Frontiers in Immunology (Jun 2022)

Process Development for Adoptive Cell Therapy in Academia: A Pipeline for Clinical-Scale Manufacturing of Multiple TCR-T Cell Products

  • Daniela Nascimento Silva,
  • Michael Chrobok,
  • Giulia Rovesti,
  • Giulia Rovesti,
  • Giulia Rovesti,
  • Katie Healy,
  • Arnika Kathleen Wagner,
  • Panagiota Maravelia,
  • Francesca Gatto,
  • Massimiliano Mazza,
  • Lucia Mazzotti,
  • Volker Lohmann,
  • Margaret Sällberg Chen,
  • Matti Sällberg,
  • Marcus Buggert,
  • Anna Pasetto

DOI
https://doi.org/10.3389/fimmu.2022.896242
Journal volume & issue
Vol. 13

Abstract

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Cellular immunotherapies based on T cell receptor (TCR) transfer are promising approaches for the treatment of cancer and chronic viral infections. The discovery of novel receptors is expanding considerably; however, the clinical development of TCR-T cell therapies still lags. Here we provide a pipeline for process development and clinical-scale manufacturing of TCR-T cells in academia. We utilized two TCRs specific for hepatitis C virus (HCV) as models because of their marked differences in avidity and functional profile in TCR-redirected cells. With our clinical-scale pipeline, we reproduced the functional profile associated with each TCR. Moreover, the two TCR-T cell products demonstrated similar yield, purity, transduction efficiency as well as phenotype. The TCR-T cell products had a highly reproducible yield of over 1.4 × 109 cells, with an average viability of 93%; 97.8–99% of cells were CD3+, of which 47.66 ± 2.02% were CD8+ T cells; the phenotype was markedly associated with central memory (CD62L+CD45RO+) for CD4+ (93.70 ± 5.23%) and CD8+ (94.26 ± 4.04%). The functional assessments in 2D and 3D cell culture assays showed that TCR-T cells mounted a polyfunctional response to the cognate HCV peptide target in tumor cell lines, including killing. Collectively, we report a solid strategy for the efficient large-scale manufacturing of TCR-T cells.

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