树突状细胞与热休克处理抗原联合诱导抗肿瘤免疫的研究

目的建立体外培养与扩增脐血树突状细胞(DC)的方法,并利用DC与热休克处理的肿瘤抗原联合诱导产生一种高效特异的抗肿瘤免疫.方法用GM-CSF和IL-4联合刺激诱导脐血单个核细胞分化为DC,再用加热处理的肿瘤抗原负载,与同源的淋巴因子活化的杀伤(LAK)细胞共培养.用电镜观察DC形态,CD1a和HLA-DR试剂盒检测DC细胞表面CD1a和HLA-DR的表达情况,MTT法测定细胞杀伤活性.结果在体外,DC-LAK对同源的SPC-A-1细胞具有相对特异的杀伤作用,LAK和DC-L的作用不具有明显特异性,DC-LAK的细胞毒作用最强;加热抗原组作用明显优于未加热处理组.结论将DC与热休克处理的肿瘤抗原联合,可以诱导产生一种高效并具有相对特异性的抗肿瘤免疫活性细胞.

High potency and specific anti- tumor immunoreaction induced by dendritic cells and heat shock tumor antigen

Objective To culture dendritic cells in vitro and to induce specific anti-tumor immunoreaction in cancer cells with DC and tumor antigen.Methods DCs were cultured from umbilical cord blood mononuclear cells by using GM-CSF and IL-4,and then were stimulated by tumor antigen.DCs and lymphokine activity killer(LAK)cells or lymphocytes were cultured to induce specific anti-tumor effect,the obtained cells were named DC-LAK or DC-L.Phenotypes of DCs were analysed by reagent kit,MTT assay was used to determine cytotoxity in vitro.Results DC-LAK could lyse SPC-A-1 cells specifically in vitro.LAK and DC-L had no obvious specificity.These three effector cells can all lyse tumor cells,and the most potenct one was DCs-LAK.Conclusion High potency and relatively specific anti-tumor immunocytes can be prepared in vitro through combination of DCs and heat shock tumor antigen treatment.