Knies, Diana and Klobuch, Sebastian and Xue, Shao-An and Birtel, Matthias and Echchannaoui, Hakim and Yildiz, Oezlem and Omokoko, Tana and Guillaume, Philippe and Romero, Pedro and Stauss, Hans and Sahin, Ugur and Herr, Wolfgang and Theobald, Matthias and Thomas, Simone and Voss, Ralf-Holger (2016) An optimized single chain TCR scaffold relying on the assembly with the native CD3-complex prevents residual mispairing with endogenous TCRs in human T-cells. ONCOTARGET, 7 (16). pp. 21199-21221. ISSN 1949-2553,
Full text not available from this repository. (Request a copy)Abstract
Immunotherapy of cancer envisions the adoptive transfer of T-cells genetically engineered with tumor-specific heterodimeric alpha/beta T-cell receptors (TCR alpha/beta). However, potential mispairing of introduced TCR alpha/beta-chains with endogenous beta/alpha-ones may evoke unpredictable autoimmune reactivities. A novel single chain (sc) TCR format relies on the fusion of the V alpha-Linker-V beta-fragment to the TCR C beta-domain and coexpression of the TCR C alpha-domain capable of recruiting the natural CD3-complex for full and hence, native T-cell signaling. Here, we tested whether such a gp100(280-288)-or p53(264-272) tumor antigen-specific scTCR is still prone to mispairing with TCR alpha. In a human Jurkat-76 T-cell line lacking endogenous TCRs, surface expression and function of a scTCR could be reconstituted by any co-introduced TCR alpha-chain indicating mispairing to take place on a molecular basis. In contrast, transduction into human TCR alpha/beta-positive T-cells revealed that mispairing is largely reduced. Competition experiments in Jurkat-76 confirmed the preference of dcTCR to selfpair and to spare scTCR. This also allowed for the generation of dc/scTCR-modified cytomegalovirus/tumor antigen-bispecific T-cells to augment T-cell activation in CMV-infected tumor patients. Residual mispairing was prevented by strenghtening the V alpha-Li-V beta-fragment through the design of a novel disulfide bond between a V alpha- and a linker-resident residue close to V beta. Multimer-stainings, and cytotoxicity-, IFN gamma-secretion-, and CFSE-proliferation-assays, the latter towards dendritic cells endogenously processing RNA-electroporated gp100 antigen proved the absence of hybrid scTCR/TCR alpha-formation without impairing avidity of scTCR/C alpha in T-cells. Moreover, a fragile cytomegalovirus pp65(495-503)-specific scTCR modified this way acquired enhanced cytotoxicity. Thus, optimized scTCR/Ca inhibits residual TCR mispairing to accomplish safe adoptive immunotherapy for bulk endogenous TCR alpha/beta-positive T-cells.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | LENTIVIRAL GENE-TRANSFER; VERSUS-HOST-DISEASE; ALPHA-BETA; ANTITUMOR-ACTIVITY; IN-VIVO; RECEPTOR; LYMPHOCYTES; ANTIGEN; THERAPY; COMPLEX; human; tumor Immunity; T-cells; T-cell receptors; gene therapy; Immunology and Microbiology Section; Immune response; Immunity |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 05 Apr 2019 07:08 |
| Last Modified: | 05 Apr 2019 07:08 |
| URI: | https://pred.uni-regensburg.de/id/eprint/3064 |
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