Nod ltsz scid il2rγnull mice

NSG (NOD/LtSz-SCID IL-2Rγnull) mice were obtained from the Jackson Laboratory and were housed at the Veterinary Service Center of Stanford University. The use of animals in the study adhered to the guidelines and regulations established by the Institutional Animal Care and Use Committee (IACUC) of Stanford University. PC3 control vector, PC3 AZPG1-OV, DU145 control and DU145 AZGP1-OV (1 × 10⁶ cells), 22Rv1 control and 22Rv1 AZGP1-OV (2 × 10⁶ cells) were suspended in 50% Matrigel (100 µL) and were injected subcutaneously into the dorsal flank of 10–12 week-old NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ (NSG) mice (The Jackson Laboratory, Sacramento, CA, USA). The tumor size was measured every 3–4 days using a Vernier caliper, and tumor volume was calculated as V = 1/2 × L × W². At the end of experiment, mice were sacrificed, and the tumors were collected. Half of the tissues were formalin-fixed and paraffin-embedded and the other half snap frozen for proteomic analysis by mass spectrometry.

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.