Generation of human parallel chimeric antigen receptor (pCAR) T cells to achieve synergistic T cell co-stimulation

• Published in: STAR protocols Vol. 3; no. 2; p. 101414
• Main Authors: Larcombe-Young, Daniel, Whilding, Lynsey, Davies, David Marc, Draper, Benjamin, Bechman, Natasha, Maher, John
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.06.2022; Elsevier
• Subjects: Cancer; Cell Biology; Cell isolation; Cell-based Assays; Flow Cytometry/Mass Cytometry; Humans; Immunology; Molecular Biology; Protocol; Receptors, Chimeric Antigen - genetics; Retroviridae; T-Lymphocytes; Immunology; Molecular Biology; Flow Cytometry/Mass Cytometry; Cell-based Assays; Cell isolation; Cell Biology; Cancer
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101414

Dual co-stimulation may be harnessed using parallel chimeric antigen receptors (pCARs) in which two distinct co-stimulatory units are adjacently localized on the plasma membrane. This protocol summarizes construct design, human T cell isolation, retroviral transduction, tissue culture expansion, and preclinical testing of pCAR T cells, exemplified by receptors that co-target avb6 integrin and ErbB dimers. For complete details on the use and execution of this protocol, please refer to Muliaditan et al. (2021). [Display omitted] •A protocol to enable robust generation of pCAR T cells•Effective dual co-stimulation is provided by this platform•Sustained and enhanced antitumor activity by pCAR T cells•Detailed in vitro and in vivo evaluation of pCAR technology Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Dual co-stimulation may be harnessed using parallel chimeric antigen receptors (pCARs) in which two distinct co-stimulatory units are adjacently localized on the plasma membrane. This protocol summarizes construct design, human T cell isolation, retroviral transduction, tissue culture expansion, and preclinical testing of pCAR T cells, exemplified by receptors that co-target avb6 integrin and ErbB dimers.