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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

Paper ID Volume ID Publish Year Pages File Format Full-Text
23740 43468 2011 9 PDF Available
Title
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
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 (0.5 μM) 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 1 μM, 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.

► Proteasome inhibition enhances LV transduction of HSC and T cells. ► Proteasome inhibition facilitates LV transduction with large size vectors. ► Proteasome inhibition does not affect viability or functions of transduced cells.

Keywords
CB, cord blood; eGFP, enhanced green fluorescent protein; FL, flt3/flk2 ligand; HSC, hematopoietic stem cells; HLA, human leukocyte antigen; IL-6, interleukin-6; IL-2, interleukin-2; LV, lentiviral vectors; MLR, mixed lymphocyte reaction; MOI, multiplicit
First Page Preview
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
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 156, Issue 3, 10 December 2011, Pages 218–226
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering