Combined IL-2, Agonistic CD3 and 4–1BB Stimulation Preserve Clonotype Hierarchy in Propagated Non-Small Cell Lung Cancer Tumor-Infiltrating Lymphocytes

Authors

Parin Shah
Marie-Andrée Forget
Meredith L Frank
Peixin Jiang
Donastas Sakellariou-Thompson
Lorenzo Federico
Roohussaba Khairullah
Chantal Alexia Neutzler
Ignacio Wistuba
Chi-Wan B Chow
Yan Long
Junya Fujimoto
Shiaw-Yih Lin
Anirban Maitra
Marcelo V Negrao
Kyle Gregory Mitchell
Annikka Weissferdt
Ara A Vaporciyan
Tina Cascone
Jack A Roth
Jianjun Zhang
Boris Sepesi
Don L Gibbons
John V Heymach
Cara L Haymaker
Daniel J McGrail
Alexandre Reuben
Chantale Bernatchez

Publication Date

2-1-2022

Journal

Journal of Immunotherapy of Cancer

Abstract

Background: Adoptive cell transfer (ACT) of tumor-infiltrating lymphocytes (TIL) yielded clinical benefit in patients with checkpoint blockade immunotherapy-refractory non-small cell lung cancer (NSCLC) prompting a renewed interest in TIL-ACT. This preclinical study explores the feasibility of producing a NSCLC TIL product with sufficient numbers and enhanced attributes using an improved culture method.

Methods: TIL from resected NSCLC tumors were initially cultured using (1) the traditional method using interleukin (IL)-2 alone in 24-well plates (TIL 1.0) or (2) IL-2 in combination with agonistic antibodies against CD3 and 4-1BB (Urelumab) in a G-Rex flask (TIL 3.0). TIL subsequently underwent a rapid expansion protocol (REP) with anti-CD3. Before and after the REP, expanded TIL were phenotyped and the complementarity-determining region 3 β variable region of the T-cell receptor (TCR) was sequenced to assess the T-cell repertoire.

Results: TIL 3.0 robustly expanded NSCLC TIL while enriching for CD8+ TIL in a shorter manufacturing time when compared with the traditional TIL 1.0 method, achieving a higher success rate and producing 5.3-fold more TIL per successful expansion. The higher proliferative capacity and CD8 content of TIL 3.0 was also observed after the REP. Both steps of expansion did not terminally differentiate/exhaust the TIL but a lesser differentiated population was observed after the first step. TIL initially expanded with the 3.0 method exhibited higher breadth of clonotypes than TIL 1.0 corresponding to a higher repertoire homology with the original tumor, including a higher proportion of the top 10 most prevalent clones from the tumor. TIL 3.0 also retained a higher proportion of putative tumor-specific TCR when compared with TIL 1.0. Numerical expansion of TIL in a REP was found to perturb the clonal hierarchy and lessen the proportion of putative tumor-specific TIL from the TIL 3.0 process.

Conclusions: We report the feasibility of robustly expanding a T-cell repertoire recapitulating the clonal hierarchy of the T cells in the NSCLC tumor, including a large number of putative tumor-specific TIL clones, using the TIL 3.0 methodology. If scaled up and employed as a sole expansion platform, the robustness and speed of TIL 3.0 may facilitate the testing of TIL-ACT approaches in NSCLC.

Keywords

Aged, Aged, 80 and over, CD3 Complex, Carcinoma, Non-Small-Cell Lung, Female, Humans, Interleukin-2, Lung Neoplasms, Lymphocytes, Tumor-Infiltrating, Male, Middle Aged, Translational Research, Biomedical, Tumor Necrosis Factor Receptor Superfamily, Member 9, immunotherapy, adoptive, lung neoplasms, lymphocytes, tumor-infiltrating, translational medical research, receptors, antigen

DOI

10.1136/jitc-2021-003082

PMID

35110355

PMCID

PMC8811607

PubMedCentral® Posted Date

2-2-2022

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes