title: Generation of TCR-engineered T cells and their use to control the performance of T cell assays.
abstract: The systematic assessment of the human immune system bears huge potential to guide rational development of novel immunotherapies and clinical decision making. Multiple assays to monitor the quantity, phenotype, and function of Ag-specific T cells are commonly used to unravel patients' immune signatures in various disease settings and during therapeutic interventions. When compared with tests measuring soluble analytes, cellular immune assays have a higher variation, which is a major technical factor limiting their broad adoption in clinical immunology. The key solution may arise from continuous control of assay performance using TCR-engineered reference samples. We developed a simple, stable, robust, and scalable technology to generate reference samples that contain defined numbers of functional Ag-specific T cells. First, we show that RNA-engineered lymphocytes, equipped with selected TCRs, can repetitively deliver functional readouts of a controlled size across multiple assay platforms. We further describe a concept for the application of TCR-engineered reference samples to keep assay performance within or across institutions under tight control. Finally, we provide evidence that these novel control reagents can sensitively detect assay variation resulting from typical sources of error, such as low cell quality, loss of reagent stability, suboptimal hardware settings, or inaccurate gating.
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Nicole
authors.initials: N
authors.lastname: Bidmon
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany; University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Sebastian
authors.initials: S
authors.lastname: Attig
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Richard
authors.initials: R
authors.lastname: Rae
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Helene
authors.initials: H
authors.lastname: Schröder
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Tana A
authors.initials: TA
authors.lastname: Omokoko
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Petra
authors.initials: P
authors.lastname: Simon
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Andreas N
authors.initials: AN
authors.lastname: Kuhn
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kreiter
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany; University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
authors.forename: Ugur
authors.initials: U
authors.lastname: Sahin
authors.affiliation: Department of Immunology, University of Tuebingen, 72076 Tuebingen, Germany; and.
authors.forename: Cécile
authors.initials: C
authors.lastname: Gouttefangeas
authors.affiliation: Department of Clinical Oncology, Leiden University Medical Center, 2333 ZA Leiden, the Netherlands.
authors.forename: Sjoerd H
authors.initials: SH
authors.lastname: van der Burg
authors.affiliation: Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany; cedrik.x.britten@gsk.com.
authors.forename: Cedrik M
authors.initials: CM
authors.lastname: Britten
mesh_terms: D015870:Gene Expression; D005818:Genetic Engineering; D006680:HLA Antigens / Q000737:chemistry / Q000276:immunology; D006801:Humans; D007159:Immunologic Tests / Q000379:methods* / Q000592:standards*; D007167:Immunotherapy / Q000379:methods; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D055503:Protein Multimerization; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000378:metabolism*; D037182:T-Cell Antigen Receptor Specificity / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*
chemical_list: D006680:HLA Antigens; D010455:Peptides; D011948:Receptors, Antigen, T-Cell