title: Nonviral RNA transfection to transiently modify T cells with chimeric antigen receptors for adoptive therapy.
abstract: Redirecting T cells with a chimeric antigen receptor (CAR) of predefined specificity showed remarkable efficacy in the adoptive therapy trials of malignant diseases. The CAR consists of a single chain fragment of variable region (scFv) antibody targeting domain covalently linked to the CD3ζ signalling domain of the T cell receptor complex to mediate T cell activation upon antigen engagement. By using an antibody-derived targeting domain a CAR can potentially redirect T cells towards any target expressed on the cell surface as long as a binding domain is available. Antibody-mediated targeting moreover circumvents MHC restriction of the targeted antigen, thereby broadening the potential of applicability of adoptive T cell therapy. While T cells were so far genetically modified by viral transduction, transient modification with a CAR by RNA transfection gained increasing interest during the last years. This chapter focuses on methods to modify human T cells from peripheral blood with a CAR by electroporation of in vitro transcribed RNA and to test modified T cells for function for use in adoptive immunotherapy.
authors.affiliation: Department I of Internal Medicine, Tumor Genetics, and Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Riet
authors.forename: Astrid
authors.initials: A
authors.lastname: Holzinger
authors.forename: Jan
authors.initials: J
authors.lastname: Dörrie
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
authors.forename: Hinrich
authors.initials: H
authors.lastname: Abken
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D017252:CD3 Complex / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D060846:Cell Engineering / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D008213:Lymphocyte Activation / Q000235:genetics / Q000276:immunology; D011946:Receptors, Antigen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D017252:CD3 Complex; C121704:CD3 antigen, zeta chain; D011946:Receptors, Antigen; D011993:Recombinant Fusion Proteins