title: Identification of Rare High-Avidity, Tumor-Reactive CD8+ T Cells by Monomeric TCR-Ligand Off-Rates Measurements on Living Cells.
abstract: The avidity of the T-cell receptor (TCR) for antigenic peptides presented by the peptide-MHC (pMHC) on cells is a key parameter for cell-mediated immunity. Yet a fundamental feature of most tumor antigen-specific CD8(+) T cells is that this avidity is low. In this study, we addressed the need to identify and select tumor-specific CD8(+) T cells of highest avidity, which are of the greatest interest for adoptive cell therapy in patients with cancer. To identify these rare cells, we developed a peptide-MHC multimer technology, which uses reversible Ni(2+)-nitrilotriacetic acid histidine tags (NTAmers). NTAmers are highly stable but upon imidazole addition, they decay rapidly to pMHC monomers, allowing flow-cytometric-based measurements of monomeric TCR-pMHC dissociation rates of living CD8(+) T cells on a wide avidity spectrum. We documented strong correlations between NTAmer kinetic results and those obtained by surface plasmon resonance. Using NTAmers that were deficient for CD8 binding to pMHC, we found that CD8 itself stabilized the TCR-pMHC complex, prolonging the dissociation half-life several fold. Notably, our NTAmer technology accurately predicted the function of large panels of tumor-specific T cells that were isolated prospectively from patients with cancer. Overall, our results demonstrated that NTAmers are effective tools to isolate rare high-avidity cytotoxic T cells from patients for use in adoptive therapies for cancer treatment.
authors: Department of Oncology, Lausanne University Hospital Center and University of Lausanne, Lausanne, Switzerland.
authors: Michael
authors: M
authors: Hebeisen
authors: Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland. TCMetrix Sàrl., Epalinges, Switzerland.
authors: Julien
authors: J
authors: Schmidt
authors: Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland. TCMetrix Sàrl., Epalinges, Switzerland.
authors: Philippe
authors: P
authors: Guillaume
authors: Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland.
authors: Petra
authors: P
authors: Baumgaertner
authors: Department of Oncology, Lausanne University Hospital Center and University of Lausanne, Lausanne, Switzerland. Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland.
authors: Daniel E
authors: DE
authors: Speiser
authors: Department of Oncology, Lausanne University Hospital Center and University of Lausanne, Lausanne, Switzerland. Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland. TCMetrix Sàrl., Epalinges, Switzerland.
authors: Immanuel
authors: I
authors: Luescher
authors: Department of Oncology, Lausanne University Hospital Center and University of Lausanne, Lausanne, Switzerland. Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland. Nathalie.Rufer@unil.ch.
authors: Nathalie
authors: N
authors: Rufer
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D006207:Half-Life; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008545:Melanoma / Q000276:immunology; D011485:Protein Binding; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*
chemical_list: D008024:Ligands; D011948:Receptors, Antigen, T-Cell