Adeno-associated virus type 2-mediated transduction of human monocyte-derived dendritic cells: implications for ex vivo immunotherapy.

摘要: Dendritic cells (DCs) are pivotal antigen-presenting for regulating immune responses. A major focus of contemporary vaccine research is the genetic modification DCs to express antigens or immunomodulatory molecules, utilizing a variety viral and nonviral vectors, induce antigen-specific responses that ameliorate disease states as diverse malignancy, infection, autoimmunity, allergy. The present study has evaluated adeno-associated virus (AAV) type 2 vector ex vivo gene transfer human peripheral blood monocyte (MO)-derived DCs. AAV nonpathogenic parvovirus infects wide cell lineages in vitro, long-term transgene expression without requirements proliferation. presented data demonstrate recombinant (rAAV) can efficiently transduce MOs well generated by MO culture with granulocyte-macrophage colony-stimulating factor plus interleukin vitro. rAAV MO-derived could be enhanced etoposide, previously reported enhance expression. transduction freshly purified followed 7 days cytokines generate DCs, subsequent sorting coexpression DC markers CD1a CD40, showed robust evidence nuclear localization genome population. Phenotypic analyses using multiple functional assays one-way allogeneic mixed leukocyte reactions indicated rAAV-transduced were equivalent nontransduced These results support utility vectors future studies.