微波热消融后活化的淋巴细胞在同源肿瘤效应的实验研究

Effects of microwave ablation-activated lymphocytes after adoptive immunotherapy in homogous tumors

目的探讨微波热消融治疗H22小鼠肝癌细胞移植荷瘤小鼠后活化的淋巴细胞,过继免疫给另一组同源荷瘤小鼠,体外观察其抗肿瘤效应。方法成年健康C57BL/6J近交系荷瘤小鼠90只,雌雄不限,7~8周龄,体质量20~25 g。取60只,随机分治疗组（n=20）、假照组（n=20）和正常对照组（n=20）;在H22小鼠肝癌细胞移植术后7 d,治疗组接受微波热消融治疗,假照组不启用微波热消融,正常对照组不做任何处理;治疗后10 d,分离2组荷瘤小鼠和正常对照组荷瘤小鼠脾淋巴细胞。另选30只C57BL/6J荷瘤小鼠,随机分为过继免疫治疗组（n=10）、过继免疫假照组（n=10）和过继免疫对照组（n=10）,前2组用H22小鼠肝癌细胞移植造模。7 d后,3组分别经鼠尾静脉注射之前提取的淋巴细胞。输注7 d后将肿瘤病灶制成单细胞悬液,测定肿瘤局部淋巴细胞的功能变化。结果与假照组和正常对照组比较,治疗组对H22小鼠肝癌细胞杀伤活性明显增加,差异有显著统计学意义（P〈0.01）。淋巴细胞过继免疫治疗后7 d,与过继免疫假照组和过继免疫对照组比较,过继免疫治疗组分泌的淋巴细胞数量明显增多,差异有显著统计学意义（P〈0.01）。结论微波热消融治疗H22小鼠肝癌细胞移植后,活化的淋巴细胞在同种来源肿瘤发挥特异性的抗肿瘤效应。

Objective To investigate the functional changes of thermal ablation-activated treatment for lymphocytes transplanted adoptive immunotherapy mice from H= cognate tumor-bearing mice, and observe the anti-tumor effects in vitro. Methods Ninety adult healthy C57BL/6J inbred tumor- bearing mice were sacrificed, male or female, aged 7 - 8 weeks old with body weight of 20 - 25 g. Sixty of mice were randomly divided into treatment group（n = 20）, sham irradiation group（n = 20） and normal control group（n = 20）; After H= hepatoma transplantation for 7 days, treatment group received microwave thermal ablation, sham irradiation group was given un-enabled microwave thermal ablation and normal control group without any treatment. After treatment for 10 days, spleen lymphocytes of 2 groups of tumor-bearing mice and normal control group mice were separated. Thirty C57BU6J inbred tumor-bearing mice were randomly divided into adoptive immune therapy group（n = 10）, adoptive immunity sham irradiation group （n = 10） and adoptive immune control group （n = 10）, respectively, the first 2 groups were transplanted with H22 hepatoma cells, which were injected on caudal vein with prior extracted lymphocytes ceils. The tumor lesion was made into unicellular suspension and functional changes of tumor cells were measured at 7 days after the treatment. Results Compared with sham irradiation group and normal control group, H22 hepotama cell killing activity in microwave thermal ablation group was significant increased, the difference was statistically significant（P 〈 0.01）; Compared with adoptive immunity sham irradiation group and adoptive immune control group, lymphocytes in adoptive immune therapy group increased significantly after adoptive immunotherapy for 7 days, and the difference was statistically significant （P 〈 0.01）. Conclusion It is demonstrated that after microwave thermal ablation treatment of H22 transplanted liver, the activated lymphocytes play specific anti-tumor effects in isotype source tumor.