CD8+ chimeric antigen receptor T cells manufactured in absence of CD4+ cells exhibit hypofunctional phenotype
© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ..
BACKGROUND: Cell culture conditions during manufacturing can impact the clinical efficacy of chimeric antigen receptor (CAR) T cell products. Production methods have not been standardized because the optimal approach remains unknown. Separate CD4+ and CD8+ cultures offer a potential advantage but complicate manufacturing and may affect cell expansion and function. In a phase 1/2 clinical trial, we observed poor expansion of separate CD8+ cell cultures and hypothesized that coculture of CD4+ cells and CD8+ cells at a defined ratio at culture initiation would enhance CD8+ cell expansion and simplify manufacturing.
METHODS: We generated CAR T cells either as separate CD4+ and CD8+ cells, or as combined cultures mixed in defined CD4:CD8 ratios at culture initiation. We assessed CAR T cell expansion, phenotype, function, gene expression, and in vivo activity of CAR T cells and compared these between separately expanded or mixed CAR T cell cultures.
RESULTS: We found that the coculture of CD8+ CAR T cells with CD4+ cells markedly improves CD8+ cell expansion, and further discovered that CD8+ cells cultured in isolation exhibit a hypofunctional phenotype and transcriptional signature compared with those in mixed cultures with CD4+ cells. Cocultured CAR T cells also confer superior antitumor activity in vivo compared with separately expanded cells. The positive impact of CD4+ cells on CD8+ cells was mediated through both cytokines and direct cell contact, including CD40L-CD40 and CD70-CD27 interactions.
CONCLUSIONS: Our data indicate that CD4+ cell help during cell culture maintains robust CD8+ CAR T cell function, with implications for clinical cell manufacturing.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:11 |
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| Enthalten in: |
Journal for immunotherapy of cancer - 11(2023), 11 vom: 20. Nov. |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Lee, Sang Yun [VerfasserIn] |
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| Links: |
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| Themen: |
CD4-CD8 Ratio |
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| Anmerkungen: |
Date Completed 23.01.2024 Date Revised 15.03.2024 published: Electronic Citation Status MEDLINE |
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| doi: |
10.1136/jitc-2023-007803 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM367421909 |
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| 100 | 1 | |a Lee, Sang Yun |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a CD8+ chimeric antigen receptor T cells manufactured in absence of CD4+ cells exhibit hypofunctional phenotype |
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| 500 | |a Date Completed 23.01.2024 | ||
| 500 | |a Date Revised 15.03.2024 | ||
| 500 | |a published: Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ. | ||
| 520 | |a BACKGROUND: Cell culture conditions during manufacturing can impact the clinical efficacy of chimeric antigen receptor (CAR) T cell products. Production methods have not been standardized because the optimal approach remains unknown. Separate CD4+ and CD8+ cultures offer a potential advantage but complicate manufacturing and may affect cell expansion and function. In a phase 1/2 clinical trial, we observed poor expansion of separate CD8+ cell cultures and hypothesized that coculture of CD4+ cells and CD8+ cells at a defined ratio at culture initiation would enhance CD8+ cell expansion and simplify manufacturing | ||
| 520 | |a METHODS: We generated CAR T cells either as separate CD4+ and CD8+ cells, or as combined cultures mixed in defined CD4:CD8 ratios at culture initiation. We assessed CAR T cell expansion, phenotype, function, gene expression, and in vivo activity of CAR T cells and compared these between separately expanded or mixed CAR T cell cultures | ||
| 520 | |a RESULTS: We found that the coculture of CD8+ CAR T cells with CD4+ cells markedly improves CD8+ cell expansion, and further discovered that CD8+ cells cultured in isolation exhibit a hypofunctional phenotype and transcriptional signature compared with those in mixed cultures with CD4+ cells. Cocultured CAR T cells also confer superior antitumor activity in vivo compared with separately expanded cells. The positive impact of CD4+ cells on CD8+ cells was mediated through both cytokines and direct cell contact, including CD40L-CD40 and CD70-CD27 interactions | ||
| 520 | |a CONCLUSIONS: Our data indicate that CD4+ cell help during cell culture maintains robust CD8+ CAR T cell function, with implications for clinical cell manufacturing | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a CD4-CD8 Ratio | |
| 650 | 4 | |a CD8-Positive T-Lymphocytes | |
| 650 | 4 | |a Cell Engineering | |
| 650 | 4 | |a Immunotherapy, Adoptive | |
| 650 | 4 | |a Receptors, Chimeric Antigen | |
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| 700 | 1 | |a Lee, Dong Hoon |e verfasserin |4 aut | |
| 700 | 1 | |a Sun, Wei |e verfasserin |4 aut | |
| 700 | 1 | |a Cervantes-Contreras, Francisco |e verfasserin |4 aut | |
| 700 | 1 | |a Basom, Ryan S |e verfasserin |4 aut | |
| 700 | 1 | |a Wu, Feinan |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Si |e verfasserin |4 aut | |
| 700 | 1 | |a Rai, Richa |e verfasserin |4 aut | |
| 700 | 1 | |a Mirzaei, Hamid R |e verfasserin |4 aut | |
| 700 | 1 | |a O'Steen, Shyril |e verfasserin |4 aut | |
| 700 | 1 | |a Green, Damian J |e verfasserin |4 aut | |
| 700 | 1 | |a Shadman, Mazyar |e verfasserin |4 aut | |
| 700 | 1 | |a Till, Brian G |e verfasserin |4 aut | |
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