The Study Of Anti-tumor Ability Of DC Pulsed By TRAP

In this study, We developed a protocol to acquire DCs from human spleen.The tumor antigen peptide of human hepatocarcinoma cell line HHCC was used to pulse DCs .Then navie T cells of the same person was actived by the DC vaccine. We inoculated them into nude mice to observe their inhibition to HHCC transplanted tumor on nude mice. Human spleen cells were acquired by the general method. Lymphocytes were isolated on lymphocyte separating solution. The low density cells were collected and depleted of unadherent cells after 4h culture. The M?s which closely adhered the bottom of culture bottles were discarded . The DCs-enriched population was acquired by maintained in RPMI 1640 supplemented with recombinant human GM-CSF (500U/ml) >recombinant human TNF- a (500U/ml) x mouse anti human CD40 McAb (Ing/ml) for 7-9 days. DCs were observed with SEM. They were identified by their typical morphological feature of long protrusions. The percentage of DC contained and their surface molecules were counted by Flow Cytometry (FCM) EPICS-PROFILE and immunohistochemistry .After the actived T and peptide pulsed DC were inoculated into nude mice, We kept on measuring the volume of nude mice's implanted tumor. Their survival term were also wrote down. With this protocol, the percentage of DCs arrived 98 %. The number of double positive cells with both CD40 and HLA-DR molecules rised with their cultured day. The DCs were potent stimulators in MLR. The experiment5of mice showed that the induced CTL inhibited the growth of implanted tumor in nude mice and protected the nude mice against the further challenged of HHCC tumor cells. As a new concept anti-tumor vaccine, DC pulsed by tumor antigen peptide may play certain role in treatment and prevention of tumor.