Selection And Treatment Of Mouse Bladder Cancer Of MUC1 Mimic Epitope Vaccine

Objective: To acquire new MUC1 mimic peptide epitopes by biospanning in phage random 12 peptide library and preliminarily investigate the functional immunological responses and effectiveness in treatment of MUC1-positive T739 mouse bladder cancer induced with the mimic peptide epitopes. To construct a recombinant plasmid expressing MUC1 antigen mimic epitope and acquire the specific purified fusion protein induced with IPTG. Based on our preliminary data, MUC1 mimic peptide epitopes will act as an important tumor-vaccine candidates for bladder cancer therapy.Methods: Monoclonal antibody BC2 which aims at MUC1 peptide epitopes was used as a ligand to screen MUC1 antigen mimic epitopes by phage display peptide library technology. Specific positive phage clones which can specifically recognize MUC1 monoclonal antibody Were screened after 3 rounds of biospanning, phage titer determination and plaque amplification in phage random 12 peptide library. Their banding capacity to BC2 antibody were tested by ELISA assay, then their DNA sequences were determined by rapid purification of sequencing templates and amino acid sequences were deducted by reduced genetic codes. Two MUC1 mimic epitopes were acquired comparing with MUC1 core sequence. Predictions of the two epitopes binding to MHC classâ… molecules and its scores evaluated the peptide's binding capacity based on the feature of motif were given from SYFPEITHI databases and competitive inhibition ELISA assay were taken to confirm the specificity of the synthetized epitope peptides. T739 mouse bladder cancer cell lines expressing human MUC1, named BST739-MUC1, were established by Lipofectamine-mediated transfection of human MUC1 full length cDNA into BST739 cells, followed by G418 selection and clonal culture. PCR, RT-PCR and immunohistochemistry were carried out to confirm the stable integration of MUC1 cDNA into the mouse genome and expression of MUC1 mRNA and proteins in the cells. The growth characteristics of...