title: Manufacture of T cells using the Sleeping Beauty system to enforce expression of a CD19-specific chimeric antigen receptor.
abstract: T cells can be reprogrammed to redirect specificity to tumor-associated antigens (TAAs) through the enforced expression of chimeric antigen receptors (CARs). The prototypical CAR is a single-chain molecule that docks with TAA expressed on the cell surface and, in contrast to the T-cell receptor complex, recognizes target cells independent of human leukocyte antigen. The bioprocessing to generate CAR(+) T cells has been reduced to clinical practice based on two common steps that are accomplished in compliance with current good manufacturing practice. These are (1) gene transfer to stably integrate the CAR using viral and nonviral approaches and (2) activating the T cells for proliferation by crosslinking CD3 or antigen-driven numeric expansion using activating and propagating cells (AaPCs). Here, we outline our approach to nonviral gene transfer using the Sleeping Beauty system and the selective propagation of CD19-specific CAR(+) T cells on AaPCs.
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: H
authors.initials: H
authors.lastname: Singh
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: J S E
authors.initials: JS
authors.lastname: Moyes
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: M H
authors.initials: MH
authors.lastname: Huls
authors.affiliation: 1] Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA [2] The University of Texas Graduate School of Biomedical Sciences at UT MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: L J N
authors.initials: LJ
authors.lastname: Cooper
mesh_terms: D000938:Antigen-Presenting Cells; D018941:Antigens, CD19 / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D018014:Gene Transfer Techniques; D006801:Humans; D016219:Immunotherapy, Adoptive; D008213:Lymphocyte Activation / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D018626:Retroelements / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D018941:Antigens, CD19; D011948:Receptors, Antigen, T-Cell; D018626:Retroelements