Multifactorial optimization of gammaretroviral gene transfer into human T lymphocytes for clinical application Journal Article


Authors: Quintas-Cardama, A.; Yeh, R. K.; Hollyman, D.; Stefanski, J.; Taylor, C.; Nikhamin, Y.; Imperato, G.; Sadelain, M.; Riviere, I.; Brentjens, R. J.
Article Title: Multifactorial optimization of gammaretroviral gene transfer into human T lymphocytes for clinical application
Abstract: The ability to genetically modify human T cells to target tumor antigens through retroviral gene transfer constitutes a potentially powerful approach to cancer immunotherapy. However, low transduction efficiencies may hamper the efficacy of such therapeutic strategies in the clinical setting. Most commonly, gammaretroviral gene transfer into T cells is conducted through spinoculation, that is, centrifugation of retroviral particles and T cells on RetroNectin-coated non-tissue culture vessels. Here we present data investigating the impact of temperature, speed, and frequency of spinoculation on T cell transduction efficiencies. We found that all three variables independently impacted gene transfer, with increasing temperature, speed, and frequency of spinoculation all enhancing the transduction of T cells. These improved conditions were additive, with the greatest proportion of transduced T cells being generated at the highest tested temperature and speed, after daily spinoculation for 2 to 3 days. Under these conditions, enhanced gene transfer was observed in T cells derived from healthy donors, using research-grade vector stocks. Whereas both RetroNectin and spinoculation were critical to optimal gene transduction, preloading of gammaretroviral particles before spinoculation did not enhance gene transfer. Significantly, application of these enhanced transduction conditions to T cells derived from previously treated patients with chronic lymphocytic leukemia allowed for adequate gene transfer under both small-scale and large-scale clinically applicable conditions using either preclinical or current Good Manufacturing Practice-grade gammaretroviral vector stocks. © 2007 Mary Ann Liebert, Inc.
Keywords: controlled study; unclassified drug; human cell; t lymphocyte; t-lymphocytes; analytic method; viral gene delivery system; genetic transduction; genetic vectors; transduction, genetic; genetic transfection; gene therapy; temperature; chronic lymphatic leukemia; centrifugation; lymphocyte antigen receptor; retrovirus; retroviridae; virus vector; 192z1 chimeric antigen receptor
Journal Title: Human Gene Therapy
Volume: 18
Issue: 12
ISSN: 1043-0342
Publisher: Mary Ann Liebert, Inc  
Date Published: 2007-12-01
Start Page: 1253
End Page: 1260
Language: English
DOI: 10.1089/hum.2007.088
PUBMED: 18052719
PROVIDER: scopus
DOI/URL:
Notes: --- - "Cited By (since 1996): 10" - "Export Date: 17 November 2011" - "CODEN: HGTHE" - "Source: Scopus"
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