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Highly‐efficient gene transfer with retroviral vectors into human T lymphocytes on fibronectin
Author(s) -
Fehse Boris,
Schade Ulrika M,
LI Zhixiong,
Uhde Almut,
Koch Stefan,
Goller Bernhard,
RÜger RÜdiger,
Fehse Natalia,
StochschlÄder Marcus,
Zander Axel R
Publication year - 1998
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1998.00785.x
Subject(s) - transduction (biophysics) , biology , jurkat cells , viral vector , thymidine kinase , virology , genetic enhancement , microbiology and biotechnology , ganciclovir , genetically modified organism , transfection , cell sorting , gene , herpes simplex virus , immunology , virus , t cell , human cytomegalovirus , recombinant dna , genetics , flow cytometry , immune system , biochemistry
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.