肺癌树突状融合细胞的制备及其生物学特征

Establishment and biological characteristics of fusions of lung cancer cell with dendritic cell

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 L ewis肺癌细胞株 AL990 1 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 AL990 1 以 6∶ 1比例融合 ,融合率约为 30 %。融合细胞 FL D- A1 1 可在体外传代培养 ,表型为 CD80 + 、CD40 + 、H- 2 Kb + 、MIDC+ 和肺癌抗原阳性。 FL D- A1 1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 FL D- A1 1 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 FL D- A1 1 作为肿瘤疫苗 。

ObjectiveTo 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, AL 9901 , 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 AL 9901 at a ratio of 6∶1 and the fusion rate was about 30% and the resultant FLD A 11 hybrid cell was obtained through screening The FLD A 11 cell could proliferate slowly in vitro and express CD80, CD40, H 2K b, MIDC and lung cancer antigen There was no colony formed in soft agar, and no tumor mass formed in vivo after injection of FLD A 11 at 2×10 6 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 A 11 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