title: Comparison of lentiviral and sleeping beauty mediated αβ T cell receptor gene transfer.
abstract: 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.
authors.affiliation: Molecular immunology Unit, Institute of Child Health, University College London, London, United Kingdom.
authors.forename: Anne-Christine
authors.initials: AC
authors.lastname: Field
authors.forename: Conrad
authors.initials: C
authors.lastname: Vink
authors.forename: Richard
authors.initials: R
authors.lastname: Gabriel
authors.forename: Roua
authors.initials: R
authors.lastname: Al-Subki
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.forename: Nicholas
authors.initials: N
authors.lastname: Goulden
authors.forename: Hans
authors.initials: H
authors.lastname: Stauss
authors.forename: Adrian
authors.initials: A
authors.lastname: Thrasher
authors.forename: Emma
authors.initials: E
authors.lastname: Morris
authors.forename: Waseem
authors.initials: W
authors.lastname: Qasim
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004251:DNA Transposable Elements / Q000235:genetics*; D018014:Gene Transfer Techniques; D019672:Genes, T-Cell Receptor / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D019169:Jurkat Cells; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins