摘要
目的体外诱导小鼠骨髓单个核细胞分化为树突细胞(DC),并以骨髓来源的树突细胞(BM-DC)为载体研制骨髓瘤疫苗。方法在粒细胞-巨噬细胞集落刺激因子(GM-CSF)、IL-4的作用下诱导小鼠骨髓单个核细胞生成DC。利用聚二乙醇(PEG)作为融合剂,将BM-DC与SP2/0骨髓瘤细胞进行化学融合,并用含次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷(HAT)的选择性培养基对其进行筛选。观察融合细胞的生长特性、并检测其体外特异性抗肿瘤活性。结果在细胞因子的作用下培养7d并经磷酸脂多糖(LPS)刺激成熟的DC表面显示出明显的树枝状突起,高表达CD11c、CD86及主要组织相容性复合体(MHC)-Ⅱ类分子,并且在混合淋巴细胞反应中展示出很强的刺激初始T淋巴细胞增殖的能力。通过PEG融合及HAT筛选后,杂交瘤细胞在体外可诱导T细胞生成针对SP2/0骨髓瘤细胞的细胞毒性T淋巴细胞(CTL),并对其有强大的特异性杀伤作用。结论BM-DC与肿瘤细胞融合后,可激发机体产生特异性的抗肿瘤免疫反应,利用BM-DC制备抗肿瘤疫苗具有实际应用的可行性。
Objective To induce mice bone marrow （BM） mono - nuclear cells into dendritic cells （DC） in vitro, and then produce function vaccine of anti - myeloma by bone marrow derived dendritic cells （ BM - DC）. Methods Mono - nuclear cells separated from mice bone marrow were cultured in tissue culture plastics which supplemented granulocyte -macrophage clonystimulating factor （GM- CSF）, IL-4. During the 8 days, there were numerous of mature DCs outgrown, and then fused with SP 2/0 myeloma cells in logarithmic growth phase cultured with 8 - azaguaine by polyethylene （PEG）. The resulting purified DC/myeloma hybrids were generated by selecting with hypoxanthine - aminopterin - thymidine （HAT）, The specific anti - tumor activity was examined in vitro, Results Mono - nuclear cells derived from BM cultured by supplement propriate cytokines milieu in tissue culture plastics, and then exposed to lipopolysaccharide （LDS）. There were plenty of mature DCs appear as early as day 8, and exhibit dramatic veils of eytoplasm and extensive dendrites in their surfaces, high express CD11c （ 83.19% ）, major histocompatibility complex （MHC） - Ⅱ （95.25%）, CD86 （ 89. 24% ）, their capacity to stimulate primary T cell responses in mixed leukocyte reaction （MLR） would be shown strongly, Hybrids could grow well through PEG fusion and HAT selection. These hybrids could stimulate naive T cells into cytotoxic T- lymphocyte（CTLs） directed against SP 2/0 myeloma, and exhibit strong specific killed ability. Conclusions BM- DCs is fused with tumor cells ensure the tumor associated antigens could be processed and presented effectively, and stimulate specific anti -tumor immunity so that prevent or cure this kind of tumor, So it＇s feasible in practice to produce anti -tumor vaccine by BM - DCs,
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2008年第3期176-178,共3页
Journal of Applied Clinical Pediatrics
基金
教育部新世纪优秀人才支持计划项目资助(NCET-05-0820)
关键词
骨髓
树突细胞
骨髓瘤
疫苗
细胞融合
bone marrow
dendritic cell
myeloma
vaccine
cell fusion
