A GMP-compliant protocol to expand and transfect cancer patient T cells with mRNA encoding a tumor-specific chimeric antigen receptor

Abstract

Chimeric antigen receptors (CARs), which combine an antibody-derived binding domain (single chain fragment variable) with T-cell-activating signaling domains, have become a promising tool in the adoptive cellular therapy of cancer. Retro- and lenti-viral transductions are currently the standard methods to equip T cells with a CAR; permanent CAR expression, however, harbors several risks like uncontrolled auto-reactivity. Modification of T cells by electroporation with CAR-encoding RNA to achieve transient expression likely circumvents these difficulties. We here present a GMP-compliant protocol to activate and expand T cells for clinical application. The protocol is optimized in particular to produce CAR-modified T cells in clinically sufficient numbers under full GMP-compliance from late-stage cancer patients. This protocol allows the generation of 6.7 × 108 CAR-expressing T cells from one patient leukapheresis. The CAR-engineered T cells produced pro-inflammatory cytokines after stimulation with antigen-bearing tumor cells and lysed tumor cells in an antigen-specific manner. This functional capacity was maintained after cryopreservation. Taken together, we provide a clinically applicable protocol to transiently engineer sufficient numbers of antigen-specific patient T cells for use in adoptive cell therapy of cancer.

DOI: 10.1007/s00262-014-1572-5

2 Figures and Tables

0204060802014201520162017
Citations per Year

64 Citations

Semantic Scholar estimates that this publication has 64 citations based on the available data.

See our FAQ for additional information.

Cite this paper

@article{Krug2014AGP, title={A GMP-compliant protocol to expand and transfect cancer patient T cells with mRNA encoding a tumor-specific chimeric antigen receptor}, author={Christian Krug and Manuel Wiesinger and Hinrich Abken and Beatrice Schuler-Thurner and Gerold Schuler and Jan D{\"{o}rrie and Niels Schaft}, journal={Cancer Immunology, Immunotherapy}, year={2014}, volume={63}, pages={999-1008} }