By the same authors

Nonviral and viral gene transfer into different subsets of human dendritic cells yield comparable efficiency of transfection

Research output: Contribution to journalArticle

Author(s)

  • A Lundqvist
  • G Noffz
  • M Pavlenko
  • S Saeboe-Larssen
  • T Fong
  • N Maitland
  • P Pisa

Department/unit(s)

Publication details

JournalJournal of immunotherapy
DatePublished - 2002
Issue number6
Volume25
Number of pages10
Pages (from-to)445-454
Original languageEnglish

Abstract

Among the many promising cancer immunotherapeutic strategies, dendritic cells (DC) have become of particular interest. This study aims to optimize a clinical grade protocol for culture and transfection of human DC. Monocytes and CD34(+) hematopoietic stem cells (HSC) from same donor were differentiated under serum-free conditions and analyzed for their susceptibility to several recently described nonviral transfection methods as compared with established virally mediated gene transfer. Nonviral gene transfer methods studied were square-wave electroporation, lipofection, and particle-mediated transfer of plasmid DNA or in vitro transcribed mRNA. We conclude that DNA is not suitable for transduction of DC using nonviral methods. In contrast, mRNA and square-wave electroporation reproducibly yields 60% and 50% transfected monocyte- and CD34(+)-derived DC, respectively, measured at protein level, without affecting the cell viability. Thus, the transfection efficiency of this method is comparable with the 40-90% transgene expression obtained using retroviral (RV) or adenoviral (AdV) vectors in CD34(+)- and monocyte-derived DC, respectively. In monocyte-derived DC, however, the amount of protein expressed per-cell basis was higher after AdV (MOI = 1000) compared with mRNA electroporation-mediated transfer. This is the first study directly demonstrating side-by-side that mRNA electroporation into DC of different origin indeed results in a comparable number of transduced cells as when using virus-mediated gene transfer.

    Research areas

  • dendritic cells, electroporation, gene transfer, transfection efficiency, CYTOTOXIC T-LYMPHOCYTES, BLOOD CD34(+) CELLS, ESTABLISHED PULMONARY METASTASES, THERAPEUTIC ANTITUMOR IMMUNITY, RESPONSES IN-VITRO, MESSENGER-RNA, CORD-BLOOD, ADENOVIRUS VECTORS, ENCODED ANTIGEN, VIRUS

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