PMID- 36583990 OWN - NLM STAT- MEDLINE DCOM- 20230329 LR - 20230403 IS - 1537-6613 (Electronic) IS - 0022-1899 (Linking) VI - 227 IP - 6 DP - 2023 Mar 28 TI - Manufacture and Characterization of Good Manufacturing Practice-Compliant SARS-COV-2 Cytotoxic T Lymphocytes. PG - 788-799 LID - 10.1093/infdis/jiac500 [doi] AB - BACKGROUND: Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 virus-specific cytotoxic T-cell lymphocytes (vCTLs) could provide a promising modality in COVID-19 treatment. We aimed to screen, manufacture, and characterize SARS-CoV-2-vCTLs generated from convalescent COVID-19 donors using the CliniMACS Cytokine Capture System (CCS). METHODS: Donor screening was done by stimulation of convalescent COVID-19 donor peripheral blood mononuclear cells with viral peptides and identification of interferongamma (IFN-gamma)+ CD4 and CD8 T cells using flow cytometry. Clinical-grade SARS-CoV-2-vCTLs were manufactured using the CliniMACS CCS. The enriched SARS-CoV-2-vCTLs were characterized by T-cell receptor sequencing, mass cytometry, and transcriptome analysis. RESULTS: Of the convalescent donor blood samples, 93% passed the screening criteria for clinical manufacture. Three validation runs resulted in enriched T cells that were 79% (standard error of the mean 21%) IFN-gamma+ T cells. SARS-CoV-2-vCTLs displayed a highly diverse T-cell receptor repertoire with enhancement of both memory CD8 and CD4 T cells, especially in CD8 TEM, CD4 TCM, and CD4 TEMRA cell subsets. SARS-CoV-2-vCTLs were polyfunctional with increased gene expression in T-cell function, interleukin, pathogen defense, and tumor necrosis factor superfamily pathways. CONCLUSIONS: Highly functional SARS-CoV-2-vCTLs can be rapidly generated by direct cytokine enrichment (12 hours) from convalescent donors. CLINICAL TRIALS REGISTRATION: NCT04896606. CI - (c) The Author(s) 2022. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. FAU - Chu, Yaya AU - Chu Y AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. FAU - Milner, Jordan AU - Milner J AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. FAU - Lamb, Margaret AU - Lamb M AD - Department of Hematology/Oncology/Bone Marrow Transplant, Center for Childhood Cancer and Blood Diseases, Nationwide Children's Hospital, Columbus, Ohio, USA. AD - Department of Pediatrics, The Ohio State University, Columbus, Ohio, USA. FAU - Maryamchik, Elena AU - Maryamchik E AD - Department of Pathology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA. AD - Perlman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA. FAU - Rigot, Olivia AU - Rigot O AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. FAU - Ayello, Janet AU - Ayello J AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. FAU - Harrison, Lauren AU - Harrison L AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. FAU - Shaw, Rosemarie AU - Shaw R AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. FAU - Behbehani, Gregory K AU - Behbehani GK AD - Department of Internal Medicine, Division of Hematology, The Ohio State University, Columbus, Ohio, USA. FAU - Mardis, Elaine R AU - Mardis ER AD - Department of Pediatrics, The Ohio State University, Columbus, Ohio, USA. AD - The Steve and Cindy Rasmussen Institute for Genomic Medicine, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, USA. FAU - Miller, Katherine AU - Miller K AD - The Steve and Cindy Rasmussen Institute for Genomic Medicine, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, USA. FAU - Prakruthi Rao Venkata, Lakshmi AU - Prakruthi Rao Venkata L AD - The Steve and Cindy Rasmussen Institute for Genomic Medicine, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, USA. FAU - Chang, Hsiaochi AU - Chang H AD - Department of Internal Medicine, Division of Hematology, The Ohio State University, Columbus, Ohio, USA. FAU - Lee, Dean AU - Lee D AD - Department of Hematology/Oncology/Bone Marrow Transplant, Center for Childhood Cancer and Blood Diseases, Nationwide Children's Hospital, Columbus, Ohio, USA. AD - Department of Pediatrics, The Ohio State University, Columbus, Ohio, USA. FAU - Rosenthal, Elana AU - Rosenthal E AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. FAU - Kadauke, Stephan AU - Kadauke S AD - Department of Pathology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA. FAU - Bunin, Nancy AU - Bunin N AD - Perlman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA. AD - Department of Pediatrics, Division of Oncology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA. FAU - Talano, Julie-An AU - Talano JA AD - Department of Pediatrics, Hematology/Oncology and BMT, Children's Hospital of Wisconsin, Medical College of Wisconsin, Milwaukee, Wisconsin, USA. FAU - Johnson, Bryon AU - Johnson B AD - Department of Medicine, Hematology/Oncology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA. FAU - Wang, Yongping AU - Wang Y AD - Department of Pathology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA. AD - Perlman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA. FAU - Cairo, Mitchell S AU - Cairo MS AD - Department of Pediatrics, New York Medical College, Valhalla, New York, USA. AD - Department of Medicine, New York Medical College, Valhalla, New York, USA. AD - Department of Microbiology, Immunology, and Pathology, New York Medical College, Valhalla, New York, USA. AD - Department of Cell Biology and Anatomy, New York Medical College, Valhalla, New York, USA. LA - eng SI - ClinicalTrials.gov/NCT04896606 GR - 1R01FD006363/FD/FDA HHS/United States PT - Journal Article PT - Research Support, U.S. Gov't, P.H.S. PL - United States TA - J Infect Dis JT - The Journal of infectious diseases JID - 0413675 RN - 0 (Cytokines) RN - 82115-62-6 (Interferon-gamma) SB - IM MH - Humans MH - *SARS-CoV-2 MH - T-Lymphocytes, Cytotoxic MH - *COVID-19 MH - Leukocytes, Mononuclear MH - COVID-19 Drug Treatment MH - CD8-Positive T-Lymphocytes MH - CD4-Positive T-Lymphocytes MH - Cytokines MH - Interferon-gamma OTO - NOTNLM OT - TCRB CDR3 repertoire OT - IFN-gamma+ T cells OT - SARS-CoV-2 OT - cytokine capture system OT - immune landscape OT - virus-specific cytotoxic T lymphocytes COIS- Potential conflicts of interest. M. S. C. has served as a consultant for Jazz Pharmaceuticals, Omeros Pharmaceuticals, and Novartis Pharmaceuticals; Speakers Bureau for Jazz Pharmaceuticals, Servier Pharmaceuticals, Amgen, Inc, Sanofi, and Sobi; and Advisory Board for Astra Zeneca. B. J. and S. K. have received research support from Miltenyi Biotec. Y. W. reports the Children's Hospital of Philadelphia Cell and Gene Therapy Laboratory has received free SARS-COV2 peptide reagent for testing. All other authors report no potential conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed. EDAT- 2022/12/31 06:00 MHDA- 2023/03/29 06:04 CRDT- 2022/12/30 13:13 PHST- 2022/08/15 00:00 [received] PHST- 2022/12/29 00:00 [accepted] PHST- 2023/03/29 06:04 [medline] PHST- 2022/12/31 06:00 [pubmed] PHST- 2022/12/30 13:13 [entrez] AID - 6965956 [pii] AID - 10.1093/infdis/jiac500 [doi] PST - ppublish SO - J Infect Dis. 2023 Mar 28;227(6):788-799. doi: 10.1093/infdis/jiac500.