CAJ | 학술논문

目的通过体外杀伤试验,比较4种人肝癌特异性甲胎蛋白抗原表位肽段PLFQVEPV[hAFP(137～145)],FMNKFIYEI[hAFP(158～166)]、GLSPNLNRFL[hAFP(325-334)]和GVAL,QTMKQ[hAFP(542～550)]修饰树突状细胞(DC)后诱导的细胞毒性T淋巴细胞(CTL)对人肝癌细胞的特异性杀伤效应. 方法选取健康人外周血单个核细胞,体外诱导成熟DC;合成4种hAFP抗原表位肽段分别修饰和诱导DC,体外刺激CTL,通过流式细胞仪法和细胞毒性检测法检测该修饰后的DC所诱导的CTL在体外对肝癌细胞株SMMC-7721细胞的杀伤作用.组间比较应用t检验进行统计学分析. 结果 4种人肝癌特异性甲胎蛋白片段均能在体外修饰和诱导DC细胞的增殖和成熟;这些成熟DC在体外均能诱导特异性CTL,CTL对SMMC-7721细胞均可产生杀伤效应.其中FMNKFIYEI[hAFP(158～166)]肽段诱导的CTL在效靶比分别为80:1、40:1及10:1时对肿瘤细胞的杀伤效应分别达到了78.1%±9.8%、43.9%±5.9%和28.2%±4.9%,比其他3种肽段的杀伤效应高(P<0.05). 结论短肽段具有单独体外诱导特异CTL的作用,其诱导效果和杀伤效果均优于完整hAFP蛋白,其诱导的CTL有特异地杀伤SMM C-7721细胞的作用.
목적 통과체외살상시험,비교4충인간암특이성갑태단백항원표위태단PLFQVEPV[hAFP(137～145)],FMNKFIYEI[hAFP(158～166)]、GLSPNLNRFL[hAFP(325-334)]화GVAL,QTMKQ[hAFP(542～550)]수식수돌상세포(DC)후유도적세포독성T림파세포(CTL)대인간암세포적특이성살상효응. 방법 선취건강인외주혈단개핵세포,체외유도성숙DC;합성4충hAFP항원표위태단분별수식화유도DC,체외자격CTL,통과류식세포의법화세포독성검측법검측해수식후적DC소유도적CTL재체외대간암세포주SMMC-7721세포적살상작용.조간비교응용t검험진행통계학분석. 결과 4충인간암특이성갑태단백편단균능재체외수식화유도DC세포적증식화성숙;저사성숙DC재체외균능유도특이성CTL,CTL대SMMC-7721세포균가산생살상효응.기중FMNKFIYEI[hAFP(158～166)]태단유도적CTL재효파비분별위80:1、40:1급10:1시대종류세포적살상효응분별체도료78.1%±9.8%、43.9%±5.9%화28.2%±4.9%,비기타3충태단적살상효응고(P<0.05). 결론 단태단구유단독체외유도특이CTL적작용,기유도효과화살상효과균우우완정hAFP단백,기유도적CTL유특이지살상SMM C-7721세포적작용.
Objective To compare the in vitro killing effects of cytotoxic T lymphocytes (CTLs) induced by dendritic cells(DCs) that modified with 4 different specific hAFP-derived peptides. Methods DCs derived from normal human peripheral monocytes were activated by GM-CSF and IL-4. MTT assay was applied to analyse the proliferation activities of the DCs. To induce the specific CTLs, DCs modified with four immunodominant epitopes from alpha fetoprotein (AFP) were cocultured with MNC derived CD8+ lymphocytes. The acquired specific CTLs were coclutured with SMMC-7721 cells at different dilutions, and the killing effects were detected by flow-cytometry and Non-Radiactive Cytotoxity kit. Results The CTLs stimulated by DCs modified with the four immunodominant epitopes from alpha fetoprotein (AFP) had spedific killing activities on the SMMC-7721 cells. And statistical differences of the killing effects existed between the hAFP (158-166) (FMNKFIYEI) group and the other three groups. Conclusion CTLs induced by the DCs modified with the four immunodominant peptides had significant killing effects on the hepatocellular cancer cells in vitro. The hAFP(158-166) (FMNKFIYEI) peptide had a relatively higher efficiency in inducing DCs and stimulating specific CTLs.