高强度聚焦超声治疗后荷瘤鼠的免疫变化及特异性抗肿瘤作用

Immune changes and specific anti-tumor immune activity in implanted H22 hepatic carcinoma after high intensity focused ultrasoundtherapy

目的:探讨高强度聚焦超声(HIFU)治疗H22移植性肝癌后的细胞免疫学变化及特异性抗肿瘤作用。方法:将C57BL/6J近交系小鼠分为HIFU治疗组、HIFU假照组和对照组,于治疗后2周流式细胞术测定各组小鼠外周血T淋巴细胞亚群的变化,采用MTT法测定脾淋巴细胞对肿瘤细胞的体外杀伤活性,ELISA法检测脾淋巴细胞与肿瘤细胞共培养24小时后上清液中IFN-γ、TNF-α的含量。结果:HIFU治疗组和对照组CD3+、CD4+T淋巴细胞,CD4+/CD8+比值明显高于假照组,CD8+T淋巴细胞低于假照组,差异有显著的统计学意义;HIFU治疗组与对照组比较,各指标均无明显差异;各组小鼠脾脏淋巴细胞对H22肿瘤细胞和S180肿瘤细胞的杀伤率,与假照组和对照组比较,HIFU组对H22肿瘤细胞的细胞毒活性明显增加,差异有显著的统计学意义,但各组对S180肿瘤细胞的杀伤率无明显的统计学差异;各组小鼠脾脏淋巴细胞与H22肿瘤细胞共培养上清液中TNF-α和INF-γ的浓度,与假照组或对照组比较,HIFU组上清液TNF-α和INF-γ浓度明显增加,差异有显著的统计学意义。结论:HIFU治疗H22移植性肝癌后,宿主细胞免疫功能明显增强,同时机体产生特异性抗肿瘤的免疫作用。

Objective：To study the immune changes and specific anti-tumor immune activity after high intensity focused ultrasound（HIFU） therapy.Methods：A total of 90 normal C57 BL/6J mice were randomly divided into three groups：HIFU-treat group,sham-HIFU group,and control group.Fourteen days after HIFU,peripheral blood was collected in each mouse to observe T lymphocytes by fluorescein activated cell sortor（FACS）.Splenic lymphocytes were also extracted to assess the cytotoxic activity against H22 and S180 tumor cells by MTT assay.Furthermore,both TNF-α and IFN-γ levels in the supernatant of culture were detected by ELISA after a 24-hour culture of spleen lymphocytes with H22 cells.Results：Compared to the values in either sham-HIFU group,there were a significant increase of CD3^＋,CD4^＋ level and CD4^＋/CD8^＋ ratios,and a significant decrease of CD8^＋ level in HIFU group and control group,but compared with the values in either HIFU group or control group,there was no significant difference of CD3^＋,CD4^＋,CD8^＋ level and CD4^＋/C D8^＋ ratios.A significant increase of splenic lymphocyte cytotoxicity on H22 tumor cells in vitro was observed in HIFU group.However,there was no significant difference splenic lymphocyte cytotoxicity to S180 tumor cells in vitro was observed in HIFU group,sham-HIFU group,and control group.Compared to the values in either sham-HIFU group or control group,there was a significant increase of both TNF-α and IFN-γ levels in HIFU group.Conclusion：The immune functions are improved in the blood after HIFU treatment for implanted H22 hepatic carcinoma in the host,and the specific anti-tumor immunity is induced by the treatment.