【摘要】 目的:研究Flt-3配体对树突状细胞结直肠癌细胞融合疫苗的制备和抗肿瘤免疫效应的影响。方法:①酪氨酸激酶受体3配体(FL)与粒细胞巨噬细胞集落刺激因子(GMCSF)、白细胞介素4(IL4)、肿瘤坏死因子(TNF)联用,刺激外周血单个核细胞(PBMC)中的树突状细胞(DC)前体细胞成熟,应用聚乙烯二醇(PEG)融合技术融合DC和Lovo细胞,制备融合疫苗。②通过细胞生长曲线、细胞表面表达和细胞形态来观察细胞的生物学特性。③裸鼠体内融合疫苗的成瘤活性的检测。④MTT比色法检测肿瘤细胞体外杀伤细胞毒活性。结果:在FL的参与下,成熟DC的细胞数目及细胞形态无显著改变;CD80、CD83、CD86等在细胞表面的表达较未加FL组显著丰富;融合疫苗呈悬浮生长,具有两种亲代细胞的细胞形态结构特点,融合细胞数量较稳定,不出现明显的细胞倍增;融合疫苗接种裸鼠体内后观察30d,未见肿瘤组织生长;FL组融合疫苗对肿瘤细胞具有明显的抑制作用。结论:Flt3L可显著促进DC的成熟;FL组融合细胞的分子表达较非FL组有很大提高,融合细胞具有作为抗肿瘤疫苗应具备的低增殖能力和不成瘤的安全性;DC与结直肠癌细胞融合后作为肿瘤疫苗在体外具有高效而特异的抗肿瘤效应,说明FL对增强融合疫苗抗结直肠癌特异性免疫效应有明显的作用。更多还原

【Abstract】 Objective:To investigate the effect of flt-3 ligand on the preparation of the fusional vaccine with dendritic cells and colorectal cancer cells,and the anti-tumor immunological reaction of the vaccine. Methods: ①In the presence of Flt-3 ligand、GM-CSF、IL-4 and TNF,the maturation of dendritic cells was made from peripheral blood;The preparation of the fusional vaccine was made in the presence of PEG. ②The biological characteristics of fusion cells,including their morphological specialties, the expression of cell surface phenotypes, their growth curves, inducing specific cytotoxic T lymphocyte(CTL), their possibility to proliferate into a new carcinoma in nude mice etc, were tested to verify their safety when used as anti-tumor immune reactions. Results: ①Flt-3 ligand can stimulate the full maturation of dendritic cells. Dendritic cells and Lovo cells could be fused by PEG. ②The fusion cells exhibited typical biological characteristics. They could not proliferate into new carcinomas in nude mice, and can inhibit the proliferation of Lovo cells efficiently. Conclusion: Flt-3 ligand can enhance the maturation of dendritic cell. The fused cells had poor proliferation abilities and could not proliferate into new carcinomas in immunodeficient animals. The fusion vaccine could induce specific cytotoxic T lymphocytes in vitro and was safe as a vaccine.更多还原