肺癌树突状融合细胞的制备及其生物学特征

摘要： 探讨利用杂交瘤技术制备肿瘤疫苗的方法,研究肺癌树突状融合细胞的生物学特征.  

摘　要： 目的　探讨利用杂交瘤技术制备 肿瘤 疫苗 的方法,研究 肺癌 树突状融合细胞的生物学特征.方法　　用PEG法将HGPRT缺陷型Lewis肺 癌细胞 株AL9901与小鼠 骨髓 诱生的树突状细胞进行融合,HAT筛选融合克隆；S-P 免疫细胞 化学法鉴定融合细胞表型；对融合细胞进行生长曲线、 克隆 形成和体内成瘤等鉴定.结果　树突状细胞与AL9901以6∶1比例融合,融合率约为30％.融合细胞FLD-A11可在体外传代培养,表型为CD80＋、CD40＋、H-2Kb＋、MIDC＋和肺癌抗原阳性.FLD-A11不能在软琼脂培养基中形成克隆,动物体内不能形成肿瘤.结论　利用杂交瘤技术制备的FLD-A11细胞,具备了在体内激活抗特异性地抗 肿瘤细胞 免疫功能的分子表型；该细胞不存在体内成瘤的危险.本研究为FLD-A11作为肿瘤疫苗,用于特异性抗肿瘤免疫活性和主动免疫治疗的研究打下了基础. 关键词 ：肺癌；树突状细胞；细胞融合；生物学特征 　　Establishment and biological characteristics of fusions of lung cancer cell with dendritic cellLU Yan-rong（Cancer Center and Cancer Institute of the First University Hospital, West China University of Medical　Science, Chengdu610041, China） 　　LIN Ping（Cancer Center and Cancer Institute of the First University Hospital, West China University of Medical　Science, Chengdu 610041, China） 　　ZHANG Jie（Cancer Center and Cancer Institute of the First University Hospital, West China University of Medical　Science, Chengdu 610041, China） 　　WANG Xiu-jie（Cancer Center and Cancer Institute of the First University Hospital, West China University of Medical　Science, Chengdu 610041, China） 　　ZHOU Hong-yuan（Cancer Center and Cancer Institute of the First University Hospital, West China University of Medical　Science, Chengdu 610041, China） 　　HUANG Xiao-zhong（Cancer Center and Cancer Institute of the First University Hospital, West China University of Medical　Science, Chengdu 610041, China） 　　NING Qi-zhi（Cancer Center and Cancer Institute of the First University Hospital, West China University of Medical　Science, Chengdu 610041, China） 　　Abstract：Objective　To explore the established method of tumor vaccine generated by hybrid technique and study the biological characteristics of lung cancer-dendritic fusion cell. Methods　 HGPRT defective Lewis lung cancer cell line, AL9901, was fused with bone marrow derived dendritic cells （DC） by PEG-1000. The phenotypes and proliferation characteristic of the fused cell were observed in vitro, and its tumor generating rate was observed in vivo.Results　 The DC was fused with AL9901 at a ratio of 6∶1 and the fusion rate was about 30％ and the resultant FLD-A11 hybrid cell was obtained through screening. The FLD-A11 cell could proliferate slowly in vitro and express CD80, CD40, H-2Kb, MIDC and lung cancer antigen. There was no colony formed in soft agar, and no tumor mass formed in vivo after injection of FLD-A11 at 2×106 per mice.Conclusions　 Lung cancer vaccine could be obtained through tumor cell fusion with DC, thereby acquired the immuno-stimulating phenotype to stimulate the specific antitumor activity in vivo. The FLD-A11 cell could be applied without danger of tumor generation. These data found a basis for the study of specific antitumor immunity and active immunotherapy of DC-based vaccine. 　　Keywords：lung cancer； dendritic cell； cell fusion； biological characteristics基金项目：国家自然科学基金资助项目（39800147） 　　参考文献： 　［1］Zitvogel L, Mayordomo JI, Tjandrawan T, et al. Therapy of murine tumor with tumor　peptide-pulsed dendritic cells： dependence on T cell, B7 co-stimulation, and T help cell 1-associated cytokines［J］. J Exp Med, 1996, 183（1）：87－97. 　［2］陆燕蓉,林　苹,张　洁,等. HGPRT缺陷型Lewis肺癌 细胞株 的筛选及其生物学特征［J］.中国肺癌杂志,2000（在排印中）. 　［3］汪　惠,赖百塘,湛秀萍,等. 两个肺腺癌细胞学的建立及其生物学特性的观察［J］. 中华肿瘤杂志,1983, 5（2）：85－88. 　［4］李明松,袁爱力,潘德寿,等. 树突状细胞诱导的抗肿瘤免疫预防肝转移［J］. 实用肿瘤杂志,2000, 15（2）：115－117. 　［5］Guo Y, Wu M, Chen H, et al. Effective tumor vaccine generated by fusion of hepatoma　cells with activated B cells［J］. Science, 1994, 263（28）： 518－520. 　［6］Cao X, Zhang W, Wang J, et al. Therapy of established tumor with a hybrid cellular vaccine generated by using granulocyte-macrophage colony-stimulating factor genetically modified dendritic cells［J］. Immunology, 1999, 97（4）： 616－625. 　［7］林　苹,张　洁,陆燕蓉,等. 表达B7分子的肺癌融合细胞刺激的主动 免疫应答 ［J］.中国肺癌杂志,2000（在排印中）. 　［8］Costello RT, Gastaut JA, Olive D. What is the real role of CD40 in cancer immunotherapy?［J］ Immuno-logy Today, 1999, 20（11）： 188－192. 相关阅读： 代谢蛋白PKM2对肿瘤细胞有着意想不到的作用 研究发现抑制癌细胞扩散的关键蛋白 克隆人类 胚胎 干细胞 的新进展 肺癌靶向药物疗效与基因 突变 量有关 糖尿病 肺癌患者寿命长于普通肺癌患者