In vitro 4-1BB stimulation promotes expansion of CD8 + tumor-infiltrating lymphocytes from various sarcoma subtypes
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ORIGINAL ARTICLE
In vitro 4‑1BB stimulation promotes expansion of CD8+ tumor‑infiltrating lymphocytes from various sarcoma subtypes Morten Nielsen1 · Anders Krarup‑Hansen2 · Dorrit Hovgaard3 · Michael Mørk Petersen3 · Anand Chainsukh Loya4 · Marie Christine Wulff Westergaard1 · Inge Marie Svane1 · Niels Junker2 Received: 30 August 2019 / Accepted: 3 April 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Tumor-specific tumor-infiltrating lymphocytes (TILs) can be in vitro expanded and have the ability to induce complete and durable tumor regression in some patients with melanoma following adoptive cell therapy (ACT). In this preclinical study, we investigated the feasibility of expanding TIL from sarcomas, as well as performing functional in vitro analyses on these. TILs were expanded in vitro by the use of IL2 stimulation with or without the addition of 4-1BB and CD3 antibodies. Phenotypical and functional analyses were mainly performed by flow cytometry. TILs were expanded from 25 of 28 (89%) tumor samples from patients with 9 different sarcoma subtypes. TILs were predominantly αβ T-cells of effector memory subtype with CD4+ dominance. In particular, C D8+ TIL highly expressed LAG3 and to a lesser degree PD-1 and BTLA. In total, 10 of 20 TIL cultures demonstrated in vitro recognition of autologous tumor. In some cases, the fraction of tumorreactive T cells was more than 20%. 4-1BB stimulation augmented expansion kinetics and favored C D8+ occurrence. In conclusion, TIL expansion from sarcoma is feasible and expanded TILs highly express LAG3 and comprise multifunctional tumor-reactive T-cells. Keywords Tumor-infiltrating lymphocytes · TIL · Sarcoma · Adoptive cell transfer · T cell expansion · 4-1BB
Inge Marie Svane and Niels Junker have contributed equally to this work. The authors of this paper report on their T cell assays transparently and comprehensively as per field-wide consensus, allowing the community a full understanding and interpretation of presented data as well as a comparison of data between groups. The electronic supplementary materials of this publication include a MIATA checklist. For more details, see miataproject.org. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00262-020-02568-x) contains supplementary material, which is available to authorized users. * Niels Junker [email protected] 1
Department of Oncology, National Center for Cancer Immune Therapy (CCIT‑DK), Copenhagen University Hospital Herlev, Herlev, Denmark
2
Department of Oncology, Copenhagen University Hospital Herlev, Herlev, Denmark
3
Department of Orthopedic Surgery, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
4
Department of Pathology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Abbreviations ACT Adoptive cell therapy BTLA B- and T-lymphocyte attenuator CM Culture medium DMSO Dimethyl sulfoxide FACS Fluorescence-activated cell sorter FTD Fresh tumor digest GIST
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