[Transduction of dendritic cells by recombinant adeno-associated virus (rAAV) carrying hepatitis B virus antigens].
OBJECTIVE
To test the efficiency of infection of recombinant adeno-associated virus (rAAV) carrying hepatitis B virus S, C or X antigen, rAAV-HBV-S, C, X to human dendritic cells.


METHODS
Recombinant AAV plasmids containing HBV-S, C or X gene were constructed and packaged into rAAV in 293 cells. Monocytes were isolated from healthy donor and pulsed by rAAV-HBV-S, X, C or 293 lysate as control at the first day of isolation, then the dentritic cells were cultured for 7 days in vitro. The transcription and expression of HBV-S, C or X gene were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) or intracellular staining with fluorescence activated cell sorter (FACS), respectively.


RESULTS
The titer of rAAV-HBV-S, C, X virus was approximately 10(-7) copies per ml. After infection the HBV-S, C or X transcription expression could be seen by RT-PCR in the infected dendritic cells, the efficiency was about 90 percent by FACS.


CONCLUSIONS
rAAV-HBV-c can effectively infect and pulse dendritic cells.
You
Hong
H
Liver Center, Beijing Friendship Hospital, Capital Medical University, 100050 China. youhong30@sina.com
Cong
Min
M
Wang
Ping
P
Yan
Zhong-yu
ZY
Xu
Yong
Y
Lu
Yan
Y
Wang
Bao-en
BE
Jia
Ji-dong
JD
D002460:Cell Line; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000821:virology*; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D014161:Transduction, Genetic / Q000379:methods*
D006514:Hepatitis B Surface Antigens