T-lymphocyte function after retroviral-mediated thymidine kinase gene transfer and G418 selection

Mauro Di Ianni, Sabrina Di Florio, Gigliola Venditti, Concetta Liberatore, Francesca Lucheroni, Franca Falzetti, Adelmo Terenzi, Carmelo Carlo Stella, Fabrizio Spinozzi, Patrice Mannoni, Massimo F. Martelli, Antonio Tabilio

Research output: Contribution to journalArticlepeer-review

Abstract

Generation of an efficient graft-versus-leukemia (GVL) effect in patients with hematological malignancies who relapse after allogeneic bone marrow transplantation depends in part upon the number of infused T lymphocytes. Currently, a GVL reaction cannot be achieved without inducing concomitant graft-versus-host disease (GVHD); thus, one strategy is to try to modulate this GVL/GVHD ratio. We engineered human T lymphocytes with herpes simplex virus-thymidine kinase and neomycin resistance genes, with an LXSN- derived vector that confers a ganciclovir-specific sensitivity to the transduced T cells. We analyzed proliferation, interleukin-2 production, alloreactivity in a mixed lymphocyte culture, and clonogenicity during the different stages of retroviral infection and G418 selection. Our results confirm that a sufficient number of transduced T lymphocytes can be obtained after selection for clinical studies. Their proliferative activity, alloresponsiveness, and ability to produce and respond to interleukin-2 were retained. Compared with control populations, their clonogenicity, as assessed by limiting dilution assays, was reduced after retroviral infection and G418 selection by 1.6 and 2.9 logs, respectively, with both viral supernatant incubation and coculture procedures. This study shows that infection and selection with the thymidine kinase-neomycin resistance gene retroviral vector significantly reduces the number of functional T lymphocytes. This finding should be taken into account when establishing the dose of T lymphocytes necessary to trigger a modulated GVL/GVHD effect.

Original languageEnglish
Pages (from-to)920-926
Number of pages7
JournalCancer Gene Therapy
Volume7
Issue number6
Publication statusPublished - 2000

Keywords

  • Gene transfer
  • Limiting dilution assay
  • T lymphocytes
  • Thymidine kinase

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

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