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Redirecting human T lymphocytes toward renal cell carcinoma specificity by retroviral transfer of T cell receptor genes

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Item Type:Article
Title:Redirecting human T lymphocytes toward renal cell carcinoma specificity by retroviral transfer of T cell receptor genes
Creators Name:Engels, B. and Noessner, E. and Frankenberger, B. and Blankenstein, T. and Schendel, D.J. and Uckert, W.
Abstract:Adoptive T cell therapy of renal cell carcinoma (RCC) is limited by the difficulty in generating sufficient numbers of RCC-reactive T cells in vitro. To circumvent this problem, we cloned T cell receptor (TCR) α and β chains from a tumor-infiltrating lymphocyte clone specific for an RCC tumor antigen and transferred the TCR into human T cell lines and primary T lymphocytes. Efficient TCR expression in primary T lymphocytes was obtained only with a mouse myeloproliferative sarcoma virus (MPSV)-based retroviral vector, not with a Moloney murine leukemia virus (MLV)-based vector, although both viral supernatants were similar in titer, as shown by analysis of copy number integration in transduced T cells. Reverse transcription-polymerase chain reaction analysis revealed a higher amount of TCR-encoding transcripts when T cells were transduced with the MPSV vector in comparison with the MLV vector, indicating that high TCR expression levels can be achieved by appropriate cis-regulatory vector elements. The biological activity of the transferred TCR was shown by specific lysis of RCC cells (51Cr release assay) and by interferon γ and tumor necrosis factor α release (enzyme-linked immunosorbent assay) in an antigen-specific and HLA-A*0201-restricted fashion. Comparison of the redirected T lymphocytes with the original tumor-infiltrating lymphocyte clone revealed similar killing and cytokine secretion capabilities. The functional activity of TCR-redirected T lymphocytes was stable over time. The results demonstrate that use of an optimized retroviral vector yielded a high TCR transduction efficiency and stable and high TCR expression in primary human T lymphocytes and redirected their specificity toward RCC cells.
Keywords:Renal Cell Carcinoma, Cell Line, Cultured Cells, Cytotoxic T-Lymphocytes, Gene Expression, Genetic Recombination, Genetic Transduction, Genetic Vectors, Interferon Type II, Kidney Neoplasms, Moloney Murine Leukemia Virus, Murine Sarcoma Viruses, Retroviridae, T-Cell Receptor Genes, Tumor Necrosis Factor-Alpha
Source:Human Gene Therapy
ISSN:1043-0342
Publisher:Mary Ann Liebert (U.S.A.)
Volume:16
Number:7
Page Range:799-810
Date:1 July 2005
Official Publication:https://doi.org/10.1089/hum.2005.16.799
PubMed:View item in PubMed

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