Impaired ex vivo T-cell expansion before leukapheresis is associated with out-of-specification tisagenlecleucel manufacturing in pediatric CD19+ALL
6 affiliations
- 1Department of Pediatrics, Goethe-University Frankfurt
- 2Department of Hematology and Oncology, University Medical Center of the Johannes Gutenberg University
- 3Department of Pediatric Hematology/Oncology and BMT, Wroclaw Medical University
- 4Department of Pediatric Oncology, Hematology and Clinical Immunology, Division of Pediatric Stem Cell Therapy, Medical Faculty, Heinrich Heine University
- 5Pediatric Hematology and Oncology, Hannover Medical School
- 6Department of Pediatric Hematology and Oncology, University Children’s Hospital Münster
Abstract
Introduction: Chimeric antigen receptor (CAR)-T cell therapy requires individual production for each patient, starting with a heterogeneous apheresis product of variable cellular composition and function. This inter-patient variability presents a major challenge for standardizing manufacturing processes and may lead to delays or even production failure. The primary aim of this study was to identify pre-leukapheresis characteristics associated with subsequent manufacturing deviations.
Methods: Multiple T cell-related parameters including T cell receptor (TCR) Vβ repertoire, differentiation status, proliferation capacity (e.g. fraction diluted – DIL), and exhaustion were examined in n = 27 patients before leukapheresis for Kymriah ® immunotherapy. These parameters were analyzed for associations with manufacturing specification compliance and out-of-specification (OOS) events.
Results: Impaired T-cell proliferation was the parameter most strongly associated with manufacturing specification non-conformance. Six of 21 evaluable patients showed poor proliferation (DIL <80%), including all four patients with OOS manufacturing outcomes. In contrast, all remaining patients had DIL values ≥84.9%. Exploratory associations between selected TCR Vβ subsets and manufacturing specifications were also identified.
Conclusion: DIL was the parameter most strongly associated with manufacturing specification compliance and may represent a simple functional measure of pre-apheresis T-cell fitness.
Related
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- Immunomodulatory strategies for ex vivo immune cell expansion in adoptive cell therapy: translational and regulatory considerations for clinical manufacturing
- Engineering ex vivo T-cell expansion for CAR-T therapy: bridging biological fidelity and bioprocess scalability through platforms, functional consumables, and integrated closed manufacturing
- An Analysis of Four Patients Discontinuing Idecabtagene Vicleucel after Leukapheresis due to Multiple Myeloma Progression
- Long-Term Clinical Outcomes of Tisagenlecleucel in Pediatric and Young Adult Patients With Relapsed/Refractory ALL
BibTeX
@article{bremm2026impaired,
title = {{Impaired ex vivo T-cell expansion before leukapheresis is associated with out-of-specification tisagenlecleucel manufacturing in pediatric CD19+ALL}},
author = {Melanie Bremm and Greta Heinze and Lara Schneeberger and Emilia Salzmann-Manrique and Julia Banisharif Dehkordi and Andreas Peinelt and Krzysztof Kalwak and Monika Mielcarek-Siedziuk and Roland Meisel and Rita Beier and Claudia Rossig and Andrea Jarisch and Juliane Schlueter and Eva Rettinger and Jan-Henning Klusmann and Claudia Cappel and Peter Bader and Sabine Huenecke},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1973579},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1973579/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Impaired ex vivo T-cell expansion before leukapheresis is associated with out-of-specification tisagenlecleucel manufacturing in pediatric CD19+ALL
AU - Melanie Bremm
AU - Greta Heinze
AU - Lara Schneeberger
AU - Emilia Salzmann-Manrique
AU - Julia Banisharif Dehkordi
AU - Andreas Peinelt
AU - Krzysztof Kalwak
AU - Monika Mielcarek-Siedziuk
AU - Roland Meisel
AU - Rita Beier
AU - Claudia Rossig
AU - Andrea Jarisch
AU - Juliane Schlueter
AU - Eva Rettinger
AU - Jan-Henning Klusmann
AU - Claudia Cappel
AU - Peter Bader
AU - Sabine Huenecke
PY - 2026
DA - 2026/10/27
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1973579
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1973579/full
AB - Introduction: Chimeric antigen receptor (CAR)-T cell therapy requires individual production for each patient, starting with a heterogeneous apheresis product of variable cellular composition and function. This inter-patient variability presents a major challenge for standardizing manufacturing processes and may lead to delays or even production failure. The primary aim of this study was to identify pre-leukapheresis characteristics associated with subsequent manufacturing deviations. Methods: Multiple T cell-related parameters including T cell receptor (TCR) Vβ repertoire, differentiation status, proliferation capacity (e.g. fraction diluted – DIL), and exhaustion were examined in n = 27 patients before leukapheresis for Kymriah ® immunotherapy. These parameters were analyzed for associations with manufacturing specification compliance and out-of-specification (OOS) events. Results: Impaired T-cell proliferation was the parameter most strongly associated with manufacturing specification non-conformance. Six of 21 evaluable patients showed poor proliferation (DIL <80%), including all four patients with OOS manufacturing outcomes. In contrast, all remaining patients had DIL values ≥84.9%. Exploratory associations between selected TCR Vβ subsets and manufacturing specifications were also identified. Conclusion: DIL was the parameter most strongly associated with manufacturing specification compliance and may represent a simple functional measure of pre-apheresis T-cell fitness.
SN - 1664-3224
ER -