小鼠骨髓来源树突状细胞与NS_1骨髓瘤细胞的融合瘤苗研制  

An effective cancer vaccine developed by fusion of NS 1 myeloma cells with the bone marrow derived dendritic cells

作　　者：张义国[1,2] 董子明[1,2] 杨洪艳[1,2] 郑智敏[1,2] 赵明耀[1,2] 吴皓[1,2] 

机构地区：[1]河南医科大学病理生理学教研室 [2]郑州铁路中心医院

出　　处：《河南医科大学学报》1999年第1期26-30,共5页Journal of Henan Medical University

基　　金：河南医科大学回国人员科研启动基金;河南医科大学基础医学院科研基金;河南省教委重点攻关项目;河南省科委重点攻关项目

摘　　要：目的：为了提高肿瘤的免疫原性，有效地引发并增强宿主体内抗肿瘤免疫应答，特研制小鼠骨髓树突状细胞（ｄｅｎｄｒｉｔｉｃｃｅｌｓ，ＤＣｓ）与ＮＳ１骨髓瘤细胞的融合瘤苗。方法：利用特异的ＣＤ４、ＣＤ８、Ｂ２２０单克隆抗体和补体缓冲液及ＤＣｓ的半粘附性，直接从骨髓细胞中分离出高纯度的ＤＣｓ及其前体；再联用粒细胞巨噬细胞集落刺激因子（ＧＭＣＳＦ）和白细胞介素（ＩＬ）４体外培育，协同诱导ＤＣｓ及其前体分化增殖，以获得大量高纯度的ＤＣｓ；然后用聚乙二醇（ＰＥＧ），使其与８氮杂鸟嘌呤（ＡＧ）处理过且处对数生长期的ＮＳ１骨髓瘤细胞融合，并用次黄嘌呤氨基蝶呤胸腺嘧啶核苷（ＨＡＴ）选择培养基筛选融合细胞，观察融合细胞的生长特性和体内致瘤性；最后直接从ＢＡＬＢ／Ｃ小鼠尾静脉免疫接种活的融合细胞，再观察其诱导宿主抗肿瘤的免疫效果。结果：①融合细胞也有刺激混合淋巴细胞增殖反应的能力，其在体外能分裂增殖，但明显低于肿瘤细胞，无体内致瘤性；②接种活融合细胞的免疫小鼠能抵抗野生肿瘤攻击长达９０ｄ未见诱发肿瘤；③对照组小鼠１００％出现诱发肿瘤。结论：融合细胞可能加工处理并表达尚未鉴定的肿瘤特异抗原，进而激发宿主体内抗肿瘤免疫反应，使?Aims:For improving the immunogenity of cancer and effectively inducing host in vivo antitumor immune responses,a novel cancer vaccine based on fusion of the bone marrow derived dendritic cells (DCs) with NS 1 myeloma cells was developed in our study.Methods: By a combination of freshly isolated procedure with anti CD4, CD8 and B220 monoclonal antibodies and complements and by semi adhesion to culture plates,and cytokine induced exponsion culture with granulocyto macrophages colony stimulating factor (GM CSF) and interleukin(IL 4),a large number of DCs with over 95% of high purity were developed,and then by 50% polyethylene glycol (PEG) fused with NS 1 myeloma cells in logarithmic growth phase cultured with 8 azaguaine (AG).The resulting hybrids were generated by selecting with hypoxanthine aminopterin thymidine (HAT) plus GM CSF and IL 4,and their characteristics of in vitro growth and in vivo tumorigenicity were analysed.Directly with freshly fused hybrids as the immunogen BALB/C mice were vaccinated intravenously,and then challeged with wild live parental myeloma cells.It was finally investigated whether the immunized animals would develop host in vivo antitumor activity induced by the fused cell vaccine.Results:This experimental study indicated that: ①The fused cells also had the potentiality of stimulating proliferation of mixed T lymphocytes and formed cell clusters with them; ②These cells could divide and proliferate,but which significantly decreased as compared with parental tumor cells;③Over 60 days after subcutaneous implantment of fusion cells was observed no induced tumorigenesis,but in controls of tumor cells the rate of tumorigenesis was 100% and the time of tumorigenicity and survival respectively was 6.4 and 13.4 days;④Fusion cell immunized animals could protect against the challege of tumor for over 90 days,but control groups with irradiated tumor cells or PBS had tumorigensis at the rate of 100% for the time respectively of 15.2 and 6.8 days.Concl

关 键 词：肿瘤疫苗 树突状细胞 骨髓肿瘤 细胞融合 小鼠 

分 类 号：R733.3[医药卫生—肿瘤] R730.3[医药卫生—临床医学]