Comparison of lentiviral and sleeping beauty mediated αβ T cell receptor gene transfer

• Published in: PloS one Vol. 8; no. 6; p. e68201
• Main Authors: Field, Anne-Christine, Vink, Conrad, Gabriel, Richard, Al-Subki, Roua, Schmidt, Manfred, Goulden, Nicholas, Stauss, Hans, Thrasher, Adrian, Morris, Emma, Qasim, Waseem
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 28.06.2013; Public Library of Science (PLoS)
• Subjects: Accelerated tests; Animals; Antigens; Biology; Biotechnology; Cancer; Cell Line, Tumor; Childrens health; Chimeric antigen receptors; College campuses; Cytokines; DNA Transposable Elements - genetics; Ethics; Exploitation; Flow cytometry; Gene expression; Gene therapy; Gene transfer; Gene Transfer Techniques; Genes, T-Cell Receptor - genetics; Genetic Therapy - methods; Genetic Vectors - genetics; HIV; Human immunodeficiency virus; Humans; Immunoglobulins; Immunology; In vivo methods and tests; Integration; Jurkat Cells; Lentivirus - genetics; Lymphocytes; Lymphocytes T; Lysis; Medical research; Medicine; Mice; Mice, Inbred C57BL; Penicillin; Peptides; Plasmids - genetics; Receptors; Receptors, Antigen, T-Cell - genetics; Recombinant Fusion Proteins - genetics; Rodents; System effectiveness; T cell receptors; T-cell receptor; T-Lymphocytes - metabolism; Transcription; Transfection; Transgenes; Transposase; Transposition; Transposons; Tumors; Vectors (Biology); United Kingdom > UK
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0068201

Transfer of tumour antigen-specific receptors to T cells requires efficient delivery and integration of transgenes, and currently most clinical studies are using gamma retroviral or lentiviral systems. Whilst important proof-of-principle data has been generated for both chimeric antigen receptors and αβ T cell receptors, the current platforms are costly, time-consuming and relatively inflexible. Alternative, more cost-effective, Sleeping Beauty transposon-based plasmid systems could offer a pathway to accelerated clinical testing of a more diverse repertoire of recombinant high affinity T cell receptors. Nucleofection of hyperactive SB100X transposase-mediated stable transposition of an optimised murine-human chimeric T cell receptor specific for Wilm's tumour antigen from a Sleeping Beauty transposon plasmid. Whilst transfer efficiency was lower than that mediated by lentiviral transduction, cells could be readily enriched and expanded, and mediated effective target cells lysis in vitro and in vivo. Integration sites of transposed TCR genes in primary T cells were almost randomly distributed, contrasting the predilection of lentiviral vectors for transcriptionally active sites. The results support exploitation of the Sleeping Beauty plasmid based system as a flexible and adaptable platform for accelerated, early-phase assessment of T cell receptor gene therapies.