Highly‐efficient gene transfer with retroviral vectors into human T lymphocytes on fibronectin

Genetically modified lymphocytes have been successfully used for correction of ADA deficiency in children and in controlling graft‐versus‐host disease (GvHD) after allogeneic bone marrow transplantation. Low transduction efficiencies are, however, limiting for gene therapeutic strategies based on ly...

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Bibliographic Details
Published in:British journal of haematology 1998-07, Vol.102 (2), p.566-574
Main Authors: Fehse, Boris, Schade, Ulrika M, LI, Zhixiong, Uhde, Almut, Koch, Stefan, Goller, Bernhard, RÜger, RÜdiger, Fehse, Natalia, StochschlÄder, Marcus, Zander, Axel R
Format: Article
Language:English
Subjects:
Anti-Allergic Agents - metabolism
Biological and medical sciences
Biotechnology
fibronectin
Fibronectins - metabolism
Fundamental and applied biological sciences. Psychology
Ganciclovir - pharmacology
Gene therapy
Gene Transfer Techniques
Genetic Vectors - genetics
Health. Pharmaceutical industry
Hematology
Humans
Industrial applications and implications. Economical aspects
Integrin alpha4beta1
Integrins - metabolism
Jurkat Cells
Receptors, Fibronectin - metabolism
Receptors, Lymphocyte Homing - metabolism
Receptors, Nerve Growth Factor - metabolism
retroviral vectors
Retroviridae - genetics
T lymphocytes
T-Lymphocytes - virology
Thymidine Kinase - genetics
ΔLNGFR
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Summary:Genetically modified lymphocytes have been successfully used for correction of ADA deficiency in children and in controlling graft‐versus‐host disease (GvHD) after allogeneic bone marrow transplantation. Low transduction efficiencies are, however, limiting for gene therapeutic strategies based on lymphocytes. In this study we compared protocols for highly efficient gene transfer into human T cells using retroviral vector‐containing supernatant. We showed that infection of both human primary T cells and CD4+ Jurkat cells is most efficient on the matrix component fibronectin. Transduction was carried out with a retroviral vector encoding both the human intracytoplasmatically truncated low‐affinity nerve growth factor receptor (ΔLNGFR) as a gene transfer marker and the Herpes simplex virus thymidine kinase for negative selection. Based on ΔLNGFR expression genetically modified cells were enriched to near purity by magnetic cell sorting (MACS). Enriched cells could be shown to be highly sensitive to ganciclovir.
ISSN:0007-1048
1365-2141
DOI:10.1046/j.1365-2141.1998.00785.x