Adenovirus mediated alpha interferon (IFN-alpha) gene transfer into CD34+ cells and CML mononuclear cells

Stem Cells. 1997;15(5):386-95. doi: 10.1002/stem.150386.


Gene transfer or gene therapy has advantages in the treatment of a variety of disorders due to its selective expression within specific mammalian cells. Interferon-alpha (IFN-alpha) has been used in the management of leukemia but its diverse adverse activities with multiple potential side effects, possibly unrelated to therapeutic targets, may negatively influence the ability of IFN-alpha to treat this disorder. Therefore, we examined the ability of adenovirus (Ad)-IFN-alpha gene construct to transfect normal (CD34+ cells) and chronic myelogenous leukemia (CML) bone marrow mononuclear cells (BMMNC) and the transient overexpression of IFN-alpha in these cells. Ad-cytomegalovirus promoter driven IFN-alpha (AdCMV-IFN-alpha) at multiple doses was assessed to transfect highly purified CD34+ cells in liquid culture, and optimal transduction of CD34+ cells was achieved using 120 plaque forming units. Flow cytometric determinations revealed that there was no significant difference in cell viability for the 4 h or 24 h transfection periods. Immunoassay of IFN-alpha produced by CD34+ cells shows that IFN-alpha levels increased several fold in transfected cells. Transient expression of the IFN-alpha gene did not suppress proliferation of CD34+ progenitors as indicated by BFU-E or colony forming units-granulocyte-macrophage (CFU-GM) growth. Reverse transcriptase/polymerase chain reaction analysis of RNA from CD34+ harvested CFU-GM progenitor cells demonstrated transient IFN-alpha mRNA expression. Similarly, CML BMMNC were transfected with AdCMV-IFN-alpha under similar conditions as described for CD34+ cells. BMMNC cells exposed to adenovirus for 24 h and 48 h were found to express IFN-alpha at a substantial level. This in vitro data suggest that Ad-mediated gene transfer of IFN-alpha into hematopoietic stem cells can be achieved and that the IFN-alpha gene can be translated into its specific mRNA in CD34 progenitor cells.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae / genetics
  • Antigens, CD34 / genetics*
  • Bone Marrow Cells / cytology
  • Colony-Forming Units Assay
  • Cytomegalovirus / genetics
  • DNA, Complementary / analysis
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Dose-Response Relationship, Drug
  • Gene Transfer Techniques*
  • Genetic Vectors*
  • Humans
  • Interferon-alpha / genetics*
  • Interferon-alpha / therapeutic use
  • Interferon-alpha / toxicity
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / therapy
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / physiology*
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Stem Cells / drug effects
  • Stem Cells / immunology
  • Stem Cells / physiology
  • Transfection


  • Antigens, CD34
  • DNA, Complementary
  • Interferon-alpha
  • RNA, Messenger