目的：评价多靶点抗原肽自体免疫细胞技术(multiple antigen stimulating cellular therapy, MASCTTM)治疗原发性肝细胞癌(hepatocellular carcinoma，HCC)的临床安全性。方法：回顾性分析2012年1月至2014年12月南方医院肝病中心收治的接受MASCTTM治疗且治疗前的末次治疗结束最少1个月以上、未接受其他免疫治疗的45例HCC患者的临床资料。第0天分离患者外周血单个核细胞，其中贴壁细胞诱导培养为成熟树突状细胞(muture dendritic cell，mDC)，负载14种抗原肽，小部分mDC于第8天经患者腹股沟淋巴结周围区皮下注射；未贴壁细胞第7天与大部分mDC共培养诱导为杀伤性T淋巴细胞(cytotoxic T lymphocyte,CTL)，对mDC及CTL进行质量控制检测，第26天经静脉回输患者体内，分析输注细胞后患者的不良反应和血常规、肝肾功能变化。结果： mDC中CD80+细胞比例为(98.5±5)%、CD83+细胞为(88.0±10)%、CD86+细胞为(984±3)%、HLA-DR+ 细胞为(98.8±2)%，mDC分泌高水平IL-12\[（985±312） pg/ml\]、低水平IL-10\[（53±10）pg/ml\]。CTL中CD3+CD8+细胞为（83.0±10）%、CD3+CD56+细胞为（24.0±5）%，CTL分泌高水平IFN-γ\[（1 222±650） pg/ml\]、低水平IL-10\[（7±5） pg/ml\]。MASCTTM治疗后，绝大多数患者的精神、食欲、睡眠和体力较前好转，有2名(44%)患者在输注CTL细胞1 h后出现轻中度发热，未观察到其他严重明显不良反应。患者的血常规、肝肾功能均无明显异常变化。结论： MASCTTM成功诱导培养典型mDC和CTL，行MASCTTM治疗的HCC患者不良反应较少，临床应用安全性良好

Objective:To evaluate the safety of multiple antigen stimulating cellular therapy (MASCTTM) in treating patients with hepatocellular carcinoma (HCC). Methods: The clinical datas of 45 HCC patients, who received MASCTTM from January 2012 to December 2014 in the Center of Liver Diseases of Nanfang Hospital, were retrospectively analyzed. These patients did not receive any other immunotherapy before and their latest treatment (surgery,radiotherapy or chemotherapy) was at least one month before receiving MASCTTM. Peripheral blood mononuclear cells (PBMCs) were separated on Day 0，and the adherent cells were induced into mature dendritic cells (mDCs) to load 14 types of antigen peptides, a small part of mDCs was injected intracutaneously into the area of inguinal lymph nodes on Day 8. The un-adherent cells were co-cultured with mDC on Day 7 and induced into cytotoxic T lymphocytes (CTLs); the quality of DCs and CTLs was controlled before injected intravenously into patients on Day 26. The adverse effects, blood routine examination results, and functional changes of liver and kidney were recorded after injection. Results: In mDCs, the percentage of CD80+, CD83+, CD86+ and HLA-DR+ was (98.5±5)%, (88.0±10)%, (98.4±3)%, and (98.8±2)%, respectively; mDC secreted high level of IL-12（985±312） pg/ml and low level of IL-10（53±10） pg/ml. The percentage of CD3+CD8+ and CD3+CD56+ cells in CTLs was (83.0±10)% and (24.0±5)%, respectively; CTL secreted high level of IFN-γ （1 222±650） pg/ml and low level of IL-10（7±5） pg/ml. Most of the 45 patients had alleviation in appetite, sleep and physical condition after MASCTTM treatment. There were 2 patients (4.4%) had moderate fever after infusion with CTLs, but without other severe adverse reactions. There was no significant difference in renal function or blood routine examination before and after treatment. But ATL, AST, TBIL in liver function were increased a little.Conclusion: MASCTTM induced mDC and CTL successfully; it is a feasible treatment for HCC patients with less adverse effects and good clinical safety.

国家自然科学青年基金资助项目（No.81502378); 深圳市技术创新计划创业资助项目(No.CYZZ20140421141521053)