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Blood, Vol. 96 No. 3 (August 1), 2000:
pp. 885-893
Highly efficient gene transfer in naive human T cells with a
murine leukemia virus-based vector
Valérie Dardalhon,
Sara Jaleco,
Cosette Rebouissou,
Christophe Ferrand,
Nadia Skander,
Louise Swainson,
Pierre Tiberghien,
Hergen Spits,
Nelly Noraz, and
Naomi Taylor
From the Institut de Génétique Moléculaire de
Montpellier, CNRS UMR 5535 (IFR24), Montpellier, France; Laboratoire de
Thérapeutique Immuno-Moléculaire, EFS Bourgogne-France
Comté, Besançon, France; Division of Immunology, The
Netherlands Cancer Institute, Amsterdam, The Netherlands.
Retroviral vectors based on the Moloney murine leukemia virus (MuLV)
have become the primary tool for gene delivery into hematopoietic cells, but clinical trials have been hampered by low transduction efficiencies. Recently, we and others have shown that gene transfer of
MuLV-based vectors into T cells can be significantly augmented using a
fibronectin-facilitated protocol. Nevertheless, the relative abilities
of naive (CD45RA+) and memory (CD45RO+)
lymphocyte subsets to be transduced has not been assessed. Although naive T cells demonstrate a restricted cytokine profile following antigen stimulation and a decreased susceptibility to infection with
human immunodeficiency virus, it was not clear whether they could be
efficiently infected with a MuLV vector. This study describes conditions that permitted gene transfer of an enhanced green
fluorescent protein-expressing retroviral vector in more than 50%
of naive umbilical cord (UC) blood and peripheral blood (PB) T cells
following CD3/CD28 ligation. Moreover, treatment of naive T cells with
interleukin-7 resulted in the maintenance of a CD45RA phenotype and
gene transfer levels approached 20%. Finally, it was determined that
parameters for optimal transduction of CD45RA+ T cells
isolated from PB and UC blood differed: transduction of the UC cells
was significantly increased by the presence of autologous mononuclear
cells (24.5% versus 56.5%). Because naive T cells harbor a receptor
repertoire that allows them to respond to novel antigens, the
development of protocols targeting their transduction is crucial for
gene therapy applications. This approach will also allow the functions
of exogenous genes to be evaluated in primary nontransformed naive T cells.
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