Targeted in-vitro-stimulation reveals highly proliferative multi-virus-specific human central memory T cells as candidates for prophylactic T cell therapy

Faist, Benjamin; Schlott, Fabian; Stemberger, Christian; Dennehy, Kevin M.; Krackhardt, Angela; Verbeek, Mareike; Grigoleit, Götz U.; Schiemann, Matthias; Hoffmann, Dieter; Dick, Andrea; Martin, Klaus; Hildebrandt, Martin; Busch, Dirk H. und Neuenhahn, Michael (2019): Targeted in-vitro-stimulation reveals highly proliferative multi-virus-specific human central memory T cells as candidates for prophylactic T cell therapy.
In: PLOS One 14(9), e0223258

Abstract

Adoptive T cell therapy (ACT) has become a treatment option for viral reactivations in patients undergoing allogeneic hematopoietic stem cell transplantation (alloHSCT). Animal models have shown that pathogen-specific central memory T cells (T-CM) are protective even at low numbers and show long-term survival, extensive proliferation and high plasticity after adoptive transfer. Concomitantly, our own recent clinical data demonstrate that minimal doses of purified (not in-vitro-expanded) human CMV epitope-specific T cells can be sufficient to clear viremia. However, it remains to be determined if human virus-specific T-CM show the same promising features for ACT as their murine counterparts. Using a peptide specific proliferation assay (PSPA) we studied the human Adenovirus- (AdV), Cytomegalo-virus- (CMV) and Epstein-Barr virus- (EBV) specific T-CM repertoires and determined their functional and proliferative capacities in vitro. T-CM products were generated from buffy coats, as well as from non-mobilized and mobilized apheresis products either by flow cytometry-based cell sorting or magnetic cell enrichment using reversible Fab-Streptamers. Adjusted to virus serology and human leukocyte antigen (HLA)-typing, donor samples were analyzed with MHC multimer- and intracellular cytokine staining (ICS) before and after PSPA. T-CM cultures showed strong proliferation of a plethora of functional virus-specific T cells. Using PSPA, we could unveil tiniest virus epitope-specific T-CM populations, which had remained undetectable in conventional ex-vivo-staining. Furthermore, we could confirm these characteristics for mobilized apheresis- and GMP-grade Fab-Streptamer-purified T-CM products. Consequently, we conclude that T-CM bare high potential for prophylactic low-dose ACT. In addition, use of Fab-Streptamer-purified T-CM allows circumventing regulatory restrictions typically found in conventional ACT product generation. These GMP-compatible T-CM can now be used as a broad-spectrum antiviral T cell prophylaxis in alloHSCT patients and PSPA is going to be an indispensable tool for advanced T-CM characterization during concomitant immune monitoring.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 1932-6203
• Sprache: Englisch