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目的 探讨用杂交瘤技术制备树突状细胞疫苗的方法。研究树突状细胞 肿瘤细胞之融合细胞的形态及表型特征。 方法 用PEG法将免疫磁珠法分离获得的人外周血树突状细胞与人肺鳞癌细胞株LTEP78融合 ,瑞氏 姬姆萨染色观察树突状细胞、LTEP78细胞及其融合细胞的形态结构 ,SP免疫细胞化学染色法检测融合细胞的分子表达。 结果 分离的树突状细胞能高表达HLA DR分子 ,而LTEP78细胞则不表达 ;将树突状细胞与LTEP78细胞按 10∶1比例融合后 ,两者的细胞膜、细胞质依次融合 ,获得的融合细胞仍能表达HLA DR分子。 结论人树突状细胞与人肺鳞癌细胞的融合是一个动态过程 ,通过将两者融合 ,人肺癌细胞获得了人树突状细胞表达的HLA DR分子 ,从而增强了其免疫原性 ,为人树突状细胞疫苗的研制奠定了基础。
Objective To investigate the method of preparing dendritic cell vaccine by hybridoma technology. To study the morphology and phenotypic characteristics of fused cells of dendritic cell tumor cells. Methods Human peripheral blood mononuclear cells dendritic cells isolated by immunomagnetic beads method were fused with human lung squamous cell carcinoma cell line LTEP78 by PEG method. The morphological structure of dendritic cells, LTEP78 cells and their fusion cells were observed by Wright Giemsa staining , SP immunocytochemical staining of fusion cells detected molecular expression. Results The isolated dendritic cells could express HLA DR molecules highly, but not LTEP78 cells. After dendritic cells and LTEP78 cells were fused in 10: 1 ratio, the cell membrane and cytoplasm of the two cells were fused in turn. The obtained fusion cells Still can express HLA DR molecule. Conclusions The fusion of human dendritic cells and human lung squamous cell carcinoma cells is a dynamic process. By fusion of the two, human lung cancer cells acquire HLA DR molecules expressed by human dendritic cells and enhance their immunogenicity. Dendritic cell vaccine laid the foundation for the development.