Foxn1nu nu

Female C57BL/6 mice were obtained from The Jackson Laboratory at 6–8 weeks of age Athymic nude mice Foxn1^nu/nu were obtained from Envigo at 4–6 weeks of age. All mice were maintained in a pathogen-free BSL2 biohazard facility. This facility is maintained between 68 and 79 °F with humidity from 30% to 70% and on a 12 h light/dark cycle. All animal studies were approved by the Institutional Animal Care and Use Committee at Mayo Clinic.

All animal experiments were performed using 8-week-old athymic nude mice (FoxN1^nu/nu, Envigo), following approval by the institutional animal care and use committee at SUNY Upstate Medical University. MPM cells were orthotopically implanted in the pleural space following a previously described model (42 (link)). Briefly, animals were anesthetized with isoflurane and cells were injected using a 0.5 ml insulin syringe (28G needle) in the right antero-lateral pleural space between the fourth and fifth ribs, 3-4 mm to the right of the sternum and to a depth of 4 mm. Cells (2x10⁶ H226 or 4x10⁶ H2595, expressing fLuc) were resuspended in 100 µL PBS. Tumor growth was monitored by bioluminescence and antibody treatment started when tumors had an integrated radiance signal above 1x10⁵ photons/sec x cm². Mice showing fLuc signal in the peritoneal cavity were considered mis-injected and discarded. mAb428.2 (30 mg/kg x day) or control human IgG were delivered by intrapleural injection for two weeks (5 days on + 2 days off). Tumors were monitored by bioluminescence for an additional month after treatment and mice were euthanized when they showed significant weight loss or additional symptoms of tumor burden. Due to the slow-growing nature of these tumors and difficulty assessing their thoracic spread, survival experiments were finished at 15 weeks (105 days).