肝组织内序贯注射灭活异体淋巴细胞和自体淋巴细胞治疗小鼠肝癌移植瘤的研究  

Treatment of transplanted hepatocarcinoma in mice with sequential injection of inactivated heterogeneic lymphocyte and autogeneic lymphocyte

作　　者：吴林岚[1] 徐兵 黄素钦 魏建威[1] 杨晓梅 赵芝萍 陈怡 蒋晓织 WU Lin fan;XUBing;HUANG Su qin;WEI Jian wei;YANG Xiao mei;ZHAO Zhi ping;CHEN Yi;J IANG Xiao zhi(Clinical Laboratory of Second people＇s Hospital of Fujian Province,Fuzhou 350003,China)

机构地区：[1]福建省第二人民医院检验科,福州350003 [2]福建医科大学孟超肝胆医院肝病研究所 [3]福州市菁华崇辉医院肝内科

出　　处：《肝脏》2018年第5期387-390,共4页Chinese Hepatology

基　　金：福建省医学创新课题研究项目(2017-CXB-12)

摘　　要：目的探讨用异体淋巴细胞(heterogeneic lymphocyte,HL)和自体淋巴细胞(autogeneic lymphocyte,AL)序贯注射的抗肿瘤方法。方法取供鼠C3H小鼠脾淋巴细胞,用丝裂霉素灭活制备灭活异体淋巴细胞(inactivated HL,IHL)。在受鼠CB6F1小鼠肝组织内接种hepa1-6瘤细胞。用冻融法从小鼠血浆提取冷沉淀,制成纤维蛋白胶(fibrin glue,FG);用FG与IHL或AL组合成FG-IHL或FG-AL。前阶段:用FG-IHL注射到荷瘤受鼠肝内肿瘤接种部位治疗(实验组);用FG-PBS同法注射受鼠肝内肿瘤接种部位作为对照组。而后检测两组受鼠的脾淋巴细胞杀瘤细胞率和脾淋巴细胞、CD8^+T淋巴细胞、NK细胞数量等免疫学指标。后阶段:从实验组、对照组中随机选出部分受鼠,取淋巴细胞制成FGAL,将各组FG-AL分别注射到本组其余受鼠的肝内肿瘤接种部位;治疗后剥出受鼠肝内肿瘤,比较两组的瘤体积和抑瘤率。结果前阶段治疗后,实验组受鼠AL的杀瘤细胞率为(25.7±4.81)%,明显高于对照组AL的相应值(E∶T=60∶1,P<0.01);实验组受鼠脾淋巴细胞、CD8^+T细胞和NK细胞数量均明显高于对照组的相应值(均P<0.05)。经前、后两阶段治疗后,实验组受鼠的瘤平均体积为(1.15±0.25)cm^3,明显小于对照组的瘤平均体积为(1.91±0.37)cm^3(P<0.01);实验组的抑瘤率为39.8%(以对照组作基准)。结论肝组织内肿瘤局部注射IHL,再注射AL的序贯疗法,可明显抑制小鼠移植肝肿瘤生长,有望成为一种抗肿瘤生物疗法。Objective To investigate the anti-tumor effect of sequential injection of heterogeneic lymphocyte（HL）and autogeneic lymphocyte（AL）.Methods Inactivated heterogeneic lymphocyte（IHL）was isolated from spleen of C57 BL/6 mice and inactivated with mitomycin.Hepa1-6 cells were inoculated into liver tissue of CB6 F1 mice.Fibrin glue（FG）was prepared with cryoprecipitate collected from mouse plasma using freeze-thaw method.The experimental treatment consisted of two stages.At first stage（13 days）,tumor-bearing mice were injected with FG-IHL in liver as experiment group,and injected with FG-phosphate buffer saline（FG-PBS）in liver as control group.Immunological indices such as tumor cell killing rate of the spleen lymphocytes and number of lymphocytes,CD8＋T and NK in the two groups were detected,respectively.At the later stage（8 days）,FG-AL was prepared from randomly selected tumor-bearing mice in experiment and control group,which was injected into the liver of rest tumor-bearing mice in the same group,respectively.After 8-day treatment,tumor size and inhibition rate were compared between the two groups.Results The tumor cell killing rate of AL in experiment group was significantly higher than that in control group（25.7±4.81% vs 5.9±1.64,P〈0.01）.Numbers of spleen lymphocyte,CD8＋T and NK cell in experiment group were significantly higher than those in control group,respectively（all P〈0.05）.After 2-stage treatment,mean volume of tumor in experiment group was significantly smaller than that in control group（1.15±0.25 cm3 vs.1.91±0.37 cm3,P〈0.01）.Tumor inhibiting rate in experiment group was 39.8%.Conclusion The treatment of sequential injections with FG-IHL followed by the FG-AL in situ could suppress the growth of transplanted tumor in mice,which might become a promising biotherapy to treat cancers.

关 键 词：淋巴细胞 注射 肝癌 纤维蛋白胶 

分 类 号：R735.7[医药卫生—肿瘤]