过继免疫超声消融后的CTL细胞在荷瘤鼠肿瘤局部的特异性表达  

Specific Expression of Active CTLs after Adoptive Immunity High Intensity Focused Ultrasound in Tumor Region

作　　者：夏纪筑[1] 丁炎[1] 曹友德[2] 伍烽[1] 

机构地区：[1]重庆医科大学生物医学工程系重庆市生物医学生物工程学重点实验室重庆市超声医学工程重点实验室一省部共建国家重点实验室培育基地,重庆市400016 [2]重庆医科大学病理科

出　　处：《中国超声医学杂志》2010年第6期481-484,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家教育部创新团队发展计划(No.IRT0454)

摘　　要：目的探讨过继免疫高强度聚焦超声(HIFU)治疗H_(22)肝癌后活化细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)在荷瘤鼠肿瘤局部的特异性表达。方法 C_(57)鼠在H_(22)肝癌移植后7d分别接受HIFU治疗和HIFU假照治疗。治疗后14 d,密度梯度离心法分离提纯2组荷瘤鼠和正常对照组脾淋巴细胞,乳酸脱氢酶(lactic dehydrogenase,LDH)法测定脾淋巴细胞对H_(22)细胞杀伤活性,同时用MHC肽五聚体荧光染色脾淋巴细胞,计数。C_(57)H_(22)荷瘤鼠随机分为HIFU组、假照组和对照组,分别接受3组染色的淋巴细胞静脉注射。输注1d后处死,取肿瘤组织作冷冻切片,激光共聚焦显微镜观察,图像软件分析。结果 HIFU组淋巴细胞对H_(22)细胞杀伤活性明显增强(P<0.05);荧光染色后,在相同数量淋巴细胞中被染的CTL细胞数,HIFU组明显比假照组和对照组多(P<0.05);3组红色荧光表达主要分布在肿瘤与正常组织交界区,肿瘤中央区未见表达;HIFU组CTL细胞在肿瘤局部的数量明显多于另两组(P<0.05)。结论 HIFU治疗H_(22)移植性肝癌后,活化的CTL细胞能到达同种荷瘤鼠肿瘤局部,发挥特异性抗肿瘤作用。Objective To investigate the specific expression of adoptive immunity active cytotoxic T lymphocyte （CTL） after high intensity focused ultrasound （HIFU） treatment of H22 hepatic carcinoma in a mouse model in tumor region. Methods C57 mice bearing H22 hepatic carcinoma received HIFU and sham HIFU exposure after tumor implantation. Fourteen days after HIFU treatment, spleen lymphocytes were extracted to assess the cytotoxic activity against H22 tumor cells by using lactic dehydrogenase （LDH） assay and to stain mouse Pro5 Recombinant MHC peptide fluorescence to count. C57 tumor mice were randomly divided into HIFU group, sham-HIFU group and control group. Each tumor mice accepted injection by the vein respectively. After 1 day, the tumor tissues were sliced into histological section, and observed with laser confocal microscope. Software Image-Pro version 5.0 was used to analyze. Results The cytotoxicity in HIFU group was stronger than in sham-HIFU group or control group （P〈0.05）. After stained the fluorescent, the amount of CTL in HIFU group was more than that in the other two groups in the same amount of lymphocytes. Special expression of fluorescent mainly distributed over the marginal region between the tumor and normal tissues, with no appearance in the central tumor. Compared to sham-HIFU group or control group, there was more CTL in tumor region in HIFU group （P〈0. 05） . Conclusions Active CTL is a specific expression in tumor local region after HIFU implanted H22 hepatic carcinoma, and plays an important role in anti-tumor.

关 键 词：高强度聚焦超声 肿瘤 过继免疫 淋巴细胞 

分 类 号：R730.5[医药卫生—肿瘤]