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

S. Ponnazhagan, G. Mahendra, D. T. Curiel, D. R. Shaw

Research output: Contribution to journalArticle

69 Scopus citations

Abstract

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

Original languageEnglish
Pages (from-to)9493-9501
Number of pages9
JournalJournal of virology
Volume75
Issue number19
DOIs
StatePublished - Sep 27 2001
Externally publishedYes

Fingerprint Dive into the research topics of 'Adeno-associated virus type 2-mediated transduction of human monocyte-derived dendritic cells: Implications for ex vivo immunotherapy'. Together they form a unique fingerprint.

  • Cite this