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Retroviral vectors for high-level transgene expression in T lymphocytes

Item Type:Article
Title:Retroviral vectors for high-level transgene expression in T lymphocytes
Creators Name:Engels, B. and Cam, H. and Schueler, T. and Indraccolo, S. and Gladow, M. and Baum, C. and Blankenstein, T. and Uckert, W.
Abstract:Efficient expression of genes transferred by retroviral vectors is a prerequisite for gene therapy, especially when the biological effect depends on the amount of transgene product. High-level gene expression is desirable for several gene therapy approaches involving T lymphocytes. We evaluated standard retroviral vectors with cis-regulatory control elements of the Moloney murine leukemia virus (Mo-MLV) with or without the human T cell-specific CD2 enhancer. For comparison, vectors containing the long terminal repeat (LTR) of myeloproliferative sarcoma virus (MPSV) and an improved 5′ untranslated region were used (MP71 vectors), with or without the woodchuck hepatitis virus posttranscriptional regulatory element (PRE). All vectors expressed the enhanced green fluorescent protein (GFP) to measure transgene expression. In mouse T cells MP71 vectors with and without the PRE yielded an up to 10-fold higher expression level compared with the MoMLV-based vectors currently used for gene transfer into T lymphocytes. A high multiplicity of infection (MOI) of standard Mo-MLV vectors could not reach expression levels obtained with a low MOI of MP71 vector. Ex vivo-transduced mouse T lymphocytes maintained the vector-dependent differences in level of transgene expression in Rag-1-deficient mice when adoptively transferred. In four human T cell lines and human primary T lymphocytes MP71 vectors yielded an up to 75-fold higher GFP expression level in comparison with the standard Mo-MLV vector. In contrast to mouse T cells, the integration of the PRE into MP71 vectors induced in human T cells a further significant increase in transgene expression level. Southern blot analysis of CEM T cells revealed that the superior performance of MP71 vectors was not due to a higher rate of viral integration. In summary, MP71 vectors are useful tools for stable, high-level gene expression in T lymphocytes, for example, in the expression of T cell receptor genes.
Keywords:3T3 Cells, CD2 Antigens, Cell Line, Cell Survival, Cultured Cells, Enhancer Elements, Gene Expression, Gene Therapy, Genetic Transduction, Genetic Vectors, Green Fluorescent Proteins, Luminescent Proteins, Moloney Murine Leukemia Virus, Nucleic Acid Regulatory Sequences, Retroviridae, Reverse Transcriptase Polymerase Chain Reaction, T-Lymphocytes, Transgenes, Animals, Mice
Source:Human Gene Therapy
Publisher:Mary Ann Liebert
Page Range:1155-1168
Date:10 August 2003
Official Publication:https://doi.org/10.1089/104303403322167993
PubMed:View item in PubMed

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