树突状细胞负载人工合成甲胎蛋白表位肽后诱导的细胞毒作用  

Specific Lysis of CTLs Induced by Dendritic Cells-added to HLA-A2 Restricted Epitopes Derived from α-fetoprotein against HCC in Vitro

作　　者：张兵[1] 董薇[1] 朱月明[1] 蔡美英[1] 

机构地区：[1]四川大学华西医学中心基础医学与法医学院免疫学教研室,四川成都610041

出　　处：《航天医学与医学工程》2007年第2期102-105,共4页Space Medicine & Medical Engineering

基　　金：国家自然科学基金项目(30070855)

摘　　要：目的观察HLA-A2树突状细胞(dendritic cells,DCs)负载人甲胎蛋白(humanα-fetoprotein,hAFP)HLA-A2限制性九肽(FMNKFIYEI,AFP158-166)后,激活特异的细胞毒性T淋巴细胞(cytotoxic Tlymphocytes,CTLs),杀伤肝癌细胞株的能力。方法用贴壁法分离HLA-A2成人外周血单核细胞,分次加入IL-4与GM-CSF共同诱导培养6d,给予单核细胞调控培养液(monocyte conditioned medium,MCM)诱导成熟2d,然后负载AFP158-166,激活特异性CTLs,用MTT法检测CTL杀伤肝细胞癌(hepa-tocellular carcinoma,HCC)细胞株HepG2和T2细胞株的能力。结果采用贴壁法、多次加入细胞因子和MCM为成熟诱导剂的培养方法可以获得足量的成熟DCs,并且在去除细胞因子后,仍能保持成熟的DCs形态;负载AFP158-166的DCs激活的CTLs对HepG2和负载AFP158-166的T2具有较强的杀伤作用。结论负载HLA-A2限制性表位肽AFP158-166的DCs对表达AFP的HLA-A2HCC有潜在的治疗作用,具有一定的临床运用前景。Objective To observe specific lysis of cytotoxic T lyphocytes （CTLs） induced by dendrictic cells （DCs）-added to HLA-A2 restricted epitopes derived from human α-fetoprotein （FMNKFIYEI , hAFP158-166） against T2 cell lines and hepatocellular carcinoma cell （ HCC ） HepG2. Methods CD14^＋ DC precursors were enriched with human peripheral blood mononuclear cells （ PBMCs ） by a modified adherence steps, then GM-CSF and IL-4 was added to adherent fraction , and on the 3rd and 5th day from beginning of culture, two kinds of cytokine were added again. On the 6th day, all nonadherent cells were harvested, then monocyte conditioned medium （ MCM ） was added to them. On the 8th day, the mature DCs were harvested, and led with AFP158-166 to activate CTLs, Lastly, specific lysis of CTLs against T2 and HepG2 was analysed by MTT. Results The more typical and mature DCs were abtained by an improved method of culture and the mature DCs remained stable and viable even if cultured for another 3 d in absence of this two cytokines. The CTL activated by the DCs-added to AFP158-166 can specificly strongly lysed the HepG2 and T2 cell lines led with the same epitopes, but slightly lysed the epitopes-free T2. Conclusion The DC-added with HLA-A2 restricted epitopes derived from hAFP can induce in vitro cytotoxicity against the HepG2 , it suggests the possibility that DCs can serve as a new type of vaccine for HCC.

关 键 词：树突状细胞 甲胎蛋白 肝细胞癌 限制性表位肽 

分 类 号：R392.32[医药卫生—免疫学] Q789[医药卫生—基础医学]