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.
Conclusion: rAAV-HBV-c can effectively infect and pulse dendritic cells.