脐血树突状细胞介导的食管癌疫苗抗肿瘤实验研究

The antitumor effect of cancer vaccine prepared by fusions of esophageal carcinoma cell line with dendritic cell derived from cord blood

目的:用杂交瘤技术制备脐血树突状细胞(DC)和食管癌细胞的融合疫苗,观察其在体内外抗肿瘤的作用。方法:分离脐血CD34+干细胞并诱导扩增为成熟DC,并与EC109细胞经聚乙二醇法融合;免疫磁珠法筛选EC109-DC;MTT法测定瘤苗诱导淋巴细胞增殖能力及体外特异性免疫应答能力;重建SCID小鼠免疫功能;体内抗肿瘤免疫治疗实验。结果:(1)融合疫苗可体外生长,且同时高表达叶酸受体和CD80。(2)疫苗能有效刺激淋巴细胞增殖反应,体外诱导细胞毒T细胞(CTL)对EC109的杀伤活性显著高于对照组(P<0.05)。(3)在免疫治疗中,用融合瘤苗治疗的荷瘤小鼠,肿瘤大小和重量明显小于对照组(P<0.05)。结论:(1)EC109-DC能在体外刺激淋巴细胞增殖和诱导特异性CTL杀伤作用;(2)融合瘤苗治疗荷瘤小鼠有一定的效应。

Objective To generate a fusion vaccine of esophageal carcinoma cell and dendritic cell （DC） and study its antitumor effect. Methods Stem cells CD34^＋ were isolated from cord blood, then induced to mature DCs, and finally fused with EC109 cells. The generated fusion vaccine EC109-DC was screened by magnetic microbead marker. The capacity of proliferation and in vitro specific immune response of the lymphocytes induced by the fusion vaccine was detected by MTT The immune function of SCID mice was reconstructed and the protective effect of the fusion vaccine against EC109 was determined. Results EC109-DC could proliferate slowly in vitro with high expressions of FR and CD80, The proliferative reaction of the lymphocytes and the specific cytotoxicity against EC109 induced by the fusion vaccine were significantly higher than those in the control group （P 〈 0.05）, The mouse tumor size and tumor weight were markedly smaller in the vaccine treatment group than in the control group. Conclusions EC109-DC vaccine can stimulate lymphocyte proliferation in vitro and induce CTL activity. The fusion vaccine has certain effect on inhibiting mouse tumor.