Nod cg prkdcscid il2rgtm1wjl szj mice

BALB/c Nude mice CAnN.Cg-Foxn1nu/Crl (Charles River) were injected with 6 × 10⁶ A375 melanoma cells in the subcutaneous dorsal area. Approximately one week later, the tumor reached a volume of about 100 mm³, at which point mice were assigned to four tumor size-comparable groups of 12 animals and treated as described in the Supplementary Methods.
Fresh tissue from patient 17, who had been diagnosed with metastatic melanoma (Table I and Figure 5), was minced and xeno-injected into NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ mice (commonly known as NOD scid gamma (NSG) mice) (Charles River). Briefly, the animals were anesthetized using ketamine (75 mg/kg) and medetomidine (1.0 mg/kg) and a piece of tumor was inserted in the subcutaneous dorsal area through a small incision in the skin and allowed to grow. Next, mice were sacrificed (as described in supplementary methods) and tumors were collected and minced into pieces of about 2 mm³ and reimplanted into the experimental group of mice. When these mice had grown tumors of an approximate volume of 100 mm³, they were distributed among four groups of six mice, each with comparable tumor volumes and treatments were started, as described in Figure 5 and in the Supplementary Methods.

NOD.cg-Prkdc^SCID Il2rg^tm1Wjl/Szj mice (abbreviated as NOD-scid IL-2Rγ^null) were obtained from Charles River Laboratories (Sulzfeld, Germany). Mice were kept under specific pathogen-free conditions in individually ventilated cages. The facility was controlled according to the Federation of Laboratory Animal Science Association (FELASA) guidelines. Following engraftment (day 1), mice were pre-sensitized through rectal application of 150 µl of 10% ethanol on day 8 using a 1-mm cat catheter (Henry Schein, Hamburg, Germany). The catheter was lubricated with Xylocain^© Gel 2% (AstraZeneca, Wedel). Rectal application was performed under general anesthesia using 4% isoflurane. Post application, mice were kept at an angle of 30° to avoid ethanol dripping. On days 15 and 18, mice were challenged with rectal application of 50% ethanol following the protocol described for day 8. Mice were killed on day 21. Pitrakinra (10 µg in 0.5% methylcellulose, 0.05% Tween-80) in PBS (Zadeh-Khorasani et al., 2013 (link)) was applied on days 7-9 and 14-21. In these groups, sterile saline (B. Braun Melsungen AG, Germany) served as control. Infliximab [6 mg/kg (Remicade^©, Janssen, The Netherlands)] was applied on days 7, 14 and 17. An isotype antibody (human IgG1, kindly provided by, MorphoSys AG) was used as control. All treatments were applied intraperitoneally.