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Redirecting Human T Lymphocytes Toward Renal Cell Carcinoma Specificity by Retroviral Transfer of T Cell Receptor Genes

2005; Mary Ann Liebert, Inc.; Volume: 16; Issue: 7 Linguagem: Inglês

10.1089/hum.2005.16.799

1557-7422

Boris Engels, Elfriede Noeßner, Bernhard Frankenberger, Thomas Blankenstein, Dolores J. Schendel, Wolfgang Uckert,

Immunotherapy and Immune Responses

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) ␣ and ␤ 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 ( 51 Cr release assay) and by interferon ␥ and tumor necrosis factor ␣ 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.799 OVERVIEW SUMMARY The therapeutic efficacy of adoptively transferred cytotoxic T lymphocytes (CTLs) has been demonstrated in clinical trials.It is, however, limited by the difficulty in generating sufficient amounts of CTLs in vitro.Gene therapy approaches include the genetic engineering of T cell specificity by T cell receptor (TCR) gene transfer.It was demonstrated that the number of TCR molecules on the T cell is important for its function.An efficient transfer system that yields high transduction efficiency and strong and stable transgene expression is a prerequisite to achieve effector function for redirected T cells.We used an improved retroviral vector and transferred a renal cell carcinoma (RCC)-reactive TCR.The redirected peripheral blood lymphocytes of healthy donors gained specificity for an RCC tumor antigen.The reprogrammed T cells showed cytokine secretion and tumor cell killing similar to the original tumor-infiltrating lymphocyte clone.the few immunogenic tumor entities.In some patients, par-