Figure 3. Immunization of WT but not MUC1.Tg mice with MUC1p results in up-regulation of pancreatic enzymes in splenic DC.

(A) WT and MUC1.Tg mice were injected i.v with unloaded BMDC (ctrl) or BMDC loaded with MUC1 peptide (MUC1p). 24h later spleens were harvested, pooled according to group, and RNA extracted for qRT-PCR. Arbitrary Units were normalized to WT mice given the ctrl vaccine. Bars represent mean ± SEM. Data are representative of two independent experiments. (B) Splenic DC from unvaccinated mice were isolated with CD11c⁺ beads (n=3), total splenic T cells were isolated using negative selection via MACS depletion of CD3⁻ cells, and BMDM (MΦ) were cultured for 8 days in the presence of L-cell supernatant as a source of M-CSF. RNA was isolated from all populations for qRT-PCR analysis. Units were normalized to expression levels in CD11c⁺ cells.Bars represent mean ± SEM. Data representative of two independent experiments. (C) WT and MUC1.Tg mice were immunized as in (A). At 24h, splenic DC were isolated using CD11c⁺ beads for analysis by qRT-PCR or Western blotting for trypsin and CPB1 (D). Bars represent mean ± SEM after normalization to control vaccination. Data are representative of two (C) and three (D) independent experiments. (E) Mice were immunized i.v. with PBS (ctrl), Poly-ICLC (adj), or soluble MUC1p admixed with Poly-ICLC (MUC1p + Adj). 24h later spleens were harvested for qRT-PCR analysis. Bars represent mean ± SEM normalized to PBS control and are representative of four independent experiments.