卵巢癌特异性免疫细胞治疗的体内外实验研究  

Specific Immune Cell Therapy Against Ovarian Cancer In Vivo and In Vitro

作　　者：昌晓红[1] 程洪艳[1] 成夜霞[1] 叶雪[1] 郭慧方[1] 付天云[1] 张丽[1] 张果[1] 崔恒[1] 

机构地区：[1]北京大学人民医院妇科肿瘤中心,北京100044

出　　处：《癌症》2008年第12期1244-1250,共7页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.30571940)~~

摘　　要：背景与目的:卵巢癌抗独特型微抗体(6B11mini)是部分人源化的抗独特型卵巢癌疫苗,体内外实验研究证明,它可以诱导出特异的抗卵巢癌体液免疫和细胞免疫。本研究评价将6B11mini作为抗原,采用免疫细胞的方法处理卵巢癌时的安全性和有效性。方法:用纯化的6B11mini负载树突细胞(dendritic cell,DC),与外周血单个核细胞(peripheral blood mononucleocyte,PBMNC)混合培养,在多种细胞因子共同作用下获得效应细胞6B11-OCIK。通过软琼脂克隆形成实验及接种裸鼠皮下瘤实验,观察6B11-OCIK在体内和体外的成瘤能力;将6B11-OCIK静脉注射BALB/c小鼠,观察急性毒性反应;体外Cr释放实验检测6B11-OCIK对靶细胞的杀伤作用。51用人卵巢癌细胞株SKOV3构建严重联合免疫缺陷(severe combined immune deficieney,SCID)小鼠卵巢癌移植瘤模型,注射6B11-OCIK,并以CIK、PBMNC细胞以及生理盐水作为对照组,分别观察各组肿瘤生长情况。结果:软琼脂培养14d,卵巢癌SKOV3细胞克隆形成良好,克隆形成率50%;皮下接种裸鼠后14d,阳性对照的宫颈癌HeLa细胞组全部成瘤,6B11-OCIK、CIK、WI-38组和新鲜PBMNC组持续观察13周没有成瘤。6B11-OCIK静脉注射BALB/c小鼠,30min内各剂量实验组和生理盐水对照组动物都无任何明显的不良反应;输注后第13天处死小鼠,解剖小鼠观察其各主要脏器无明显异常。在体外杀伤实验中6B11-OCIK对抗原阳性的肿瘤细胞具有特异性杀伤作用,并且与MHC限制性相关;在荷瘤SCID小鼠中6B11-OCIK治疗组肿瘤重量与生理盐水对照组相比差异有统计学意义(P=0.023),与CIK组、新鲜单采组相比差异无统计学意义(P=0.540,P=0.285)。结论:6B11-OCIK在动物体内应用时符合安全性指标,并对卵巢癌的生长有一定的抑制作用。BACKGROUND＆OBJECTIVE： 6B11minibody （6B11mini）, an anti-idiotypic vaccine against human ovarian cancer, has been proven to induce specific humoral and cellular immunity against ovarian cancer in vivo and in vitro. This study was to investigate the safety and efficacy of using 6B11mini as an antigen to treat ovarian cancer. METHODS. After being loaded with purified 6B11mini, dendritic ceils （DCs） were co-cuLtured with peripheral blood mononucleocytes （PBMNC） and stimulated by various cytokines, including CD3 monoclonal antibody,interleukin-2, interferon-γ, to obtain 6Bllmini-ovarian-cytokine-induced-kilier ceils （6B11-OCIK）. Tumorforming ability was determined using soft agar colony-forming assay in vitro and nude mice xenografts in vivo. The acute toxicity of 6B11-OCIK at different doses was observed in BALB/c mice. Cytotoxicity of 6B11-OCIK to different target cells was detected using ^60Cr release test in vitro. The ovarian tumor model was established using severe combined immune deficiency （SCID） mice transplanted with human ovarian cancer cell line SKOV3. The tumor growth was detected after injection of 6B11-OCIK into SCID mice. Injection of CIK, PBMNC and physiological saline were used as controls. RESULTS： After a cultual period of 14 days in soft agar, SKOV3 cell clones were well formed with a ratio of 50%; while 6B11-OCIK, CIK and PBMNC did not form any clones. All nude mice injected with human cervical carcinoma cell line HeLa （positive control） grew tumors after 14 days, and mice injected with 6B11-OCIK, CIK, PBMNC and normal human fetal lung fibroblast Wl-38 cells did not form tumors after 13 weeks. BALB/c mice did not show any abnormal response half an hour after the administration of 6B11-OCIK cells at different doses. Mice were sacrificed 13 days after treatments, but no distinct abnormity of the main organs were found. 6B11- OCIK exerted specific cytotoxicity against tumor cells with positive OC166-9, which was related to the limitation of MHC. The tumor weights of SCI

关 键 词：抗独特型微抗体 卵巢肿瘤 SKOV3细胞 移植瘤 CIK 免疫细胞治疗 小鼠 

分 类 号：R737.31[医药卫生—肿瘤]