Transient proteasome inhibition as a strategy to enhance lentiviral transduction of hematopoietic CD34 + cells and T lymphocytes: Implications for the use of low viral doses and large-size vectors

Valeria Leuci, Giulia Mesiano, Loretta Gammaitoni, Cristina Cammarata, Sonia Capellero, Maja Todorovic, Noela Jordaney, Paola Circosta, Angela Elia, Marina Lesnikova, George E. Georges, Wanda Piacibello, Franca Fagioli, Alessandro Cignetti, Massimo Aglietta, Dario Sangiolo*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

The proteasome system restricts lentiviral transduction of stem cells. We exploited proteasome inhibition as a strategy to enhance transduction of both hematopoietic stem cells (HSC) and T lymphocytes with low dose or large-size lentiviral vectors (LV). HSC showed higher transduction efficiency if transiently exposed to proteasome inhibitor MG132 (41.8% vs 10.7%, p<0.0001). Treatment with MG132 (0.5μM) retained its beneficial effect with 3 different LV of increasing size up to 10.9. Kb (p<0.01). We extended, for the first time, the application of proteasome inhibition to the transduction of T lymphocytes. A transient exposure to MG132 significantly improved lentiviral T-cell transduction. The mean percentage of transduced T cells progressively increased from 13.5% of untreated cells, to 21% (p=0.3), 30% (p=0.03) and 37% (p=0.01) of T lymphocytes that were pre-treated with MG132 at 0.1, 0.5 and 1μM, respectively. MG132 did not affect viability or functionality of HSC or T cells, nor significantly increased the number of integrated vector copies. Transient proteasome inhibition appears as a new procedure to safely enhance lentiviral transduction of HSC and T lymphocytes with low viral doses. This approach could be useful in settings where the use of large size vectors may impair optimal viral production.

Original languageEnglish (US)
Pages (from-to)218-226
Number of pages9
JournalJournal of Biotechnology
Volume156
Issue number3
DOIs
StatePublished - Dec 10 2011

Keywords

  • Hematopoietic stem cells
  • Lentiviral transduction
  • T lymphocytes
  • Transduction efficiency

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Bioengineering
  • Biotechnology

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