Transduction efficiency of MLV but not of HIV-1 vectors is pseudotype dependent on human primary T lymphocytes

Autor: Wolfgang Uckert, Klaus Cichutek, Matthias Schweizer, Stefanie Steidl, Jörn Stitz, Michael D. Mühlebach, Isabel Schmitt, Thomas Blankenstein
Rok vydání: 2003
Předmět:
Zdroj: Journal of Molecular Medicine. 81:801-810
ISSN: 1432-1440
0946-2716
Popis: The success of several gene therapeutic approaches requires efficient transduction of human primary T lymphocytes. For this it is important to enhance the transduction efficiency, and this can be achieved by various means, mainly technical development of transduction procedures and use of different vectors and vector pseudotypes. We analyzed the transduction efficiency of an HIV-1 vector encoding enhanced green fluorescent protein (GFP) as a marker gene and pseudotyped with the envelopes of MLV-A, MLV-10A1, GaLV, RD114, and VSV for human primary T lymphocytes in comparison to an MLV vector pseudotyped with the same envelopes. Pseudotyping of the MLV vector with the envelopes of 10A1 and GaLV resulted in efficient transduction of preactivated human primary T lymphocytes (32.4% and 32.7% CD3+/GFP+ cells, respectively) while MLV-A (14.0%), RD114 (8.8%), and VSV (1.5%) envelopes were less efficient when using titrated vector stocks equilibrated to a multiplicity of infection of 1. In contrast, the HIV-1 vectors pseudotyped with these envelope proteins transduced preactivated T lymphocytes with similar efficiency (approx. 20% CD3+/GFP+ cells). Thereby, CD4+ and CD8+ T lymphocyte subpopulations were transduced at equivalent levels. The similar performance of the different HIV-1 vector pseudotypes may be due in part to the similar half-lives of the vector particles. Independently of the envelope used for pseudotyping neither the MLV nor the HIV-1 vectors yielded any significant transduction in nonactivated T lymphocytes (below 0.55% of GFP+ cells)
Databáze: OpenAIRE
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