负载食管癌抗原的DC诱导特异性CTL对食管癌细胞体外杀伤作用观察

目的观察负载食管癌抗原的树突状细胞（DC）活化的特异性细胞毒性T淋巴细胞（CTLs）对食管癌细胞的体外杀伤作用。方法冻融法获取食管癌细胞抗原,联合应用粒细胞—巨噬细胞集落刺激因子（rhGM-CSF）、白细胞介素4（IL-4）和肿瘤坏死因子α（TNF-α）诱导培养外周血DC并负载肿瘤抗原,激活自体T淋巴细胞,制备特异性CTLs。将其加入食管癌细胞中培养48 h,用MTT法检测食管癌细胞裂解率,ELISA法检测γ干扰素（γ-IFN）水平。结果负载食管癌抗原的DC激活的CTLs表现出对食管癌Eca109细胞的特异性杀伤作用,γ-IFN水平为（1 625±37.55）pg/ml;而对A549细胞仅有微弱的杀伤作用,γ-IFN水平为（169.04±13.81）pg/ml。未负载食管癌抗原DC刺激的CTLs对食管癌细胞几无杀伤作用。结论负载食管癌抗原的DC激活的CTLs在体外对食管癌细胞能产生高效而特异性的杀伤作用。

Observation of CTLs' cytotoxicity induced by DC loading esophageal antigen to esophageal cancer cell in vitro

Objective To observe CTLs′ cytotoxicity induced by DC loading esophageal antigen to esophageal cancer cell in vitro.Methods Esophageal cancer cell antigen was manufactured by rapid freezing and thawing.The DC cells were cultured with cytokines GM-CSF,IL-4,and TNF.Finally,cytotoxic T lymphocyte（CTL） activity and interferon gamma（γ-IFN） secretion was evaluated in vitro.Results CTLs induced by DC loading esophageal antigen showed cytotoxicity to esophageal cancer cell line Eca109 in vitro,the γ-IFN level was（1.625 ± 37.55 ） pg/ml ; the cytotoxicity to e-sophageal cancer cell line A549 was weak, γ-IFN level was （ 169.04 ±13.81 ）pg/ml; CTLs induced by DC without esophageal antigen showed no cytotoxicity to esophageal cancer cell line Eca109 in vitro. Conclusion CTLs induced by DC loading esophageal antigen show cytotoxicity to esophageal cancer cell in vitro.