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Cited by 15CAR-T cell therapy researchImmunotherapy and Immune ResponsesT-cell and B-cell Immunology

Automated manufacturing and characterization of clinical grade autologous CD20 CAR T cells for the treatment of patients with stage III/IV melanoma

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Aleksandrova, Krasimira, Leise, Jana, Priesner, Christoph, Aktas, Murat, Apel, Michael, Assenmacher, Mario, Bürger, Iris, Richter, Anne +13 more·2024·Frontiers in Immunology
DOI 10.3389/fimmu.2024.1328368PMC PMC11461191
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Abstract

Introduction: Point-of-care (POC) manufacturing of chimeric antigen receptor (CAR) modified T cell has expanded rapidly over the last decade. In addition to the use of CD19 CAR T cells for hematological diseases, there is a growing interest in targeting a variety of tumor-associated epitopes. Methods: Here, we report the manufacturing and characterization of autologous anti-CD20 CAR T cells from melanoma patients within phase I clinical trial (NCT03893019). Using a second-generation lentiviral vector for the production of the CD20 CAR T cells on the CliniMACS Prodigy®. Results: We demonstrated consistency in cell composition and functionality of the products manufactured at two different production sites. The T cell purity was >98.5%, a CD4/CD8 ratio between 2.5 and 5.5 and transduction rate between 34% and 61% on day 12 (harvest). Median expansion rate was 53-fold (range, 42–65-fold) with 1.7-3.8×10 9 CAR T cells at harvest, a sufficient number for the planned dose escalation steps (1×10 5/kg, 1×10 6/kg, 1×10 7/kg BW). Complementary research of some of the products pointed out that the CAR+ cells expressed mainly central memory T-cell phenotype. All tested CAR T cell products were capable to translate into T cell activation upon engagement of CAR target cells, indicated by the increase in pro-inflammatory cytokine release and by the increase in CAR T cell amplification. Notably, there were some interindividual, cell-intrinsic differences at the level of cytokine release and amplification. CAR-mediated T cell activation depended on the level of CAR cognate antigen. Discussion: In conclusion, the CliniMACS Prodigy® platform is well suited for decentralized POC manufacturing of anti-CD20 CAR T cells and may be likewise applicable for the rapid and automated manufacturing of CAR T cells directed against other targets. Clinical trial registration: https://clinicaltrials.gov/study/NCT03893019?cond=Melanoma&term=NCT03893019&rank=1, identifier NCT03893019.

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Authors
Aleksandrova, K., Leise, J., Priesner, C., Aktas, M., Apel, M., Assenmacher, M., Bürger, I., Richter, A., Altefrohne, P., Schubert, C., Holzinger, A., Barden, M., Bezler, V., von Bergwelt-Baildon, M., Borchmann, P., Goudeva, L., Glienke, W., Arseniev, L., Esser, R., Abken, H., & Koehl, U.
Affiliations
  • Medizinische Hochschule Hannover (DE)
  • Miltenyi Biotec (Germany) (DE)
  • Leibniz Institute for Immunotherapy (DE)
  • University Hospital Cologne (DE)
Corresponding author
  • Aleksandrova, Krasimira
  • Leise, Jana
ORCID
Venue
Frontiers in Immunology
Publisher
Frontiers Media SA
Published
2024-09-25
Volume
15
Pages
1328368-1328368
ISSN
1664-3224 (ISSN-L)
DOI
10.3389/fimmu.2024.1328368
PMC
PMC11461191
OpenAlex
W4402849606
Cited by
15
References
40
Citations by year
2026: 7 · 2025: 8
FWCI
2.57
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Immunology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Qucosa (Saxon State and University Library Dresden) (publishedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • Fraunhofer-Publica (Fraunhofer-Gesellschaft) (submittedVersion)
Funders
Medizinischen Hochschule Hannover
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
CAR T-cell therapy, melanoma, point-of-care manufacturing, distributed manufacturing, lentiviral vectors, CliniMACS Prodigy, autologous T cells, T cell expansion, cytokine production, central memory T cells, dose escalation
MeSH terms
Receptors, Chimeric Antigen, Humans, Male, Melanoma, Neoplasm Staging, T-Lymphocytes, Immunotherapy, Adoptive, Antigens, CD20

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