PMID- 31568490 OWN - NLM STAT- MEDLINE DCOM- 20200312 LR - 20200312 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 14 IP - 9 DP - 2019 TI - Targeted in-vitro-stimulation reveals highly proliferative multi-virus-specific human central memory T cells as candidates for prophylactic T cell therapy. PG - e0223258 LID - 10.1371/journal.pone.0223258 [doi] LID - e0223258 AB - 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 (TCM) 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 TCM show the same promising features for ACT as their murine counterparts. Using a peptide specific proliferation assay (PSPA) we studied the human Adenovirus- (AdV), Cytomegalovirus- (CMV) and Epstein-Barr virus- (EBV) specific TCM repertoires and determined their functional and proliferative capacities in vitro. TCM 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. TCM cultures showed strong proliferation of a plethora of functional virus-specific T cells. Using PSPA, we could unveil tiniest virus epitope-specific TCM 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 TCM products. Consequently, we conclude that TCM bare high potential for prophylactic low-dose ACT. In addition, use of Fab-Streptamer-purified TCM allows circumventing regulatory restrictions typically found in conventional ACT product generation. These GMP-compatible TCM 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 TCM characterization during concomitant immune monitoring. FAU - Faist, Benjamin AU - Faist B AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen, Munich, Germany. AD - German Center for Infection Research (DZIF), partner site Munich, Munich, Germany. FAU - Schlott, Fabian AU - Schlott F AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen, Munich, Germany. AD - German Center for Infection Research (DZIF), partner site Munich, Munich, Germany. FAU - Stemberger, Christian AU - Stemberger C AD - Juno Therapeutics, Munich, Germany. FAU - Dennehy, Kevin M AU - Dennehy KM AD - German Center for Infection Research (DZIF), partner site Tubingen, Tubingen, Germany. AD - Institute for Medical Virology, University Hospital Tubingen, Tubingen, Germany. FAU - Krackhardt, Angela AU - Krackhardt A AD - Department of Medicine III, Klinikum Rechts der Isar, Technische Universitat Munchen, Munich, Germany. FAU - Verbeek, Mareike AU - Verbeek M AD - Department of Medicine III, Klinikum Rechts der Isar, Technische Universitat Munchen, Munich, Germany. FAU - Grigoleit, Gotz U AU - Grigoleit GU AD - Department of Internal Medicine II, University of Wurzburg, Wuerzburg, Germany. FAU - Schiemann, Matthias AU - Schiemann M AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen, Munich, Germany. FAU - Hoffmann, Dieter AU - Hoffmann D AD - German Center for Infection Research (DZIF), partner site Munich, Munich, Germany. AD - Institute for Virology, Technische Universitat Munchen, Munich, Germany. FAU - Dick, Andrea AU - Dick A AD - Department of Transfusion Medicine and Haemostaseology, Ludwig-Maximilians-Universitat Munchen, Munich, Germany. FAU - Martin, Klaus AU - Martin K AD - Institute of Anaesthesiology, Deutsches Herzzentrum Munchen, Klinik an der Technischen Universitat Munchen, Munich, Germany. FAU - Hildebrandt, Martin AU - Hildebrandt M AD - TUM Cells Interdisciplinary Center for Cellular Therapies, Munich, Germany. FAU - Busch, Dirk H AU - Busch DH AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen, Munich, Germany. AD - German Center for Infection Research (DZIF), partner site Munich, Munich, Germany. FAU - Neuenhahn, Michael AU - Neuenhahn M AUID- ORCID: 0000-0003-0400-2743 AD - Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen, Munich, Germany. AD - German Center for Infection Research (DZIF), partner site Munich, Munich, Germany. AD - TUM Cells Interdisciplinary Center for Cellular Therapies, Munich, Germany. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190930 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Epitopes) RN - 0 (Peptides) SB - IM MH - Adenoviridae/genetics/*immunology MH - Adoptive Transfer MH - Biological Assay MH - CD8-Positive T-Lymphocytes/*immunology/virology MH - Cell Proliferation MH - Cytomegalovirus/genetics/*immunology MH - Epitopes/genetics/*immunology MH - Female MH - Gene Expression MH - Healthy Volunteers MH - Herpesvirus 4, Human/genetics/*immunology MH - Histocompatibility Testing MH - Humans MH - *Immunologic Memory MH - Immunomagnetic Separation/methods MH - Immunophenotyping MH - Lymphocyte Activation MH - Male MH - Peptides/genetics/immunology MH - Primary Cell Culture PMC - PMC6768573 COIS- D.H.B. receives consultancy fees and holds shares from Cellgene. C.S. is an employee of Juno Therapeutics GmbH, CellGene. This does not alter our adherence to PLOS ONE policies on sharing data and materials. All other authors have no relevant conflicts of interests. EDAT- 2019/10/01 06:00 MHDA- 2020/03/13 06:00 PMCR- 2019/09/30 CRDT- 2019/10/01 06:00 PHST- 2019/06/25 00:00 [received] PHST- 2019/09/17 00:00 [accepted] PHST- 2019/10/01 06:00 [entrez] PHST- 2019/10/01 06:00 [pubmed] PHST- 2020/03/13 06:00 [medline] PHST- 2019/09/30 00:00 [pmc-release] AID - PONE-D-19-17923 [pii] AID - 10.1371/journal.pone.0223258 [doi] PST - epublish SO - PLoS One. 2019 Sep 30;14(9):e0223258. doi: 10.1371/journal.pone.0223258. eCollection 2019.