Example 25
Prophylactic treatment of Daudi tumor Xenografts in SCID Mice with Anti CD74 HuMab Antibodies
The in vivo efficacy of anti-CD74 HuMab antibodies was determined in an intravenous (i.v.) Daudi (Burkitt's lymphoma) xenograft tumor model in SCID mice. Daudi cells were transfected by electroporation with gWIZ luciferase (Aldevron, Fargo, N. Dak., USA) and pPur vector (BD Biosciences, Alphen a/d Rijn, The Netherlands) in a 4:1 ratio. After 48 h, puromycin was added for selection of a stably transfected clone (Daudi-luc). Daudi luc #1E3 cells were cultured in RPMI supplemented with 10% cosmic calf serum (cat. no. SH30087.04, Hyclone), 1% penicillin/streptomycin (cat. no. DE17 603, Cambrex, Germany), 1% sodium pyruvate and 1 μg/mL puromycin (cat. no. P 8833, Sigma, Zwijndrecht, The Netherlands). 2.5×106 Daudi luc tumor cells in 100 μL PBS were injected i.v. in the tail vein of female SCID mice (7 mice per group). Mice were treated at the day of tumor inoculation with 100 μg (˜5 mg/kg) HuMab-CD74-005, -006 or -011 or control antibody (IgG1b12), in 200 μL PBS, intraperitoneally (i.p.). Mice were imaged directly after tumor inoculation, followed by imaging at weekly intervals starting on day 14. For imaging, mice were anesthetized using isoflurane, followed by i.p. administration of 2.5 mg D luciferin (acid form, cat. no. BT11 1000; Biothema, Haninge, Sweden) in 200 μL 10 mg/mL TRIS (cat. no. T60666-1 kg, Sigma). Bioluminescence imaging (BLI), from the back side (dorsal view), started 10 min after administration of D luciferin, 5 min exposure time, on a Biospace Imager. Black and white images were made for anatomical reference.
