Platform-based manufacturing of tumor-infiltrating lymphocytes derived from different solid tumor types
16 affiliations
- 1BioTherapeutics Unit, The Netherlands Cancer Institute-Antoni van Leeuwenhoek
- 2Division of Molecular Oncology and Immunology, The Netherlands Cancer Institute-Antoni van Leeuwenhoek
- 3Department of Pharmacy & Pharmacology, The Netherlands Cancer Institute-Antoni van Leeuwenhoek
- 4Department of Research Laboratory for Cell Therapy, Sanquin Blood Supply Foundation
- 5Department of Research T-cell differentiation lab, Sanquin Blood Supply Foundation
- 6Division of Medical Oncology, The Netherlands Cancer Institute-Antoni van Leeuwenhoek
- 7Department of Surgery, The Netherlands Cancer Institute-Antoni van Leeuwenhoek
- 8Department of Pulmonary Disease, Leiden University Medical Center
- 9Department of Gynaecologic Oncology, The Netherlands Cancer Institute-Antoni van Leeuwenhoek
- 10Julius Centre for Health Science and Primary Care, University Medical Center Utrecht
- 11Department of Urology, The Netherlands Cancer Institute-Antoni van Leeuwenhoek
- 12Oncode Institute
- 13Landsteiner Laboratory, Amsterdam Medical Center
- 14Department of Medical Oncology, Leiden University Medical Center
- 15The Melanoma Clinic, Centre Hospitalier Universitaire Vaudois
- 16Department of Pharmaceutical Sciences, Utrecht University
Abstract
Background: In adoptive cell therapy with tumor-infiltrating lymphocytes (TILs), TILs are isolated and expanded ex vivo from an autologous tumor lesion. TIL therapy, comprising preparative lymphodepleting chemotherapy, TIL infusion, and post-infusion supportive interleukin-2 (IL-2) administration, has demonstrated efficacy in patients with metastatic melanoma. Interest in extending TIL therapy to other solid tumors is growing, and a tumor-agnostic manufacturing platform would allow first-in-human trials to be launched rapidly in these indications.
Objective: This study investigated whether a manufacturing process and control strategy developed for metastatic melanoma-derived TILs could be adapted into a tumor-agnostic platform for the generation of TILs from different solid tumor types, referred to as NTU101.
Methods: The feasibility of generating TILs from endometrial carcinoma (ENDC, n=9), non-small cell lung cancer (NSCLC, n=9), and penile cancer (PENC, n=10) lesions was assessed during process development (PD) runs using predefined quality criteria. Throughout PD runs, flow cytometry was used to characterize cells. When available, autologous tumor digest was used to assess the tumor-reactive potential of TILs. Based on PD experience, adjusted quality criteria were applied to manufacture one full-scale GMP NTU101-drug product (DP) per tumor type.
Results: Overall, 89% (n=25/28) of PD NTU101-DPs manufactured met the predefined quality criteria, containing at least 0.5×10^9 and 90% viable CD45+CD3+ cells, less than 1.0% of cellular impurities (viable CD45- and CD19+ cells), and a potency of more than 1.0×10^3 pg/mL IFN-γ secretion upon CD3/CD28-polymer stimulation. Across the tumor types, PD NTU101-DPs had comparable T-cell subset and differentiation state compositions. In vitro tumor-reactive potential was detected in 82% of tested NTU101-DPs (ENDC: 71%, n=5/7; NSCLC: 80%, n= 4/5; and PENC: 100%, n=5/5), although the level of reactivity will need to be confirmed with a more in-depth assay. One GMP NTU101-DP was then manufactured from each tumor type, and all three met NTU101’s quality criteria.
Conclusion: The NTU101 platform produced comparable NTU101-DPs from three solid tumor types beyond melanoma, with in vitro tumor-reactive potential for some batches. This platform can be applied in TIL therapy clinical trials to generate ENDC-, NSCLC-, and PENC-derived TILs, and possibly other solid tumor types.
