肺癌树突状融合细胞的制备及其生物学特征
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作者：陆燕蓉 林苹 张洁 王修杰 周宏远 黄孝忠 宁奇志
单位：华西医科大学附一院肿瘤中心肿瘤研究所，四川 成都 610041
关键词：肺癌；树突状细胞；细胞融合；生物学特征
免疫学杂志010118 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。
, http://www.100md.com
分类号：Q78 文献标识码：A
文章编号：1000-8861(2001)01-0060-04
Establishment and biological characteristics of fusions of lung cancer cell with dendritic cell
LU Yan-rong(Cancer Center and Cancer Institute of the First University Hospital， West China University of Medical Science， Chengdu 610041， China)
LIN Ping(Cancer Center and Cancer Institute of the First University Hospital， West China University of Medical Science， Chengdu 610041， China)
, 百拇医药
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)
, 百拇医药
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.
, 百拇医药
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)
作者简介：陆燕蓉(1964-)，女，四川成都市人，助理研究员，硕士，主要从事肿瘤疫苗研究。 Tel：(028)5501281；E-mail： ld＠homeway.com
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.
, http://www.100md.com
[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.
, 百拇医药 [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.
收稿日期：2000年9月21日
修稿日期：2000年10月20日
出版日期：2001年1月15日, 百拇医药
单位：华西医科大学附一院肿瘤中心肿瘤研究所，四川 成都 610041
关键词：肺癌；树突状细胞；细胞融合；生物学特征
免疫学杂志010118 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。
, http://www.100md.com
分类号：Q78 文献标识码：A
文章编号：1000-8861(2001)01-0060-04
Establishment and biological characteristics of fusions of lung cancer cell with dendritic cell
LU Yan-rong(Cancer Center and Cancer Institute of the First University Hospital， West China University of Medical Science， Chengdu 610041， China)
LIN Ping(Cancer Center and Cancer Institute of the First University Hospital， West China University of Medical Science， Chengdu 610041， China)
, 百拇医药
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)
, 百拇医药
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.
, 百拇医药
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)
作者简介：陆燕蓉(1964-)，女，四川成都市人，助理研究员，硕士，主要从事肿瘤疫苗研究。 Tel：(028)5501281；E-mail： ld＠homeway.com
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.
, http://www.100md.com
[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.
, 百拇医药 [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.
收稿日期：2000年9月21日
修稿日期：2000年10月20日
出版日期：2001年1月15日, 百拇医药