Manufacture of T cells using the Sleeping Beauty system to enforce expression of a CD19-specific chimeric antigen receptor

• Published in: Cancer gene therapy Vol. 22; no. 2; pp. 95 - 100
• Main Authors: Singh, H, Moyes, J S E, Huls, M H, Cooper, L J N
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01-02-2015; Nature Publishing Group
• Subjects: 13; 13/100; 631/67/71; 631/80; Antigen (tumor-associated); Antigen-Presenting Cells; Antigens; Antigens, CD19 - genetics; Antigens, CD19 - immunology; Antigens, CD19 - therapeutic use; Biomedical and Life Sciences; Biomedicine; CD19 antigen; CD3 antigen; Cell proliferation; Cell receptors; Cell surface; Chimeric antigen receptors; Gene Expression; Gene Therapy; Gene transfer; Gene Transfer Techniques; Genetic aspects; Genetic research; Histocompatibility antigen HLA; Humans; Immunotherapy, Adoptive; Lymphocyte Activation - immunology; Lymphocytes; Lymphocytes T; Properties; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - immunology; Receptors, Antigen, T-Cell - therapeutic use; Retroelements - genetics; review; T cell receptors; T cells; T-Lymphocytes - immunology
• ISSN: 0929-1903; 1476-5500
• DOI: 10.1038/cgt.2014.69

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.