MUC1VNTR核酸疫苗的优化构建及其抗胰腺癌的实验研究.pdfVIP

9220 2013 10 7 20 Chin J Clinicians Electronic Edition ,October 15,2013,Vol.7,No.20 MUC1-VNTRn MUC1-VNTRn pVAX1-MUC1-VNTRn pVAX1-MUC1-VNTRn T ELISPOT IFN-γT CTL CytoTox? MUC1-VNTRn CTL Capan-2 pVAX1-MUC1-VNTRn pVAX1-MUC1-VNTRn pVAX1-MUC1-VNTRn CD80 CD86HLA-DR IL-12 P 0.01 MUC1-VNTR MUC1-VNTR T IFN-γ T 103.08.5 6 9 94.37.7P 0.001MUC1-VNTR MUC1-VNTR CTL Capan-2 6 9 40.123.16%37.313.95%MUC1-VNTR VNTR VNTR VNTR MUC1-cDNA 1 1 3 4 P 0.001 pVAX1-MUC1-VNTR6 panc02-MUC1 pVAX1-MUC1-VNTR VNTR VNTR P 0.01 pVAX1-MUC1-VNTRn 1 3 9 pVAX1-MUC1-VNTR6 DNA CA-15-3 Optimized construction of MUC1-VNTRn DNA vaccine and the anti-pancreatic cancer research ZHOU Quan-bo, GONG Yuan-feng, ZHOU Yu, LIN Qing, LI Zhi-hua, CHEN Ru-fu, ZENG Bing, ZHENG Li-ping. Department of Hepatobiliary Surgery , The Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China Corresponding author: CHEN Ru-fu, Email: chenrf10@? Abstract Objective MUC1-VNTRn served as a new target for pancreatic cancer immunotherapy in the present study. We intended to screen out the most powerful pVAX1-MUC1-VNTRn DNA vaccine which showed strongest immunogenicity in vitro and vivo. Methods pVAX1-MUC1-VNTRn were constructed and transfected into MUC1 negative cells. The target protein of MUC1-VNTRn could be detected by Western Blot. Dendritic cell preparation and research of biological characteristics. pVAX1-MUC1-VNTRn were transfected into dendritic cell. The stimulated mature dendritic cells were co-cultured with autologous T cells, and then we used the ELISPOT to detect the IFN-γ secreting CTL. We detected the cytotoxic effect of the antigen-specified CTL to Capan-2 cells in vitro. Finally, we used the the protective and therapeutic immune response in mice to confirm the immunogenicity of DNA vaccine. Results pVAX1-MUC1-VNTRn were successfully constructed, and the target protein can be expressed in eukaryotic cells. The immature de