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Advances in preclinical TCR characterization: leveraging cell avidity to identify functional TCRs

  • Andreas Carr ORCID logo , Laura M. Mateyka , Sebastian J. C. Scheu ORCID logo , Ana Bici , Joris Paijmans , Rogier M. Reijmers , Nina Dieminger , Shirin Dildebekova , Noomen Hamed , Karolin Wagner , Dirk H. Busch and Elvira D’Ippolito ORCID logo EMAIL logo
Published/Copyright: April 29, 2024
Biological Chemistry
From the journal Biological Chemistry Volume 405 Issue 7-8

Abstract

T-cell therapy has emerged as an effective approach for treating viral infections and cancers. However, a significant challenge is the selection of T-cell receptors (TCRs) that exhibit the desired functionality. Conventionally in vitro techniques, such as peptide sensitivity measurements and cytotoxicity assays, provide valuable insights into TCR potency but are labor-intensive. In contrast, measuring ligand binding properties (z-Movi technology) could provide an accelerated processing while showing robust correlations with T-cell functions. In this study, we assessed whether cell avidity can predict functionality also in the context of TCR-engineered T cells. To this end, we developed a flexible system for TCR re-expression by generating a Jurkat-derived T cell clone lacking TCR and CD3 expression through CRISPR-Cas9-mediated TRBC knockout. The knockin of a transgenic TCR into the TRAC locus restored TCR/CD3 expression, allowing for CD3-based purification of TCR-engineered T cells. Subsequently, we characterized these engineered cell lines by functional readouts, and assessment of binding properties through the z-Movi technology. Our findings revealed a strong correlation between the cell avidities and functional sensitivities of Jurkat TCR-T cells. Altogether, by integrating cell avidity measurements with our versatile T cell engineering platform, we established an accelerated system for enhancing the in vitro selection of clinically relevant TCRs.

Keywords: cellular avidity; Jurkat cells; T cell engineering; T cell receptor; T cell therapy; TCR functionality
Corresponding author: Elvira D’Ippolito, Institute for Medical Microbiology, Immunology and Hygiene, School of Medicine and Health, Technical University of Munich, Munich, Germany; and German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany, E-mail:
Authors Andreas Carr and Laura Mateyka contributed equally to this work. Authors Dirk H. Busch and Elvira D’Ippolito share co-senior authorship.

Funding source: EIT Health

Award Identifier / Grant number: CoViproteHCt 20877

Funding source: German National Network of University Medicine of the Federal Ministry of Education and Research

Award Identifier / Grant number: NaFoUniMedCovid19, 01KX2021

Funding source: Boehringer Ingelheim Fonds

Funding source: Deutsche Forschungsgesellschaft

Award Identifier / Grant number: SFB-TRR 338/1 2021-452881907

Award Identifier / Grant number: SFB1321/1-329628492

Acknowledgments

We thank Philipp Strobl and Laura Valentiner for their help with experiments. We are also grateful to Domenico Fasci, Trillian Gregg and Andrea Candelli from LUMICKS for the valuable discussions.

  1. Research ethics: Not applicable.

  2. Author contributions: E.D. and D.H.B. conceptualized the study; A.C., L.M.M., E.D. developed methodology; A.C., L.M.M. performed software analyses; A.C., L.M.M., A.B., N.D., S.D., K.W. performed experiments; J.P. and R.M.R. developed methodology and analyzed data; N.H. generated multimers; D.H.B. contributed resources; E.D., A.C. L.M.M. wrote the manuscript; all authors read and approved the manuscript; D.H.B. acquired funding; E.D. and D.H.B. supervised the study and administered the project. E.D. and D.H.B. have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: D.H.B. is co-founder of STAGE Cell Therapeutics GmbH (now Juno Therapeutics/BMS) and T Cell Factory B.V. (now Kite/Gilead). D.H.B. has a consulting contract with and receives sponsored research support from Juno Therapeutics/BMS. J.P. and R.M.R are employed by LUMICKS. All other authors state no conflict of interest.

  4. Research funding: The work was supported by the EIT Health CoViproteHCt 20877, the German National Network of University Medicine of the Federal Ministry of Education and Research (BMBF; NaFoUniMedCovid19, 01KX2021; COVIM), and the Deutsche Forschungsgesellschaft (DFG; German Research Foundation) SFB1321/1-329628492 (project P17) and SFB-TRR 338/1 2021-452881907 (project A01). L.M.M. was supported by a Ph.D. fellowship from the Boehringer Ingelheim Fonds.

  5. Data availability: All data generated within the study are shown in the manuscript. Consumables, models, instruments and software used in this study are listed in Table 1. The raw data can be obtained on request from the corresponding author.

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Received: 2023-11-06
Accepted: 2024-03-28
Published Online: 2024-04-29
Published in Print: 2024-07-26

© 2024 Walter de Gruyter GmbH, Berlin/Boston

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  2. Highlight: New developments in immunoengineering
  3. Highlight: new developments in immunoengineering
  4. Better safe than sorry: dual targeting antibodies for cancer immunotherapy
  5. Bovine ultralong CDR-H3 derived knob paratopes elicit potent TNF-α neutralization and enable the generation of novel adalimumab-based antibody architectures with augmented features
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https://doi.org/10.1515/hsz-2023-0341
Keywords for this article
cellular avidity; Jurkat cells; T cell engineering; T cell receptor; T cell therapy; TCR functionality

Articles in the same Issue

  1. Frontmatter
  2. Highlight: New developments in immunoengineering
  3. Highlight: new developments in immunoengineering
  4. Better safe than sorry: dual targeting antibodies for cancer immunotherapy
  5. Bovine ultralong CDR-H3 derived knob paratopes elicit potent TNF-α neutralization and enable the generation of novel adalimumab-based antibody architectures with augmented features
  6. Development of an enabling platform biotechnology for the production of proteins
  7. Beyond CAR T cells: exploring alternative cell sources for CAR-like cellular therapies
  8. Advances in preclinical TCR characterization: leveraging cell avidity to identify functional TCRs
  9. Unpaired cysteine insertions favor transmembrane dimerization and induce ligand-independent constitutive cytokine receptor signaling
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