tetano
Editor, Senior Moderator
J Infect Dis
. 2022 Dec 30;jiac500.
doi: 10.1093/infdis/jiac500. Online ahead of print.
Manufacture and Characterization of GMP-Compliant SARS-COV-2 Cytotoxic T Lymphocytes
Yaya Chu[SUP] 1 [/SUP], Jordan Milner[SUP] 1 [/SUP], Margaret Lamb[SUP] 2 3 [/SUP], Elena Maryamchik[SUP] 4 5 [/SUP], Olivia Rigot[SUP] 1 [/SUP], Janet Ayello[SUP] 1 [/SUP], Lauren Harrison[SUP] 1 [/SUP], Rosemarie Shaw[SUP] 1 [/SUP], Gregory K Behbehani[SUP] 6 [/SUP], Elaine R Mardis[SUP] 3 7 [/SUP], Katherine Miller[SUP] 7 [/SUP], Lakshmi Prakruthi Rao Venkata[SUP] 7 [/SUP], Hsiaochi Chang[SUP] 6 [/SUP], Dean Lee[SUP] 2 3 [/SUP], Elana Rosenthal[SUP] 1 [/SUP], Stephan Kadauke[SUP] 4 [/SUP], Nancy Bunin[SUP] 5 8 [/SUP], Julie-An Talano[SUP] 9 [/SUP], Bryon Johnson[SUP] 10 [/SUP], Yongping Wang[SUP] 4 5 [/SUP], Mitchell S Cairo[SUP] 1 11 12 13 [/SUP]
Affiliations
Abstract
Background: Coronavirus disease 2019 (COVID-19) is caused by the 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 IFN-γ+ 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: 93% of convalescent donor blood samples passed the screening criteria for clinical manufacture. Three validation runs resulted in enriched T-cells that were 79% ± 21% IFN-γ+ T-cells. SARS-CoV-2-vCTLs displayed a highly diverse TCR 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.
. 2022 Dec 30;jiac500.
doi: 10.1093/infdis/jiac500. Online ahead of print.
Manufacture and Characterization of GMP-Compliant SARS-COV-2 Cytotoxic T Lymphocytes
Yaya Chu[SUP] 1 [/SUP], Jordan Milner[SUP] 1 [/SUP], Margaret Lamb[SUP] 2 3 [/SUP], Elena Maryamchik[SUP] 4 5 [/SUP], Olivia Rigot[SUP] 1 [/SUP], Janet Ayello[SUP] 1 [/SUP], Lauren Harrison[SUP] 1 [/SUP], Rosemarie Shaw[SUP] 1 [/SUP], Gregory K Behbehani[SUP] 6 [/SUP], Elaine R Mardis[SUP] 3 7 [/SUP], Katherine Miller[SUP] 7 [/SUP], Lakshmi Prakruthi Rao Venkata[SUP] 7 [/SUP], Hsiaochi Chang[SUP] 6 [/SUP], Dean Lee[SUP] 2 3 [/SUP], Elana Rosenthal[SUP] 1 [/SUP], Stephan Kadauke[SUP] 4 [/SUP], Nancy Bunin[SUP] 5 8 [/SUP], Julie-An Talano[SUP] 9 [/SUP], Bryon Johnson[SUP] 10 [/SUP], Yongping Wang[SUP] 4 5 [/SUP], Mitchell S Cairo[SUP] 1 11 12 13 [/SUP]
Affiliations
- PMID: 36583990
- DOI: 10.1093/infdis/jiac500
Abstract
Background: Coronavirus disease 2019 (COVID-19) is caused by the 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 IFN-γ+ 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: 93% of convalescent donor blood samples passed the screening criteria for clinical manufacture. Three validation runs resulted in enriched T-cells that were 79% ± 21% IFN-γ+ T-cells. SARS-CoV-2-vCTLs displayed a highly diverse TCR 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.