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
| Autor: | Dario Sangiolo, Alessandro Cignetti, Giulia Mesiano, Loretta Gammaitoni, Noela Jordaney, Cristina Cammarata, Valeria Leuci, Wanda Piacibello, Massimo Aglietta, Sonia Capellero, Angela Rita Elia, Paola Circosta, Maja Todorovic, George E. Georges, Franca Fagioli, Marina Lesnikova |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
T-Lymphocytes
Genetic Vectors Antigens CD34 Bioengineering Lentiviral transduction Human leukocyte antigen Biology Real-Time Polymerase Chain Reaction Applied Microbiology and Biotechnology Peripheral blood mononuclear cell Transduction efficiency Hematopoietic stem cells chemistry.chemical_compound Transduction (genetics) Transduction Genetic MG132 medicine Humans Cells Cultured Lentivirus General Medicine Flow Cytometry Molecular biology Haematopoiesis chemistry Proteasome Proteasome inhibitor Stem cell Proteasome Inhibitors Biotechnology medicine.drug |
| Zdroj: | Journal of Biotechnology. 156:218-226 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/j.jbiotec.2011.09.001 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect