目的：评价活化自体淋巴细胞过继性免疫治疗（adoptive immunotherapy，AIT）是否有助于改善原发性肝细胞癌的临床疗效。方法：选取2016年8月至2018年12月在中国人民解放军总医院第五医学中心确诊的64例原发性肝细胞癌患者，通过分层随机法分为免疫治疗组（n=29）和对照组（n=35）。免疫治疗组患者取60 ml外周血分离制备单个核细胞并在含OKT-3和IL-2的培养基中活化培养，回输前进行质控检测。免疫治疗组中的Ⅰ~Ⅲ期患者（n=14）于一线治疗后接受自体淋巴细胞输注（3个月内输注6次），Ⅳ期患者（n=15）仅接受自体淋巴细胞输注；对照组患者接受肝细胞癌相关的其他治疗。疗效评估的主要终点是2 年无复发生存（relapse-free survival，RFS）率，次要终点为无进展生存期（progression-free survival，PFS）和总生存期（overall survival，OS）。结果：入组患者中位随访时间为2.8年（0.2~4.2年）。免疫治疗组29名患者共接受了167次（计划174次，完成率96%）预定淋巴细胞输注（平均每人次回输9.30×109个细胞，其中CD3+HLA-DR细胞约占63%），治疗期间未观察到3级或4级不良反应发生。与对照组相比，免疫治疗组患者2年RFS率显著升高（62.1% vs 22.9%，OR=0.181，95%CI：0.06~0.54，P=0.002），中位PFS（28 vs 8个月，P=0.004）和中位OS（38 vs 34个月，P=0.915）均显著延长。在Ⅰ~Ⅲ期患者中，免疫治疗组（n=14）2年RFS率较对照组（n=18）显著升高（92.9% vs 33.3%，OR=0.38，95%CI：0.004~0.368，P=0.005），中位PFS明显延长（38 vs 14.5个月，P=0.005），而两组OS间无显著差异；Ⅳ期患者两组间PFS（P=0.077）及OS（P=0.994）均未见显著差异。结论：活化自体淋巴细胞AIT为安全可行的肝细胞癌辅助性治疗方法，可提高Ⅰ~Ⅲ期肝细胞癌一线治疗后RFS率、延长患者RFS时间，而对进展期肝细胞癌患者的PFS和OS无明显影响。

Objective: To evaluate whether adoptive immunotherapy (AIT) with activated autologous lymphocytes helps to improve the clinical efficacy of primary hepatocellular carcinoma. Methods: Sixty-four patients with primary hepatocellular carcinoma diagnosed at the Fifth Medical Center of Chinese PLA General Hospital from August 2016 to December 2018 were enrolled and divided into immunotherapy group (n=29) and control group (n=35) by the stratified randomized sampling method. 60 ml of peripheral blood was drawn from each patient in the immunotherapy group to prepare mononuclear cells, which were then activated and cultured in a medium containing OKT-3 and IL-2. A quality control test was done before blood re-transfusion. In the immunotherapy group,patients in stage Ⅰ-Ⅲ (n=14) received autologous lymphocyte infusion (6 infusions within 3 months) after first-line treatment, while patients in stage Ⅳ (n=15) only received autologous lymphocyte infusion therapy. Patients in the control group received other hepatocellular carcinoma-related treatments. The primary endpoint of the efficacy evaluation was the 2-year relapse-free survival (RFS), and the secondary endpoints were progression-free survival (PFS) and overall survival (OS). Results: The median follow-up time of the enrolled patients was 2.8 years (0.2-4.2 years). 29 patients in the immunotherapy group received a total of 167 scheduled lymphocyte infusions (with an average of 9.30×109 cells per patient, of which CD3+HLA-DR cells accounted for about 63%; 174 infusions were planned, with a completion rate of 96%). During the treatment period, no grade 3-4 adverse reactions were observed.Compared with the control group, patients in the immunotherapy group had a significantly increased 2-year RFS rate (62.1% vs 22.9%,OR=0.181, 95%CI: 0.06-0.54, P=0.002), and the median PFS (28 vs 8 months, P=0.004) and median OS (38 vs 34 months, P=0.915) were significantly prolonged. Among the patients at stage Ⅰ-Ⅲ, the PFS rate in the immunotherapy group was significantly higher than that in the control group (92.9% vs 33.3%, OR=0.38, 95%CI: 0.004-0.368, P=0.005), and the median PFS was significantly prolonged (38 vs 14.5 months, P=0.005), but there was no significant difference in OS between the two groups. In stage Ⅳ patients, there was no statistical difference in PFS (P=0.077) and OS (P=0.994) between the two groups. Conclusion: AIT with activated autologous lymphocytes is a safe and feasible adjuvant therapy for hepatocellular carcinoma, which can improve RFS rate after first-line treatment of stage Ⅰ-Ⅲ hepatocellular carcinoma and prolong the recurrence-free survival time of patients. It has no significant effect on PFS and OS in patients with advanced hepatocellular carcinoma.

军队后勤科研重大项目资助（No. AWS17J010）