肺癌树突状融合细胞的制备及其生物学特征

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;对融合细胞进行生曲线、克隆形成和体内成瘤等鉴定。结果 树突状细胞与ＡＬ９９０１及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－２Ｋ＾ｂ＋、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

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

Objective To explore the established method of tumor vaccinegenerated 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.