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BibTeX
@article{lievense2026platformbased,
title = {{Platform-based manufacturing of tumor-infiltrating lymphocytes derived from different solid tumor types}},
author = {Justin Lievense and Liselotte Tas and Maaike van Zon and Kari-Anne Legué-van Duinen and Rosa van Amerongen and Sander Kouwen and Rosalie van Riessen and Femke Sluiter and Jeltsje Hagg and Madelon Badoux and Iris Manders and Eva Vedder and Erica Sellink and Tom Seijkens and Koen Hartemink and Marcus Rijken and Oscar Brouwer and Suzanne Castenmiller and Marten Hansen and Monika Wolkers and John Haanen and Jos Beijnen and Cynthia Nijenhuis and Inge Jedema and Bastiaan Nuijen},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1854761},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1854761/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Platform-based manufacturing of tumor-infiltrating lymphocytes derived from different solid tumor types
AU - Justin Lievense
AU - Liselotte Tas
AU - Maaike van Zon
AU - Kari-Anne Legué-van Duinen
AU - Rosa van Amerongen
AU - Sander Kouwen
AU - Rosalie van Riessen
AU - Femke Sluiter
AU - Jeltsje Hagg
AU - Madelon Badoux
AU - Iris Manders
AU - Eva Vedder
AU - Erica Sellink
AU - Tom Seijkens
AU - Koen Hartemink
AU - Marcus Rijken
AU - Oscar Brouwer
AU - Suzanne Castenmiller
AU - Marten Hansen
AU - Monika Wolkers
AU - John Haanen
AU - Jos Beijnen
AU - Cynthia Nijenhuis
AU - Inge Jedema
AU - Bastiaan Nuijen
PY - 2026
DA - 2026/10/05
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1854761
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1854761/full
AB - Background: In adoptive cell therapy with tumor-infiltrating lymphocytes (TILs), TILs are isolated and expanded ex vivo from an autologous tumor lesion. TIL therapy, comprising preparative lymphodepleting chemotherapy, TIL infusion, and post-infusion supportive interleukin-2 (IL-2) administration, has demonstrated efficacy in patients with metastatic melanoma. Interest in extending TIL therapy to other solid tumors is growing, and a tumor-agnostic manufacturing platform would allow first-in-human trials to be launched rapidly in these indications. Objective: This study investigated whether a manufacturing process and control strategy developed for metastatic melanoma-derived TILs could be adapted into a tumor-agnostic platform for the generation of TILs from different solid tumor types, referred to as NTU101. Methods: The feasibility of generating TILs from endometrial carcinoma (ENDC, n=9), non-small cell lung cancer (NSCLC, n=9), and penile cancer (PENC, n=10) lesions was assessed during process development (PD) runs using predefined quality criteria. Throughout PD runs, flow cytometry was used to characterize cells. When available, autologous tumor digest was used to assess the tumor-reactive potential of TILs. Based on PD experience, adjusted quality criteria were applied to manufacture one full-scale GMP NTU101-drug product (DP) per tumor type. Results: Overall, 89% (n=25/28) of PD NTU101-DPs manufactured met the predefined quality criteria, containing at least 0.5×10^9 and 90% viable CD45+CD3+ cells, less than 1.0% of cellular impurities (viable CD45- and CD19+ cells), and a potency of more than 1.0×10^3 pg/mL IFN-γ secretion upon CD3/CD28-polymer stimulation. Across the tumor types, PD NTU101-DPs had comparable T-cell subset and differentiation state compositions. In vitro tumor-reactive potential was detected in 82% of tested NTU101-DPs (ENDC: 71%, n=5/7; NSCLC: 80%, n= 4/5; and PENC: 100%, n=5/5), although the level of reactivity will need to be confirmed with a more in-depth assay. One GMP NTU101-DP was then manufactured from each tumor type, and all three met NTU101’s quality criteria. Conclusion: The NTU101 platform produced comparable NTU101-DPs from three solid tumor types beyond melanoma, with in vitro tumor-reactive potential for some batches. This platform can be applied in TIL therapy clinical trials to generate ENDC-, NSCLC-, and PENC-derived TILs, and possibly other solid tumor types.
SN - 1664-3224
ER -