食管癌疫苗的研制及临床应用效果观察

Preparation and clinical application of esophagus cancer vaccine

目的探讨食管癌疫苗的抗癌机制及临床应用效果。方法培养食管癌细胞Ecal09，提取其抗原成分和超抗原金黄色葡葡球菌肠毒素C型构建成肿瘤疫苗；分离人外周血单个核细胞（PBMC），与肿瘤疫苗联合作用，进行体外培养增殖，成为效应细胞；流式细胞仪测定效应细胞表型；细胞毒实验检测其杀伤活性。选取早期食管癌术后患者106例，分为观察组53例，应用食管癌疫苗治疗，对照组53例实施常规治疗。观察对比临床应用效果，对两组患者进行3年随访观察其生存率。结果经肿瘤疫苗刺激的淋巴细胞组增殖活性最强，于72h达到高峰（A值为0．22）；并且上调细胞毒性T细胞（CTLs），CTLs对靶细胞杀伤活性外周血单个核细胞＋超抗原金黄色葡萄球菌肠毒素C型＋肿瘤可溶性抗原组（97．36-2．11）％显著高于单个核细胞组（79．27-5．57）％，差异有统计学意义（P〈0．01）。3年随访发现观察组生存率[男48．28％（14／29）、女45．83％（11／24）]高于对照组[男21．42％（6／28）、女24．00％（6／25）]，差异有统计学意义（χ^2=5．06、x。6．28，P均〈0．05）。结论食管癌疫苗能诱导效应细胞明显增殖、活化、并产生高效、特异的抗肿瘤效果。临床观察发现其能有效防止食管癌术后患者的复发和肿瘤转移，提高生存率。

Objective To explore the anticancer mechanism of esophageal cancer vaccines and clinical effect. Methods Esophagus cancer cells （Eca109） were cultured and the soluble antigen were extracted from the cells. Esophagus cancer vaccine was constructed with the antigen and superantigen SEC. Lymphocytes were isolated from peripheral blood and then stimulated with the vaccine in vitro. Phenotypes of the cells were checked by FCM and killing activity was tested with cytotoxic assays. One hundred and six early esophageal cancer patients were selected after surgery, who were divided into the observation group of 53 cases with esophageal cancer vaccine therapy and 53 cases in the control group with conventional treatment. Then clinical effect was observated and 3 year follow-up survival rate was observed of the of two groups of patients. Results Proliferation of vaccine stimulated lymphocyte group was the strongest,and high peaked at 72 h（A =0. 22）,and raise CD8- T cell populations of CTLs. The killing activity of lymphocyte group stimulated by the vaccine against target cells was significantly higher than that of lymphocyte group （ （97. 36＋2. 11）% ） vs. （79. 27-5. 57）%,F = 38. 6：2 ,P〈0.01 ）. Three years follow up shows that the survival rates of experiment group were 48. 27% （male） and 45. 83% （female） respectively, and control group were 21.43% （male） and 24. 00% （ female ）, and the difference was significant （χ^2 = 5.06, 6. 28, P 〈 0.05 ）. Conclusion The tumor vaccine constructed with esophagus cancer antigen and superantigen SEC can induce PBMC to activate and proliferate into CD8- CTL with specific cytotoxicity against the cells which the antigen comes from. The vaccine may raise the survival rate of the patients with Esophagus cancer.