骨肉瘤细胞与活化B淋巴细胞融合疫苗的制备及其诱导的抗瘤活性

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。

Cell hybridization of a murine osteosarcoma LM9 with activated B lymphocytes for the preparation of tumor vaccines

Objective To establish osteosarcoma vaccine by the LM9 osteosarcoma derived from C3h mice hybridized with activated B lymphocytes and study its biological behavior and the antitumor efficacy. Methods The LM9 osteosarcoma derived from C3h mice was fused with LPS activated B lymphocytes by using 50%PEG. The fused cells was selected by HAT medium and cultured in vitro, and the biocharacter and efficacy of the fusion vaccine were investigated. Results In contrast to LM8, the fused cells grew significantly slowly in vitro. All the mice under the protection of fused vaccine survived without tumor (8/8), while all the mice in the control group succumbed to the tumor with no survival (8/8), 75%of the mice inoculated subcutaneously with cell fusion vaccine survived without tumor burden after implantation of LM8 cells subcutaneously. The mice in the control group developed tumors and died within 45 days without any exception. Conclusion It is possible that the LM9 osteosarcoma biology characteristics change after fused with LPS activated B lymphocytes. Cell fusion osteosarcoma vaccine could produce prophylactic and therapeutic effects in mice.