CAJ | 학술논문

目的研究人外周血单核细胞来源的树突状细胞(DC)转染含甲胎蛋白(AFP,137-145)片段的重组腺相关病毒后所诱导的特异性T细胞对肝癌细胞株HepG2和SMMC-7721的体外杀伤作用.方法抽取健康志愿者外周血,分离单核细胞,体外培养,使用含AFP片断的重组腺相关病毒转染未成熟DC,诱导特异性T细胞.检测体外培养的DC和细胞毒性T细胞(CTL)活性,应用四甲基偶氮唑蓝(MTT)法检测CTL对HepG2和SMMC-7721细胞的杀伤作用.结果转染或未转染的体外培养的成熟DC高表达CD40、CD86和IL12,成熟DC诱导的CTL高表达IFNγ;修饰成熟后的DC后体外能诱导特异性CTL,该CTL在休外对肝癌细胞株HepG2和SMMC-7721均有杀伤作用.结论重组腺相关病毒转染DC,不明显改变DC表型和刺激淋巴细胞增殖和分化功能,可诱导自体CTL增殖,含AFP(137～145)片断的腺相关病毒转染DC诱导自体CTL对肝癌细胞株HepG2和SMMC-7721细胞有明显杀伤作用,DC疫苗可以作为肝癌患者免疫治疗的有效补充.
목적 연구인외주혈단핵세포래원적수돌상세포(DC)전염함갑태단백(AFP,137-145)편단적중조선상관병독후소유도적특이성T세포대간암세포주HepG2화SMMC-7721적체외살상작용.방법 추취건강지원자외주혈,분리단핵세포,체외배양,사용함AFP편단적중조선상관병독전염미성숙DC,유도특이성T세포.검측체외배양적DC화세포독성T세포(CTL)활성,응용사갑기우담서람(MTT)법검측CTL대HepG2화SMMC-7721세포적살상작용.결과 전염혹미전염적체외배양적성숙DC고표체CD40、CD86화IL12,성숙DC유도적CTL고표체IFNγ;수식성숙후적DC후체외능유도특이성CTL,해CTL재휴외대간암세포주HepG2화SMMC-7721균유살상작용.결론 중조선상관병독전염DC,불명현개변DC표형화자격림파세포증식화분화공능,가유도자체CTL증식,함AFP(137～145)편단적선상관병독전염DC유도자체CTL대간암세포주HepG2화SMMC-7721세포유명현살상작용,DC역묘가이작위간암환자면역치료적유효보충.
Objective Dendritic cell vaccines are one of the important active immunotherapies for neoplasms. The aim of this study was to observe the killing effect of specific cytotoxic T lymphocytes (CTL)on colorectal carcinoma HepG2 and SMMC-7721 cells in vitro. The CTL was induced by human peripheral blood mononuclear cells-originated dendritic cells (DC) transfected by recombinant adeno-associated virus (rAAV) with hAFP gene fragment (137-145). Methods Immature DCs were generated from peripheral blood mononuclear cells of healthy volunteers and then transfected by rAAV with AFP gene fragment. The CTL was thereafter induced. The activities of DC and CTL were measured and the killing effect of the CTL on HepC2 cells was detected using MITr assay. Results The mature DC, transfected or not, highly expressed CD40,CD86 and IL-12. IFN-γ was highly expressed in the CTL. The DC-induced CTL could effectively recognize and destroy the HepG2 and SMMC-7721 cells. Conclusion DC transfected by rAAV can stimulate the proliferation and differentiation of lymphocytes and also induce the proliferation of CTL, and their own phenotypes are not significantly altered. The DC vaccine can be effectively used as an adjuvant immunotherapy for patients with hepatocellular carcinoma.