我们前期研究发现转染AFP基因的DC疫苗可诱导抗AFP+肝癌CTL ，而对AFP-肝癌无效。因此，诱导出抗AFP-肝癌CTL成为研究者迫切解决的难题。GP73是新一代肝癌标志物，在肝癌组织中高表达，尤其在大部分AFP-表达的肝癌组织中呈阳性表达。我们推测，若将GP73和AFP联合作为靶抗原，可诱导出靶向杀伤GP73+AFP+、GP73+AFP-、GP73-AFP+肝癌细胞的CTL，对解决靶向治疗AFP-肝癌的难题有重要意义。本课题拟从患者肝癌组织中流式分选出GP73+AFP+、GP73+AFP-、GP73-AFP+、GP73-AFP-肝癌细胞，与体外诱导的患者DC融合成杂交细胞，进一步在体外诱生抗肝癌CTL，完成体外细胞毒实验，确定其对表达不同抗原的肝癌细胞的特异性杀伤作用；将CTL回输至荷人肝癌SCID鼠体内，观察其治疗作用及机制。本课题的完成，可为肝癌的免疫治聊提供新的治疗手段。

Our previous study found that the Dendrite Cell vaccine transfected AFP gene can induce CTL against AFP positive hepatoma carcinoma cell（AFP+HCC）, but not the AFP negative HCC. Therefore, how to induce anti-AFP-HCC CTL is the top difficulty concerned by many researchers. GP73 is a novel tumor marker, which is highly expressed in hepatoma carcinoma tissues, with better sensitivity and specificity than AFP in diagnosis of HCC. Especially GP73 is positive even in most AFP negative HCC patients. Accordingly, we speculated that the GP73 combined with AFP as the target antigen for immunotherapy in liver cancer, can induce CTL specifically targeting GP73+AFP+, GP73+AFP-, or GP73-AFP+HCC cells. It will be important in improving the AFP negative targeted immunotherapy. In this study, we first isolate subsets of GP73+AFP+, GP73+ AFP-, GP73-AFP+ and GP73- AFP-HCC cells by FACS and later fuse with autologous DC. The hybrid cells are co-cultured with autologous T cells to induce specific targeted CTL for cytotoxicity experiments in vitro. To study the anti-tumor effect and mechanism of specific CTLs in vivo, we first establish a mouse model by implanting the primary cultured human hepatoma cells into SCID mice which have reconstructed immune system of HCC patients. And then the induced specific CTLs aboved were infused to this animal for Anti-tumor efficacy and mechanism study in vivo.