The recombinant fowlpox virus containing the gene for murine GM CSF has also been described previously. Recombinant fowlpox infections and recombinant vaccinia containing murine B7 1, ICAM 1, and LFA 3 genes in conjunction with human CEA have now been described previously. The recombinant HDAC1 inhibitor fowlpox virus containing the gene for murine GM-CSF has also been described previously. Proteins H 2Db restricted influenza virus A/NT/60/68 peptide, influenza virus A/PR/8/34 and H 2Db restricted CEA peptide were produced by CPC Scientific. In vitro assay Primary splenocytes were dispersed into single cell suspensions, the red blood cells were eliminated by lysis, and the remaining cells seeded into 6 well plates at 6 105 cells/ml in complete RPMI media. Splenocytes from mice were stimulated with 10 ug/ml of soluble anti CD3e and splenocytes from mice were stimulated with 10 4 ug /ml of NP68 peptide then used in the appropriate experiments. For western blot analysis and kinase assay, cells were collected at the indicated time points and the CD8 T cells were chosen using magnetic cell sorting. Ex vivo assays Primary splenocytes from both vaccinated or naive C57BL/6 Digestion rats were dispersed into single cell suspensions followed closely by removal of red blood cells, and 5 106/ml cells were cultured in 1. 6 ml complete RPMI containing 1 ug/ml of cognate peptide with or without 10specific pathogen-free conditions and in accordance with the Association for Assessment and Accreditation of Laboratory Animal Care tips. All experimental studies were completed beneath the agreement of the Intramural Animal Care and Use Committee. Cell Lines Murine colon carcinoma MC38 cells expressing human CEA were created by retroviral transduction with CEA cDNA. MC32a cells were cultured in MEM medium supplemented with 1 non-essential amino acids, 1 mmol/L sodium pyruvate, 2 mmol/L L glutamine, 10 mmol/L HEPES, 300 ug/mL G418 sulfate, and ten percent heatinactivated fetal bovine serum. Their components and ARN-509 structure all media were purchased from Mediatech, unless otherwise indicated. Inhibitors For in vitro research, saracatinib or dasatinib were dissolved in dimethyl sulfoxide, and diluted in culture media into a respective final concentration. The optimum concentration of DMSO was 0. 10 percent. As a 1 mg/ml alternative and dasatinib was formulated as a 0 for in vivo study, saracatinib was formulated. 25 mg/ml solution in water with one of the tween 80. These solutions were given orally through the use of plastic feeding tube. Poxvirus constructs Recombinant vaccinia and recombinant fowlpox worms containing murine B7 1, ICAM 1, and LFA 3 genes in combination with nucleoprotein of influenza virus A/PR/8/34 have now been described previously. Recombinant vaccinia and recombinant fowlpox infections containing murine B7 1, ICAM 1, and LFA 3 genes in combination with human CEA have now been described previously.