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Cited by 155CAR-T cell therapy researchImmunotherapy and Immune ResponsesT-cell and B-cell Immunology

Induction of a central memory and stem cell memory phenotype in functionally active CD4+ and CD8+ CAR T cells produced in an automated good manufacturing practice system for the treatment of CD19+ acute lymphoblastic leukemia

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Blaeschke, Franziska, Stenger, Dana, Kaeuferle, Theresa, Willier, Semjon, Lotfi, Ramin, Kaiser, Andrew Didier, Assenmacher, Mario, Döring, Michaela +2 more·2018·Cancer Immunology, Immunotherapy
DOI 10.1007/s00262-018-2155-7PMC PMC11028239
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Abstract

Relapsed/refractory B-precursor acute lymphoblastic leukemia (pre-B ALL) remains a major therapeutic challenge. Chimeric antigen receptor (CAR) T cells are promising treatment options. Central memory T cells (Tcm) and stem cell-like memory T cells (Tscm) are known to promote sustained proliferation and persistence after T-cell therapy, constituting essential preconditions for treatment efficacy. Therefore, we set up a protocol for anti-CD19 CAR T-cell generation aiming at high Tcm/Tscm numbers. 100 ml peripheral blood from pediatric pre-B ALL patients was processed including CD4+/CD8+-separation, T-cell activation with modified anti-CD3/-CD28 reagents and transduction with a 4-1BB-based second generation CAR lentiviral vector. The process was performed on a closed, automated device requiring additional manual/open steps under clean room conditions. The clinical situation of these critically ill and refractory patients with leukemia leads to inconsistent cellular compositions at start of the procedure including high blast counts and low T-cell numbers with exhausted phenotype. Nevertheless, a robust T-cell product was achieved (mean CD4+ = 50%, CD8+ = 39%, transduction = 27%, Tcm = 50%, Tscm = 46%). Strong proliferative potential (up to > 100-fold), specific cytotoxicity and low expression of co-inhibitory molecules were documented. CAR T cells significantly released TH1 cytokines IFN-γ, TNF-α and IL-2 upon target-recognition. In conclusion, partly automated GMP-generation of CAR T cells from critically small blood samples was feasible with a new stimulation protocol that leads to high functionality and expansion potential, balanced CD4/CD8 ratios and a conversion to a Tcm/Tscm phenotype.

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  • The CliniMACS Prodigy integrates all cell processing steps from T cell separation, CAR transduction, cell expansion, and formulation in a device using a single-use chamber and tubing set under GMP compliance.

    “To this end, we used a closed semi-automated platform, CliniMACS Prodigy, that integrates all cell processing steps from T cell separation, CAR transduction, cell expansion, and formulation in a device using a single-use chamber and tubing set under Good Manufacturing Practices (GMP) compliance.10,11”

    From Point-of-care manufacturing of anti-CD19 CAR-T cells using a closed production platform: Experiences of an academic in Thailand (2024) · cites this paper as [11]

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Research works printed but not linked: 18

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Cited by

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Authors
Blaeschke, F., Stenger, D., Kaeuferle, T., Willier, S., Lotfi, R., Kaiser, A. D., Assenmacher, M., Döring, M., Feucht, J., & Feuchtinger, T.
Affiliations
  • Ludwig-Maximilians-Universität München (DE)
  • Universität Ulm (DE)
  • Miltenyi Biotec (Germany) (DE)
  • University Children's Hospital Tübingen (DE)
  • Memorial Sloan Kettering Cancer Center (US)
  • LMU Klinikum (DE)
Corresponding author
Feuchtinger, Tobias
ORCID
Venue
Cancer Immunology, Immunotherapy
Publisher
Springer Science and Business Media LLC
Published
2018-03-31
Volume
67
Issue
7
Pages
1053-1066
ISSN
  • 0340-7004 (ISSN-L)
  • 1432-0851
DOI
10.1007/s00262-018-2155-7
PMC
PMC11028239
OpenAlex
W2795246883
Cited by
155
References
4
Citations by year
2026: 8 · 2025: 14 · 2024: 23 · 2023: 27 · 2022: 21 · 2021: 24 · 2020: 21 · 2019: 14 · 2018: 2 · 2012: 1
FWCI
5.73
Type
journal article
Language
en
Open access
Licence
www.springer.com/tdm (tdm)
Locations
  • Cancer Immunology Immunotherapy (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Care-for-Rare Foundation
  • Mehr LEBEN für Krebskranke Kinder – Bettina-Bräu-Stiftung
Topics
  • Health Sciences › Medicine › Oncology › CAR-T cell therapy research
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
Keywords
chimeric antigen receptor T cells, central memory T cells, stem cell memory T cells, anti-CD19 CAR, B-cell precursor acute lymphoblastic leukemia, relapsed/refractory leukemia, 4-1BB, lentiviral vectors, T cell activation, T cell exhaustion, cytokine production, pediatric leukemia
MeSH terms
Adolescent, Cytotoxicity, Immunologic, Female, Humans, Immunologic Memory, Lymphocyte Activation, Phenotype, Prognosis, Receptors, Antigen, T-Cell, Stem Cells, CD4-Positive T-Lymphocytes, Cytokines, Immunotherapy, Adoptive, CD8-Positive T-Lymphocytes, Antigens, CD19, Precursor Cell Lymphoblastic Leukemia-Lymphoma

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