Transduction of dendritic cells by recombinant adeno-associated virus (rAAV) carrying hepatitis B virus antigens / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>rAAV-HBV-c can effectively infect and pulse dendritic cells.</p>