目的骨肉瘤细胞与活化B淋巴细胞融合制备肿瘤疫苗,探讨其生物学特征及抗肿瘤特性。方法以体积分数50%的聚乙二醇(PEG)为融合剂,将C3h小鼠来源的次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)基因缺陷型骨肉瘤细胞LM9与脂多糖(LPS)活化的C3h小鼠B淋巴细胞进行融合。经HAT(hypoxanthine-aminopyerin-thymidine)选择性培养基筛选后,流式细胞仪检测融合细胞表面H-2Kb表达情况,用有限稀释法进行克隆,H-2Kb高表达株FLM9用于体内抗瘤实验,观察该融合瘤株的体内外生长特性、对C3h小鼠的致瘤性以及抗肿瘤免疫活性。结果C3h小鼠骨肉瘤融合细胞在体外生长缓慢,对C3h小鼠无致瘤性,经有限稀释法克隆得到一个B7表达为75.11%的克隆,该克隆H-2Kb表达为15.6%;作用于荷瘤C3h小鼠,融合细胞治疗组小鼠(8只)2只分别于38d和45d死亡,其余6只存活期超过60d;对照组小鼠均在60d内全部死亡(P<0.01)。用肿瘤细胞攻击时,预先接种融合瘤苗组的存活率75%明显高于对照组。结论活化B淋巴细胞与骨肉瘤细胞融合后改变了肿瘤细胞的生物学特性,对小鼠有较好的预防和治疗肿瘤作用,说明用细胞融合方法构建的肿瘤疫苗具有潜在的应用价值。 Target osteosarcoma cells were fused with activated B lymphocytes to prepare tumor vaccines, and their biological characteristics and anti-tumor properties were explored. Methods The hypothine guanine phosphoribosyltransferase (HGPRT) gene-deficient osteosarcoma cell LM9 and lipopolysaccharide (LPS) derived from C3h mice were activated with a 50% volume fraction of polyethylene glycol (PEG) as a fusion agent. The C3h mouse B lymphocytes were fused. After selection by HAT (hypoxanthine-aminopyerin-thymidine) selective medium, the expression of H-2Kb on the surface of fused cells was detected by flow cytometry, and cloned by limiting dilution method. FLK9 high expression strain H2Kb was used for in vivo antitumor experiments. The in vitro and in vivo growth characteristics of the hybridoma strain, the tumorigenicity and antitumor immunity of the C3h mouse were observed. Results The C3h mouse osteosarcoma fusion cells grew slowly in vitro and had no tumorigenicity to C3h mice. A clone with a B7 expression of 75.11% was cloned by limiting dilution method. The clone H-2Kb expression was 15.6%. In tumor C3h mice, 2 mice in the fusion cell treatment group (8) died at 38 days and 45 days respectively, and the remaining 6 survived for more than 60 days; mice in the control group all died within 60 days (P<0.01). When challenged with tumor cells, the survival rate of the preinoculated fusion tumor group was 75% significantly higher than that of the control group. Conclusion The fusion of activated B lymphocytes with osteosarcoma cells alters the biological characteristics of tumor cells, and has a better effect on the prevention and treatment of tumors in mice. This shows that tumor vaccines constructed using cell fusion methods have potential applications.