Transduction of primary lymphocytes with Epstein-Barr virus (EBV) latent membrane protein-specific T-cell receptor induces lysis of virus-infected cells: A novel strategy for the treatment of Hodgkin's disease and nasopharyngeal carcinoma.
Adoptive immunotherapy with in vitro expanded cytotoxic T lymphocytes specific for Epstein-Barr virus (EBV) can successfully treat post-transplant lymphoproliferative disease (PTLD). However, extension of a similar strategy to Hodgkin's disease (HD) and nasopharyngeal carcinoma (NPC) is limited by the poor immunogenicity of the limited set of EBV latency antigens expressed in these malignancies, making T-cell expansion difficult. Retroviral transduction of LMP-specific T-cell receptors (TCR) into activated T lymphocytes may provide a universal, MHC-restricted, means to generate effector cells without the need for tissue culture based methods of CTL expansion. We report the transfer of two LMP2-specific TCRs from human T-cell clones (HLA-A2 and HLA-A23,24 restricted) that confer the ability to lyse EBV-immortalized B-lymphoblastoid cell lines (B-LCL). B-LCL are the best model for native expression of LMP2. We also demonstrate the rapid transfer of the TCR by nucleofection of primary T cells using a simple plasmid-based vector. The ability to detect nucleofected TCRVbeta chain by antibody, fully assembled TCR by tetramer, and peptide-MHC-specific lytic activity indicates that nucleofection can serve as a tool for rapid screening of TCR specificity.
Jurgens
Lisa A
LA
Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
Khanna
Rajiv
R
Weber
James
J
Orentas
Rimas J
RJ
D000595:Amino Acid Sequence; D017951:Antigen Presentation; D019002:CD56 Antigen / Q000378:metabolism; D045744:Cell Line, Tumor; D015234:HLA-A Antigens; D006689:Hodgkin Disease / Q000628:therapy*; D006801:Humans; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000821:virology*; D008969:Molecular Sequence Data; D009303:Nasopharyngeal Neoplasms / Q000628:therapy*; D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014161:Transduction, Genetic; D014763:Viral Matrix Proteins / Q000378:metabolism* / Q000502:physiology
D019002:CD56 Antigen; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D015234:HLA-A Antigens; D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta; D014763:Viral Matrix Proteins