Frontiers in Immunology (Oct 2024)

Modified Dendritic cell-based T-cell expansion protocol and single-cell multi-omics allow for the selection of the most expanded and in vitro-effective clonotype via profiling of thousands of MAGE-A3-specific T-cells

  • Sergey Sennikov,
  • Marina Volynets,
  • Saleh Alrhmoun,
  • Roman Perik-Zavodskii,
  • Olga Perik-Zavodskaia,
  • Marina Fisher,
  • Julia Lopatnikova,
  • Julia Shevchenko,
  • Kirill Nazarov,
  • Julia Philippova,
  • Alaa Alsalloum,
  • Vasily Kurilin,
  • Alexander Silkov

DOI
https://doi.org/10.3389/fimmu.2024.1470130
Journal volume & issue
Vol. 15

Abstract

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IntroductionAdoptive cell therapy using TCR-engineered T-cells is one of the most effective strategies against tumor cells. The TCR T-cell approach has been well tested against a variety of blood neoplasms but is yet to be deeply tested against solid tumors. Among solid tumors, cancer-testis antigens are the most prominent targets for tumor-specific therapy, as they are usually found on cells that lie behind blood-tissue barriers.MethodsWe have employed a novel efficient protocol for MAGE-A3-specific T-cell clonal expansion, performed single-cell multi-omic analysis of the expanded T-cells via BD Rhapsody, engineered a selected T-cell receptor into a lentiviral construct, and tested it in an in vitro LDH-cytotoxicity test.Results and discussionWe have observed a 191-fold increase in the MAGE-A3-specific T-cell abundance, obtained a dominant T-cell receptor via single-cell multi-omic BD Rhapsody data analysis in the TCRscape bioinformatics tool, and observed potent cytotoxicity of the dominant-clonotype transduced TCR T-cells against a MAGE-A3-positive tumor. We have demonstrated the efficiency of our T-cell enrichment protocol in obtaining potent anti-tumor T-cells and their T-cell receptors, especially when paired with the modern single-cell analysis methods.

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