High-efficiency gene transfer into normal and adenosine deaminase- deficient T lymphocytes is mediated by transduction on recombinant fibronectin fragments

Karen Pollok, Helmut Hanenberg, Timothy W. Noblitt, Wendy L. Schroeder, Ikunoshin Kato, David Emanuel, David A. Williams

Research output: Contribution to journalArticle

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Abstract

Primary human T lymphocytes are powerful targets for genetic modification, although the use of these targets in human gene therapy protocols has been hampered by low levels of transduction. We have shown previously that significant increases in the transduction of hematopoietic stem and progenitor cells with retroviral vectors can be obtained by the colocalization of the retrovirus and target cells on specific fibronectin (FN) adhesion domains (H. Hanenberg, X. L. Xiao, D. Dilloo, K. Hashino, I. Kato, and D. A. Williams, Nat. Med. 2:876-882, 1996). We studied the transfer of genes into primary T lymphocytes by using FN-assisted retroviral gene transfer. Activated T lymphocytes were infected for three consecutive days on the recombinant FN fragment CH-296 with a retroviral vector encoding the murine B7-1 protein. Transduced lymphocytes were analyzed for murine B7-1 expression, and it was found that under optimal conditions, 80 to 89% of the CD3+ lymphocytes were transduced. Gene transfer was predominantly augmented by the interaction between VLA-4 on the T lymphocytes and the FN adhesion site CS-1. Adenosine deaminase (ADA)-deficient primary T lymphocytes transduced on CH-296 with a retrovirus encoding murine ADA (mADA) exhibited levels of mADA activity several-fold higher than the levels of the endogenous human ADA protein observed in normal human T lymphocytes. Strikingly, the long-term expression of the transgene was dependent on the activation status of the lymphocytes. This approach will have important applications in human gene therapy protocols targeting primary T lymphocytes.

Original languageEnglish
Pages (from-to)4882-4892
Number of pages11
JournalJournal of Virology
Volume72
Issue number6
StatePublished - Jun 1998

Fingerprint

adenosine deaminase
Adenosine Deaminase
fibronectins
Fibronectins
gene transfer
T-lymphocytes
T-Lymphocytes
Retroviridae
Genes
retroviral vectors
gene therapy
lymphocytes
mice
Hematopoietic Stem Cells
Genetic Therapy
adhesion
Lymphocytes
Integrin alpha4beta1
Lymphocyte Activation
Transgenes

ASJC Scopus subject areas

  • Immunology

Cite this

High-efficiency gene transfer into normal and adenosine deaminase- deficient T lymphocytes is mediated by transduction on recombinant fibronectin fragments. / Pollok, Karen; Hanenberg, Helmut; Noblitt, Timothy W.; Schroeder, Wendy L.; Kato, Ikunoshin; Emanuel, David; Williams, David A.

In: Journal of Virology, Vol. 72, No. 6, 06.1998, p. 4882-4892.

Research output: Contribution to journalArticle

Pollok, Karen ; Hanenberg, Helmut ; Noblitt, Timothy W. ; Schroeder, Wendy L. ; Kato, Ikunoshin ; Emanuel, David ; Williams, David A. / High-efficiency gene transfer into normal and adenosine deaminase- deficient T lymphocytes is mediated by transduction on recombinant fibronectin fragments. In: Journal of Virology. 1998 ; Vol. 72, No. 6. pp. 4882-4892.
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