树突细胞联合细胞因子诱导的杀伤细胞对原发性肝脏淋巴瘤生长和术后复发转移的抑制作用  

Inhibitory effect of dendritic cells plus cytokine induced killer cells on the growth and postoperative recurrence and metastasis of primary liver lymphoma

作　　者：脱帅 张宁苏[2] 张宁 杨波[4] 

机构地区：[1]解放军202医院肝胆外科,沈阳110812 [2]辽宁中医药大学附属医院血液肿瘤科,沈阳110032 [3]解放军202医院肿瘤超微病理研究室,沈阳110812 [4]解放军总医院南楼血液科,北京100853

出　　处：《解放军医学杂志》2010年第12期1449-1454,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家重点科技攻关计划项目(96-A23-06-03);辽宁省自然科学基金(20042153)

摘　　要：目的探讨树突细胞(DC)与细胞因子诱导的杀伤细胞(CIK)共培养后产生的DC-CIK细胞对原发性肝恶性淋巴瘤生长和术后复发转移的抑制作用。方法制备健康人和原发性肝淋巴瘤患者来源的DC和CIK细胞,共培养后获得DC-CIK细胞。取上述原发性肝淋巴瘤患者的恶性淋巴瘤组织,采用外科原位移植技术建立人原发性肝恶性淋巴瘤裸鼠原位移植模型HLBL-0102。另外,在此模型基础上,通过根治性切除术,建立术后肝淋巴瘤肝内复发转移裸鼠模型HLBL-0701。将以上2种模型裸鼠按照以下干预措施分为7组,分别给予CHOP方案化疗、健康人CIK细胞输注、健康人DC-CIK细胞输注、肝淋巴瘤自体CIK细胞输注、肝淋巴瘤自体DC-CIK细胞输注、肝淋巴瘤自体DC-CIK输注联合CHOP方案化疗,以及生理盐水对照。CHOP方案和生理盐水为0.3ml/(kg·d),CIK或DC-CIK细胞均以6×107/(kg·d)[约0.3ml(kg·d)]输注。连续干预21d,末次用药后72h,经心脏采血,测定乳酸脱氢酶(LDH)浓度。取死亡裸鼠,测量肿瘤体积,计算抑瘤率和肝内复发转移率。分析DC-CIK细胞治疗与化疗的协同作用。结果在HLBL-0102模型中进行的实验表明,与对照比较,化疗,健康人CIK、DC-CIK细胞,肝淋巴瘤自体CIK、DC-CIK细胞,以及肝淋巴瘤自体DC-CIK细胞+化疗干预后的抑瘤率分别为52.1%、40.2%、54.3%、41.7%、60.6%和84.3%,而在HLBL-0701模型中,上述干预措施对肝内肿瘤复发转移的抑制率分别为75.0%、75.0%、50.0%、62.5%、37.5%和25.0%;干预后裸鼠平均生存时间分别为70.32±6.54d、68.25±5.37d、77.00±6.37d、75.25±5.80d、79.50±8.07d、84.75±8.26d,显著长于对照组(35.00±4.17d,P<0.01)。在HLBL-0102模型中,瘤体积与LDH浓度呈显著正相关(r=0.9898,P<0.05),协同分析证实肝淋巴瘤自体DC-CIK细胞与化疗具有相加作用(q=1.038)。结论共培养DC-CIK细胞输注能抑制裸鼠原发性肝淋巴瘤生长和肝淋巴瘤根治切除术后的复发转移Objective To explore the inhibitory effect of DC-CIKC cells yielded by co-cultivation of dendritic cells（DC）and homologous cytokine induced killer cells（CIKC）on the growth and postoperative recurrence and metastasis of primary liver lymphoma. Methods The peripheral blood mononuclear cells（PBMCs）from healthy donors and a patient with primary liver lymphoma were isolated and separately cultured ex vivo to prepare DC and CIKC cells.The prepared DC and CIKC cells were then co-cultured for 14days in the proportion of 1：5to yield DC-CIKC cells.The resected liver tumor specimens from the patient mentioned above were implanted into the liver of nude mice by surgical orthotopic implantation to establish the orthotopic nude mouse model of human primary liver lymphoma, named HLBL-0102.On the basis of HLBL-0102model,recurrent and metastatic nude mouse model of primary liver lymphoma,designated HLBL-0701,was reproduced by radical resection of primary tumor of HLBL-0102model.The HLBL-0102and HLBL-0701models were randomly and averagely assigned into seven groups（8each）：CHOP chemotherapy regimen group,healthy donor CIKC group,healthy donor DC-CIKC group,patient CIKCC group,patient DC-CIKC group,patient DC-CIKC plus CHOP chemotherapy group,and control group.Animals in control group the animals were given physiological saline of 0.3ml/（kg·d）,in chemotherapy group,the animals were given CHOP regimen（cyclophosphamide 100mg,adriamycin 40mg,oncovin 2mg,dexamethasone 5mg）of 0.3ml/（kg·d）,and in animals of CIKC and DC-CIKC groups were administered with effector cells in the dosage of 6 107/0.3（kg·d）.All the treatment lasted for 21 days.Cancer-bearing nude mice were sacrificed 72hours after last treatment.Blood samples were collected from heart for lactate dehydrogenase（LDH）detection.The tumors were removed to measure the weight,and the longest and shortest diameters of tumors.The volume of tumors and inhibitory rate of tumor weight were calculated,respectively.Both light and electron microscopy

关 键 词：树突细胞 杀伤细胞 肝肿瘤 实验性 免疫疗法 

分 类 号：R735.7[医药卫生—肿瘤]