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

Sabrina Di Florio, Carla Sebastiani, Marta Fagioli, Mauro Di Ianni, Diego Alfonsi, Gigliola Venditti, Pier Giuseppe Pelicci, Antonio Tabilio

Research output: Contribution to journalArticle

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 μg/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.

Original languageEnglish
Pages (from-to)903-906
Number of pages4
JournalBritish Journal of Haematology
Volume110
Issue number4
DOIs
Publication statusPublished - 2000

Keywords

  • Gene transfer
  • Green fluorescent protein
  • T lymphocytes
  • Thymidine kinase

ASJC Scopus subject areas

  • Hematology

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