Rapid single-cell identification of Epstein-Barr virus-specific T-cell receptors for cellular therapy

Cobo, Maria Fernanda Lammoglia; Welters, Carlotta; Rosenberger, Leonie; Leisegang, Matthias; Dietze, Kerstin; Pircher, Christian; Penter, Livius; Gary, Regina; Bullinger, Lars; Takvorian, Anna; Moosmann, Andreas; Dornmair, Klaus; Blankenstein, Thomas; Kammertons, Thomas; Gerbitz, Armin und Hansmann, Leo (2022): Rapid single-cell identification of Epstein-Barr virus-specific T-cell receptors for cellular therapy. In: Cytotherapy, Bd. 24, Nr. 8: S. 818-826

Abstract

Background and aims: Epstein-Barr virus (EBV) is associated with solid and hematopoietic malignancies. After allogeneic stem cell transplantation, EBV infection or reactivation represents a potentially life-threatening condition with no specific treatment available in clinical routine. In vitro expansion of naturally occurring EBV-specific T cells for adoptive transfer is time-consuming and influenced by the donor's T-cell receptor (TCR) repertoire and requires a specific memory compartment that is non-existent in seronegative individuals. The authors present highly efficient identification of EBV-specific TCRs that can be expressed on human T cells and recognize EBV-infected cells. Methods and Results: Mononuclear cells from six stem cell grafts were expanded in vitro with three HLA- B*35:01-or four HLA-A*02:01-presented peptides derived from six EBV proteins expressed during latent and lytic infection. Epitope-specific T cells expanded on average 42-fold and were single-cell-sorted and TCRab-sequenced. To confirm specificity, 11 HLA-B*35:01-and six HLA-A*02:01-restricted dominant TCRs were expressed on reporter cell lines, and 16 of 17 TCRs recognized their presumed target peptides. To con-firm recognition of virus-infected cells and assess their value for adoptive therapy, three selected HLA- B*35:01-and four HLA-A*02:01-restricted TCRs were expressed on human peripheral blood lymphocytes. All TCR-transduced cells recognized EBV-infected lymphoblastoid cell lines. Conclusions: The authors' approach provides sets of EBV epitope-specific TCRs in two different HLA contexts. Resulting cellular products do not require EBV-seropositive donors, can be adjusted to cell subsets of choice with exactly defined proportions of target-specific T cells, can be tracked in vivo and will help to overcome unmet clinical needs in the treatment and prophylaxis of EBV reactivation and associated malignancies. (C) 2022 International Society for Cell & Gene Therapy. Published by Elsevier Inc.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 1465-3249
• Sprache: Englisch