Enhancement of adenovirus-mediated gene transfer to human bone marrow cells

Tsutomu Watanabe, Linda Kelsey, Ana Ageitos, Charles Kuszynski, Kazuhiko Ino, Dean G. Heimann, Michelle T. Varney, H. Michael Shepard, Mei T. Vaillancourt, Daniel C. Maneval, James E. Talmadge

Research output: Contribution to journalReview article

13 Scopus citations

Abstract

Adenovirus infection of CD34+ hematopoietic stem/progenitor cells is dependent on the multiplicity of infection (MOI), time of incubation, the volume in which the co-incubation occurs and the presence or absence of growth factors. Studies revealed that a brief co-incubation (1-8 hours), resulted in low levels of transgene expression, suggesting that adenovirus infection of CD34+ cells occurs slowly, and optimal transduction requires a 24 hour exposure to adenovirus. Infection by Ad/β-gal or Ad/p53 at a MOI of 500:1 provided a high transduction efficiency but inhibited hematopoietic function. However, treatment at a MOI of 50-100 resulted in efficient transduction (10.7-15.7% positive) without detectable toxicity. Secondary proof of adenovirus transgene expression was demonstrated by detection of mRNA for p53 in Ad/p53 infected stem cells. We conclude that a 24 hour exposure to recombinant adenovirus encoding p53 or β-gal, at a MOI of 50-100 is optimal for in vitro gene transfer to BM cells and has no significant effect on hematopoietic function. Adenovirus-mediated transduction of BM cells can also be modulated by growth factors (IL-3, GM-CSF and G-CSF) with improved gene delivery and maintenance of hematopoietic function. In summary, adenovirus vectors can be used to transiently transduce stem cells, and conditions have been defined to maximize expression and limit inhibitory effects on CD34+ cells. These data support continued investigation of this vector for local cytokine delivery and purging of stem cell products.

Original languageEnglish (US)
Pages (from-to)439-451
Number of pages13
JournalLeukemia and Lymphoma
Volume29
Issue number5-6
DOIs
StatePublished - 1998

Keywords

  • Adenovirus
  • CD34 cells
  • Gene therapy
  • Hematopoiesis
  • Purging

ASJC Scopus subject areas

  • Hematology
  • Oncology
  • Cancer Research

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