Study on the killing effect of cytotoxic T lymphocytes induced by dendritic cells transduced recombinant adenovirus associated virus with different hepatitis B virus gene fragment
10.3969/j.issn.1671-8348.2016.27.001
- VernacularTitle:携带不同 HBV抗原基因片段的重组腺相关病毒转染树突状细胞诱导CTL的效应研究
- Author:
Fang WEI
- Publication Type:Journal Article
- Keywords:
hepatitis B virus;
dendritic cells;
T-lymphocytes,cytotoxic;
major histocompatibihity complex;
adeno-associated virus
- From:
Chongqing Medicine
2016;45(27):3745-3747,3752
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the killing effect of cytotoxic T lymphocytes(CTLs) induced by dendritic cells transduced recombinant adenovirus associated virus (rAAV ) with different hepatitis B virus gene fragment (rAAV‐HBV‐S ,C ,E ,X ) . Methods Peripheral blood mononuclear cells(PBMCs) from chronic hepatitis B patients were isolated and transduced recombinant adeno‐associated virus with different hepatitis B virus (HBV) antigen gene fragment (rAAV‐HBV‐S ,C ,E ,X) ,then GM‐CSF ,IL‐4 and TNFαwere added to cultivate for 7 days to generate mature dendritic cells (DCs) .The state of DCs were observed and differen‐tiation antigen molecules (CD)were detected by flow cytometry(FACS)to evaluate their maturation and function .Cytotoxic T lym‐phocytes (CTLs) were induced by the mixed cuture of DCs with prepared T lymphocytes .HepaG2 .2 .15 and HepaG2 were targed cells and the killing effect of CTLs wered compared using MTS assay .Results The expression of phenotype CD14 ,CD80 ,CD83 , CD86 from DCs transduced with rAAV‐HBV‐S ,C ,E ,X were compared ,respectively .Of which ,CD80 ,CD86 were significantly dif‐ferent(P<0 .05) .CD80 in group rAAV‐HBV‐X was the highest ,and CD83 in group rAAV‐HBV‐S was the highest .Cytotoxic effect of CTL induced by DC transduced with HBV antigen gene fragment of rAAV (S ,C ,E ,X) to target cells with HBV (HepG2 . 2 .15 cells) were significantly higher than that of without HBV target cells (HepG2 cells) (P<0 .01) ,but the four groups were no significant difference ,respectively(P>0 .05) .Conclusion HBV‐S ,C ,E ,X genes all induce M HC‐I dependently cytotoxic T‐lym‐phocytes specific response based adeno‐associated virus (AAV) vector delivery into dendritic cells .
