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目的用48 h 快速培养获得的成熟树突细胞与肝癌细胞系 HCCLM3构建融合细胞疫苗。方法用 CD14正选磁珠从外周血中分离出CD14~+细胞,加入含有 GM-CSF 和 IL4的树突细胞完全培养基培养24 h,再加入肿瘤坏死因子(TNF)-α、IL-1β、IL-6和 PGE2,继续培养24 h 获得树突细胞并检测免疫分子 CD80、CD86、CD83和 HLA-DR 的表达。用50%聚乙二醇+10%二甲基亚砜融合 HCCLM3与所获树突细胞构建融合细胞并检测其免疫分子的表达。自体 T 细胞增殖实验按照刺激细胞不同分为融合细胞组(RH)、树突细胞组(DC)、HCCLM3组(H)及树突细胞与 HC-CLM3混合组(HH)。结果48 h 培养获得成熟树突细胞的 CD80、CD86、CD83和 HLA-DR 表达率可达94.43%、99.71%、62.78%和99.34%,它与 HCCLM3构建的融合细胞同样表达此类免疫分子,并且对自体 T 细胞增殖的刺激作用更强(P<0.05)。结论用48 h 快速培养获得的树突细胞可以成功构建能有效刺激自体 T 细胞增殖反应的融合细胞,该方法优点显著。
Objective To construct a fusion cell vaccine with mature dendritic cells rapidly cultured in 48 h and hepatoma cell line HCCLM3. Methods CD14 ~ + cells were isolated from peripheral blood by CD14 positive magnetic beads and cultured for 24 h with complete medium of dendritic cells containing GM-CSF and IL4. Tumor necrosis factor (TNF) -α and IL-1β , IL-6 and PGE2, cultured for 24 h to obtain dendritic cells and detect the expression of CD80, CD86, CD83 and HLA-DR. Fusion of HCCLM3 with the obtained dendritic cells was established by fusion of 50% polyethylene glycol + 10% dimethylsulfoxide and the expression of their immune molecules was examined. The autologous T cell proliferation assay was divided into three groups according to the stimulated cells: fusion cell group (RH), dendritic cell group (DC), HCCLM3 group (H) and dendritic cells mixed with HC-CLM3 group (HH). Results The expression rates of CD80, CD86, CD83 and HLA-DR in mature dendritic cells after culturing for 48 h were 94.43%, 99.71%, 62.78% and 99.34%, respectively. The expression of CD80, CD86, CD83 and HLA-DR was also expressed in HCCLM3 fusion cells Stimulation of autologous T cell proliferation was stronger (P <0.05). Conclusion The dendritic cells obtained from 48 h rapid culture can successfully construct fusion cells that can effectively stimulate autologous T cell proliferation. The method has obvious advantages.