 10 to 25 million cells were injected intraperitoneally (i.p.) into 3–5 week old NSG mice. Mice were treated with 20 mg/kg teriflunomide starting on day 4, three times a week. Mice were euthanized on day 35 (Experiment 1) or 28 (Experiment 2). Tumor size was quantitated by dissecting and weighing grossly visible tumor tissue.

Immunodeficient nonobese diabetic/severe combined immunodeficient (NOD/LtSz-scid/IL2Rγ^null) mice were purchased from Jackson Labs (catalogue 005557). Commercially purchased CD34-depleted human cord blood mononuclear cells (AllCells, LLC., CB117) were mock-infected, or infected with M81 strain EBV, in vitro for 1.5 hours, and then 12 to 25 million cells were injected intraperitoneally (i.p.) into 3–5 week old NSG mice. Each individual treatment experiment (with or without blocking antibodies) was performed in animals injected with the same number of cord blood cells (all from the same donor), and the same amount of infectious M81 virus.

EBV⁺ B-cell lymphomas were generated in immunodeficient NSG (NOD/LtSz-scid/IL2Rγ^null) mice (Jackson Labs) as previously described [46 (link)]. In brief, CD34-depleted human cord blood mononuclear cells (#CB117; AllCells) were infected in vitro with the M81 strain of EBV (2,000 GRU) by incubation at 37°C for 1.5 h after which the infected cells were injected i.p. into 3- to 5-week-old NSG mice. Thirty-three days later, the mice were injected i.p. with 60 mg/kg of Hypoxyprobe (Hypoxyprobe) and sacrificed 1.5 h later by cervical dislocation under isoflurane anesthesia. Portions of the harvested tumors, along with some internal organs as controls, were submerged in Optimal Cutting Temperature compound and flash-frozen in ethanol-dry ice. Other portions of tumors, along with internal organs, were formalin-fixed and paraffin-embedded for sectioning and mounted onto slides for IHC.
EBV⁺ gastric cancer xenografts were generated by subcutaneous inoculation of 1x10⁷ SNU-719 cells in Matrigel into the flanks of NSG mice. Thirty-three days later, the mice were injected i.p. with 60 mg/kg of Hypoxyprobe and sacrificed 1.5 h later. Portions of the tumors were prepared as described above.