Figure 3.

Measurement of proliferative responses by CFSE dilution after immunization/challenge. The differences in cell proliferation (%) were calculated by subtracting values obtained in assays using spleen cells from FSWT-immunized mice from those obtained in assays using spleen cells from FSVL-immunized animals (A, B and C). (A) Mice were immunized with FSVL and FSWT, 40 day later the spleen cells were harvested and stimulated for 72 hours with the panel of 87 epitope variant-bearing phage clones, as well as non-related FB22 and SWT-expressing FSWT phage. (B) 40 days after priming, mice were inoculated with 4T1 tumor cells and, spleen cells collected on day 15 were stimulated as described above. (C) Mice were challenged with tumor cells and immunized with FSVL and FSWT on day 5, then on day 15 spleen cells were processed for cell proliferation. (D) Differences in cell proliferation (%) between non-immunized tumor-challenged and naïve mice. The spleen cells from untreated (tumor free) or 4T1-inoculated mice were harvested on day 15 post-tumor challenge and used in cell proliferation. The vertical red lines indicate arbitrary defined 3 % threshold for differences in cell proliferation. Data from one representative experiment out of 2 are shown.