Retrovirus-mediated transfer of the herpes simplex virus thymidine kinase and enhanced green fluorescence protein genes in primary T lymphocytes.

Abstract:

:The EGFP-tk retroviral vector, encoding enhanced green fluorescent protein (EGFP) and the herpes simplex virus thymidine kinase (HSV-tk) packaged in a Phoenix amphotropic cell line, was used to transduce healthy donor T lymphocytes. Infection yielded a mean of 41.8 +/- 9.3% SD (range 31.1-48.4%) EGFP-positive cells and a mean of 92 +/- 2% SD (range 90-94%) after cell sorting. EGFP expression remained stable for 30 d after infection. The entire gene transfer procedure had no significant effect on lymphocyte subsets and slightly reduced clonogenicity. Ganciclovir (gcv) treatment (1 microg/ml x 10 d) killed all EGFP-positive cells in the transduced and transduced/sorted populations, but had no effect on untransduced controls. Our results show that primary T lymphocytes can be transduced using an EGFP-tk vector that yields a homogeneous infected population without affecting lymphocyte subsets, function and clonogenicity.

journal_name

Br J Haematol

authors

Di Florio S,Sebastiani C,Fagioli M,Di Ianni M,Alfonsi D,Venditti G,Pelicci PG,Tabilio A

doi

10.1046/j.1365-2141.2000.02264.x

subject

Has Abstract

pub_date

2000-09-01 00:00:00

pages

903-6

issue

4

eissn

0007-1048

issn

1365-2141

pii

bjh2264

journal_volume

110

pub_type

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