Journal
JOURNAL OF HEMATOLOGY & ONCOLOGY
Volume 4, Issue -, Pages -Publisher
BMC
DOI: 10.1186/1756-8722-4-2
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Funding
- National 863 projects of China [2006AA02Z114]
- National Natural Science Foundation of Guangdong Province [05103293, 9251063201000001]
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Background: Our previous study had amplified antigen-specific full-length TCR alpha and beta genes of clonally expanded T cells in the peripheral blood (PB) of patients with diffuse large B-cell lymphoma (DLBCL). The transfer of T cell receptor (TCR) genes endows T cells with new antigen specificity. Therefore, the aim of this study is to generate diffuse large B cell lymphoma (DLBCL)-specific T cells by T cell receptor (TCR) gene transfer. Materials and methods: Two different eukaryotic expression plasmids harboring TCR V alpha 6 and TCR V beta 13 genes specific for DLBCL-associated antigens were constructed and subsequently transferred into human T cells using Nucleofector (TM) technique. The expression of targeted genes in TCR gene-modified cells was detected by real-time PCR, and western blot using TCR V beta antibody. The specific cytotoxicity of TCR gene-transferred T cells in vitro was estimated using a lactate dehydrogenase (LDH) release assay. Results: Two different eukaryotic expression plasmids harboring TCR V alpha 6 and TCR V beta 13 genes specific for DLBCL-associated antigens were constructed and subsequently transferred into T cells from healthy donors. Specific anti-DLBCL cytotoxic T lymphocytes (CTL) could be induced by transduction of specific TCR gene to modify healthy T cells. The transgene cassette of TCR V beta 13-IRES-TCR V alpha 6 was superior to the other in the function of TCR-redirected T cells. Conclusions: Specific anti-DLBCL cytotoxic T lymphocyte (CTL) could be inducted by transduction of specific TCR gene to modify healthy T cells.
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