Universität zu Köln

Krug, Christian, Wiesinger, Manuel, Abken, Hinrich, Schuler-Thurner, Beatrice, Schuler, Gerold, Dorrie, Jan ORCID: 0000-0002-3478-0741 and Schaft, Niels ORCID: 0000-0001-8236-9298 (2014). A GMP-compliant protocol to expand and transfect cancer patient T cells with mRNA encoding a tumor-specific chimeric antigen receptor. Cancer Immunol. Immunother., 63 (10). S. 999 - 1009. NEW YORK: SPRINGER. ISSN 1432-0851

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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 x 10(8) 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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Krug, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Wiesinger, Manuel UNSPECIFIED UNSPECIFIED UNSPECIFIED Abken, Hinrich UNSPECIFIED UNSPECIFIED UNSPECIFIED Schuler-Thurner, Beatrice UNSPECIFIED UNSPECIFIED UNSPECIFIED Schuler, Gerold UNSPECIFIED UNSPECIFIED UNSPECIFIED Dorrie, Jan UNSPECIFIED orcid.org/0000-0002-3478-0741 UNSPECIFIED Schaft, Niels UNSPECIFIED orcid.org/0000-0001-8236-9298 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-427586
DOI:	10.1007/s00262-014-1572-5
Journal or Publication Title:	Cancer Immunol. Immunother.
Volume:	63
Number:	10
Page Range:	S. 999 - 1009
Date:	2014
Publisher:	SPRINGER
Place of Publication:	NEW YORK
ISSN:	1432-0851
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ADOPTIVE IMMUNOTHERAPY; RECOMBINANT IMMUNORECEPTORS; ACTIVATION; ELUTRIATION; MELANOMA; THERAPY Multiple languages Oncology; Immunology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/42758