Role of MHC Class II on tumor cells. (A) shRNA-mediated down-regulation of MHC class II expression by MB49. Three RAG.KO mice per group were challenged (day 0) with each of the tumor cell lines, MB49mock, MB49-LacZ(i), and MB49-Ab(i). On day 5, they received 8 × 105 Marilyn cells to induce the up-regulation of class II molecules and on day 13 the tumors were taken for analysis (gating on the CD45.2− and 7-AAD− population to exclude immigrating immune cells and dead cells). Histograms represent class II (Ab) staining of the tumor cells from one mouse from each group. Empty line is the isotype control. Bar graph represents the averages of the mean fluorescence of the 3 mice per group (gray area) compared with that of the isotype controls (white). (B) Marilyn cells reject tumors with knocked-down Ab: RAG.KO mice were challenged with either 105 MB49 mock cells (no symbol), or MB49-∅ (circles), MB49-LacZ(i) (squares), or MB49-Ab(i) (diamonds) cells. One day later, half of the mice in each group received 106 Marilyn cells (closed symbols). The average tumor size (10-20 mice total per group), from 3 individual experiments is shown. (C) Marilyn mice reject tumors that do not express Ab. Percentage of tumor free RAG.KO (broken line), MataHari (circles), or Marilyn (triangles) mice after challenge with 5 × 106 βTC-tet (H-2k) pancreatic tumor cells. Data were pooled from 2 experiments.