title: Generation of diffuse large B cell lymphoma-associated antigen-specific Vα6/Vβ13+T cells by TCR gene transfer.
abstract: BACKGROUND
  Our previous study had amplified antigen-specific full-length TCR α and β 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.
  
  
  METHODS
  Two different eukaryotic expression plasmids harboring TCR Vα6 and TCR Vβ13 genes specific for DLBCL-associated antigens were constructed and subsequently transferred into human T cells using Nucleofector™ technique. The expression of targeted genes in TCR gene-modified cells was detected by real-time PCR, and western blot using TCR Vβ 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 Vα6 and TCR Vβ13 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 Vβ13-IRES-TCR Vα6 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.
authors.affiliation: Institute of Hematology, Medical College, Jinan University, Guangzhou, 510632, PR China.
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Yin
authors.forename: Xianfeng
authors.initials: X
authors.lastname: Zha
authors.forename: Lijian
authors.initials: L
authors.lastname: Yang
authors.forename: Shaohua
authors.initials: S
authors.lastname: Chen
authors.forename: Yubing
authors.initials: Y
authors.lastname: Zhou
authors.forename: Xiuli
authors.initials: X
authors.lastname: Wu
authors.forename: Yangqiu
authors.initials: Y
authors.lastname: Li
mesh_terms: D045744:Cell Line, Tumor; D018014:Gene Transfer Techniques; D006801:Humans; D016393:Lymphoma, B-Cell / Q000235:genetics* / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000276:immunology; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D016693:Receptors, Antigen, T-Cell, alpha-beta