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Retrovirally Transduced CD34++ Human Cord Blood Cells
Generate T Cells Expressing High Levels of the Retroviral Encoded Green
Fluorescent Protein Marker In Vitro
Bruno Verhasselt,
Magda De Smedt,
Rita Verhelst,
Evelien Naessens, and
Jean Plum
From the Department of Clinical Chemistry, Microbiology and
Immunology, University of Ghent, University Hospital of Ghent, Ghent,
Belgium.
Human umbilical cord blood (UCB) hematopoietic stem cells (HSC)
receive increased attention as a possible target for gene-transfer in
gene therapy trials. Diseases affecting the lymphoid lineage, as
adenosine deaminase (ADA) deficiency and acquired immunodeficiency syndrome (AIDS) could be cured by gene therapy. However, the T-cell progenitor potential of these HSC after gene-transfer is largely unknown and was up to now not testable in vitro. We show here that
highly purified CD34++ Lineage marker-negative
(CD34++Lin ) UCB cells generate T,
natural killer (NK), and dendritic cells in a severe combined
immunodeficient mouse fetal thymus organ culture (FTOC).
CD34++Lin and
CD34++CD38Lin UCB cells
express the retroviral encoded marker gene Green Fluorescent Protein
(GFP) after in vitro transduction with MFG-GFP retroviral supernatant.
Transduced cells were still capable of generating T, NK, and dendritic
cells in the FTOC, all expressing high levels of GFP under control of
the Moloney murine leukemia virus (MoMuLV) long terminal repeat
promotor. We thus present an in vitro assay for thymic T-cell
development out of transduced UCB HSC, using GFP as a marker gene.
Blood, Vol. 91 No. 2 (January 15), 1998:
pp. 431-440
© 1998 by The American Society of Hematology.
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