Animal care and use were in accordance with institutional guidelines. Female FVB/N-TgN (MMTVneu)-202Mul mice (Tg-MMTVneu) (6-8 weeks old; mean weight: 18.5 g, range: 15.4-23.1 g) (Jackson Laboratory) were immunized with IGFBP-2 DNA constructs or pUMVC3 vector alone (50 μg plasmid) as a mixture in complete Freund's adjuvant/incomplete Freund's adjuvant (Sigma). Three immunizations were given two weeks apart. For tumor challenge, a syngeneic mouse mammary tumor cell line, MMC, (0.5 x 106 cells) was implanted into the mammary fat pad two weeks after the last vaccine or seven days before T-cell adoptive transfer (n=10/group) (20 (link)). Tumors were measured as previously described (16 (link)). Briefly, measurements on length, width, and depth were assessed by Vernier caliper 2-3x per week by the same technician at each time point. The measurements were multiplied by the volume constant of an ellipsoid shape, or π/6, in order to calculate tumor volumes. All tumor growth is presented as mean tumor volume (mm3 ± SEM). Data are representative of two independent experiments.
For adoptive transfer, 5x106 IGFBP-2 N-terminal- or C-terminal-specific T-cells were transferred into tumor-bearing mice by i.v. tail vein injection. The same number of splenocytes derived from naïve mice were used as a control infusion. Data are representative of two independent experiments.
For adoptive transfer, 5x106 IGFBP-2 N-terminal- or C-terminal-specific T-cells were transferred into tumor-bearing mice by i.v. tail vein injection. The same number of splenocytes derived from naïve mice were used as a control infusion. Data are representative of two independent experiments.