合成多肽体外诱导乙肝病毒抗原特异性T杀伤细胞实验研究  

IN VITRO INDUCTION OF HEPATITIS B VIRUS ANTIEGN-SPECIFIC T CELLS BY COSTIMULATION WITH SYNTHETIC PEPTIDES AND TRANSFERRIN AND CD_3mAb

作　　者：邓涛[1] 陈乃玲[1] 武珊珊 陈天宝[1] 鲁芳芳 张晓红 刘倩[1] 贾克明[1] 

机构地区：[1]中国人民解放军肝病研究所,北京100700

出　　处：《中华肝脏病杂志》1997年第1期44-46,共3页Chinese Journal of Hepatology

摘　　要：目的及方法 为探讨ＨＢＶ抗原特异性细胞毒Ｔ细胞（ＨＢＶ－ＣＴＬ），根据ＨＢｃＡｇ的ＨＬＡ分子结合关键序列合成抗原多肽２段，分别与转铁蛋白（Ｔｆ），抗ＣＤ单克隆抗体（ＣＤ３ｍＡｂ）和ＩＬ－２联合体外诱导ＨＢＶ－ＣＴＬ，应用＾３Ｈ－ＴｄＲ释放法分别测定ＨＢＶ－ＣＴＬ对ＨＢＶＤＮＡ转染的ＨｅｐＧ２．２．１５细胞特异性活性及无ＨＢＶＤＮＡ转染的ＨｅＰＧ２细胞的非特异性杀伤率。结果Objective and Methods In order to study the feasibility of induction of HBV antigen-specific T cells in vitro,the PBMC isolated from health donors'blood was costimulated with both CD3mAb and Tf and the synthetic peptide of HBcAg(peptide Ⅰ:amino acid 18-27 and peptid Ⅱ:aa 141-151).The HBV-DNA transfected hepatoma cells(2.2.15).was used as target cells for detecting the specific cytotoxicity of HBV-specific CTLs by ~3H-TdR release assay.Meanwhile, hepatoma cells(HepG2)were used for detecting the nonspecific cytotoxicity.The phenotype were analysed through APAAP staining method.Results The specific lysis aginst 2.2.15 cells were 67%(peptide Ⅰ+CD3mAb+Tf)and 52% (peptide Ⅱ+CD3mAb+Tf)respectively,which were significant higher than the nonspecific cytotoxicity aginst HepG2 25% and 30%(P<0.05).But the cytotoxicity of CTLs,stimulated only with CD3mAb and Tf,against 2.2.15 and HepG2 cells were 34% and 25%,no signifant differance was found between them.The number of HBV-specific CTLs expanded to 8- fold in 14 days'culture and 20-fold in 21 days'culture,10 times higher than LAK cells.The population of HBV-specific CTLs were CD4+20%,CD8+56%,CD71+28%.Conclusion The results suggested that the HBV-specific CTLs could be induced by costimilation with short synthetic peptides in vitro.This results also imply that the peptides derived from HBcAg(aa:18-27 and 141-151)may be the CTLs epitope.

关 键 词：合成多肽 乙型肝炎病毒 细胞毒T细胞 诱导 

分 类 号：R512.620.2[医药卫生—内科学]