title: Generation of CD8(+) T cells expressing two additional T-cell receptors (TETARs) for personalised melanoma therapy.
abstract: Adoptive T-cell therapy of cancer often fails due to the tumor cells' immune escape mechanisms, like antigen loss or down-regulation. To anticipate immune escape by loss of a single antigen, it would be advantageous to equip T cells with multiple specificities. To study the possible interference of 2 T-cell receptors (TCRs) in one cell, and to examine how to counteract competing effects, we generated TETARs, CD8(+) T cells expressing two additional T-cell receptors by simultaneous transient transfection with 2 TCRs using RNA electroporation. The TETARs were equipped with one TCR specific for the common melanoma antigen gp100 and one TCR recognizing a patient-specific, individual mutation of CCT6A (chaperonin containing TCP1, subunit 6A) termed "CCT6A(m) TCR." These CD8(+) T cells proved functional in cytokine secretion and lytic activity upon stimulation with each of their cognate antigens, although some reciprocal inhibition was observed. Murinisation of the CCT6A(m) TCR increased and prolonged its expression and increased the lytic capacity of the dual-specific T cells. Taken together, we generated functional, dual-specific CD8(+) T cells directed against a common melanoma-antigen and an individually mutated antigen for the use in personalised adoptive T-cell therapy of melanoma. The intended therapy would involve repetitive injections of the RNA-transfected cells to overcome the transiency of TCR expression. In case of autoimmunity-related side effects, a cessation of treatment would result in a disappearance of the introduced receptors, which increases the safety of this approach.
authors.affiliation: a Department of Dermatology ; Universitätsklinikum Erlangen ; Erlangen , Germany.|b Department of Genetics ; Friedrich-Alexander-Universität Erlangen-Nürnberg ; Erlangen , Germany.|e SH and SP share first authorship.
authors.forename: Sandra
authors.initials: S
authors.lastname: Höfflin
authors.affiliation: a Department of Dermatology ; Universitätsklinikum Erlangen ; Erlangen , Germany.|b Department of Genetics ; Friedrich-Alexander-Universität Erlangen-Nürnberg ; Erlangen , Germany.|e SH and SP share first authorship.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Prommersberger
authors.affiliation: a Department of Dermatology ; Universitätsklinikum Erlangen ; Erlangen , Germany.
authors.forename: Ugur
authors.initials: U
authors.lastname: Uslu
authors.affiliation: a Department of Dermatology ; Universitätsklinikum Erlangen ; Erlangen , Germany.
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
authors.affiliation: c Cancer Immunotherapy Laboratory; QIMR Berghofer Medical Research Institute ; Brisbane , Queensland , Australia.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Schmidt
authors.affiliation: d Dept. Internal Medicine III ; Universitätsmedizin der Johannes Gutenberg-Universität Mainz ; Mainz , Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Lennerz
authors.affiliation: a Department of Dermatology ; Universitätsklinikum Erlangen ; Erlangen , Germany.|f JD and NS share senior authorship.
authors.forename: Jan
authors.initials: J
authors.lastname: Dörrie
authors.affiliation: a Department of Dermatology ; Universitätsklinikum Erlangen ; Erlangen , Germany.|f JD and NS share senior authorship.
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D045744:Cell Line, Tumor; D056404:Chaperonin Containing TCP-1 / Q000276:immunology; D006801:Humans; D016219:Immunotherapy, Adoptive; D008545:Melanoma / Q000276:immunology / Q000628:therapy*; D057285:Precision Medicine; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D012878:Skin Neoplasms / Q000276:immunology / Q000628:therapy*; D058951:gp100 Melanoma Antigen / Q000276:immunology
chemical_list: C533302:CCT6A protein, human; D011948:Receptors, Antigen, T-Cell; D058951:gp100 Melanoma Antigen; D056404:Chaperonin Containing TCP-1
keywords: Adoptive T-cell therapy; TCR transfer; anti-tumor activity; immune escape; immunotherapy; melanoma; neo-antigen