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目的 :构建分泌独特型免疫球蛋白 (idiotype ,Id)的BALB c小鼠B细胞淋巴瘤细胞株 ,建立动物模型 ,为进一步开展Id 树突状细胞 (dendriticcell,DC)疫苗的免疫治疗提供物质基础。方法 :采用降植烷和不完全弗氏佐剂间隔致敏 6～ 8周龄BALB c小鼠 ,继而采用细胞融合技术、ELISA法、免疫荧光标记和流式细胞术 (flowcytometry ,FCM)筛选获得分泌Id的阳性克隆 ;体内诱生腹水和体外无血清培养法制备Id ,优球蛋白沉淀法结合凝胶Suphecray 2 0 0过滤纯化Id ;建立荷瘤模型 ,从荷瘤小鼠骨髓前体细胞诱导DC。结果 :获得 2株稳定分泌Id的BALB c小鼠B细胞淋巴瘤细胞SB4和SB5 ,Id均为IgM κ ;成瘤率均为 10 0 % ,荷瘤小鼠平均生存期为 (3 0± 4)d。结论 :SB4和SB5能持续稳定分泌非自身反应性Id ,具有良好的成瘤性和高转移率
OBJECTIVE: To construct a BALB / c mouse B cell lymphoma cell line secreting idiotype (Id) and establish an animal model to provide a material basis for further immunotherapy of Id dendritic cell (DC) vaccine . Methods: Six to eight-week-old BALB mice were sensitized with pristane and incomplete Freund’s adjuvant, and then were screened by ELISA, immunofluorescence and flow cytometry (FCM) Positive clones secreting Id; in vivo induced ascites and serum-free culture in vitro preparation Id, euglobulin precipitation with gel Suphecray 200 filter purified Id; established tumor-bearing model of tumor-bearing mouse bone marrow precursor cells induced DC. Results: Two BALB / c mice B-cell lymphoma cells SB4 and SB5 with stable secretion of Id were obtained, Id were both IgM κ; the tumorigenic rates were 100%, and the average survival time of tumor-bearing mice was (30 ± 4) d. Conclusion: SB4 and SB5 can stably secrete non-self-reactive Id, with good tumorigenicity and high metastatic rate