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. 2025 Dec;14(1):2546402.
doi: 10.1080/2162402X.2025.2546402. Epub 2025 Aug 15.

Lymphodepletion, tumor-infiltrating lymphocytes, and high versus low dose IL-2 followed by pembrolizumab in patients with metastatic melanoma

Merve Hasanov  1 , Simin Kiany  2 , Marie-Andrée Forget  3 , Roland Bassett  4 , Michael A Davies  5 , Adi Diab  5 , Jeffrey E Gershenwald  6 , Isabella C Glitza  5 , Jeffrey E Lee  6 , Anthony Lucci  6 , Jennifer L McQuade  5 , Sapna P Patel  7 , Merrick I Ross  6 , Hussein A Tawbi  5 , Jennifer A Wargo  6 , Michael K Wong  5 , Chantale Bernatchez  3 , Patrick Hwu  8 , Cara Haymaker  2 , Rodabe N Amaria  5
Affiliations

Lymphodepletion, tumor-infiltrating lymphocytes, and high versus low dose IL-2 followed by pembrolizumab in patients with metastatic melanoma

Merve Hasanov et al. Oncoimmunology. 2025 Dec.

Abstract

This study evaluated the efficacy and safety of unengineered tumor-infiltrating lymphocytes (TILs) combined with pembrolizumab and either high (HD, Arm-1) or low (LD, Arm-2) doses of IL-2 in patients with metastatic melanoma (MM). Patients were lymphodepleted with cyclophosphamide and fludarabine, followed by TIL infusion and IL-2 (Arm-1: 720,000 IU/kg IV q 8 hrs up to 15 doses; Arm-2: 2 million IU SC for 14 days). Patients received pembrolizumab 200 mg IV starting 21 days post-TIL infusion, and every 3 weeks for up to 2 years. The primary endpoint was overall response rate (ORR) per RECIST 1.1. Blood samples were collected for longitudinal flow cytometry and cytokine analysis. In Arm-1 (n = 7), one patient had a partial response (PR) for 10 months, two had stable disease (SD), three had progressive disease (PD), and one was not evaluable (NE). In Arm-2 (n = 7), one patient had an ongoing PR for over 76 months, one had SD, and five had PD. The toxicity profiles were comparable; however, patients in Arm-2 had lower grade 3 febrile neutropenia (57% vs. 71%) and shorter hospitalization (median 16 days vs. 18 days). No correlation was observed between TIL phenotype and clinical response, although PR patients received high numbers of TIL with a high CD8+/CD4+ T cell ratio. IL-2 dose did not affect the frequency, phenotype, or proliferation of circulating T cell subsets, and anti-PD-1 did not boost T-cell proliferation. No significant differences were observed between IL-2 doses, suggesting low-dose IL-2 as an alternative to high-dose IL-2 after TIL administration.

Keywords: Tumor-infiltrating lymphocytes; adoptive cell therapy; anti-PD-1; checkpoint blockade; immune profiling; interleukin 2.

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Conflict of interest statement

MH Advisory board member: BMS.

JEG – Merck – advisory board and scientific advisory committee. S.P.P. reports receiving honoraria from advisory boards, steering committees, data safety monitoring boards, or consulting from Bristol Myers Squibb, Cardinal Health, Castle Biosciences, Ideaya, Immatics, IO Biotech, MSD, Novartis, Obsidian, OncoSec, Pfizer, Replimune, Scancell, and TriSalus Life Sciences.

MAD is supported by Dr. Miriam and Sheldon G. Adelson Medical Research Foundation; AIM at Melanoma Foundation; NIH/NCI P50CA221703; American Cancer Society; Melanoma Research Alliance; Cancer Fighters of Houston, Anne, and John Mendelsohn Chair for Cancer Research; and philanthropic contributions to the Melanoma Moon Shots Program of MD Anderson. MAD has been a consultant for Replimmune, Roche/Genentech, Array, Pfizer, Novartis, BMS, GSK, Sanofi-Aventis, Vaccinex, Apexigen, Eisai, Iovance, Merck, and ABM Therapeutics, and has been the PI of research grants to MD Anderson by Roche/Genentech, GSK, Sanofi-Aventis, Merck, Myriad, Oncothyreon, Pfizer, ABM Therapeutics, and LEAD Pharma.

C.H. declares research funding to institutions from Sanofi, BTG, Iovance, Obsidian, KSQ, EMD Serono, Takeda, Genentech, BMS, Summit Therapeutics, Artidis, Immunogenesis, and Novartis; scientific advisory board member of Briacell with stock options; personal fees from Regeneron outside the scope of the submitted work.

RNA receives research funding from Obsidian, KSQ, Erasca, Roche, OnKure, and Regeneron.

All other authors declare no conflicts of interest related to the study.

Figures

Figure 1.
Figure 1.
No significant difference in progression-free survival and overall survival were detected between the two treatment arms. (A) with a median follow-up of 9.2 months, PFS was 3.9 (95%CI, 2.04-NA) months for Arm-1 and 2.1 (95%CI, 2.04-NA) months for Arm-2 (p = 0.99). (B) median OS was 9.7 (95%CI, 3-NA) months for Arm-1 and 8.8 (8.2-NA) months for Arm-2 (p = 0.71). (C) 6 months, 1- and 5-year progression-free survival (PFS), and overall survival (OS) are shown in panel C in each treatment arm and all patients.
Figure 2.
Figure 2.
IL-2 dose does not play a role in circulating Treg expansion and activation. (A) Treg frequency in circulation in Arm-1 (HD IL-2; red) is not significantly different from Arm-2 (LD IL-2; black) at W3 post-TIL infusion. (B) the expression of activation (ICOS, OX40), proliferation (Ki67) (left graph), or checkpoint receptors (PD-1, CTLA-4, TIGIT, Tim3, LAG-3) (right graph) on Tregs does not differ by Arm (Arm-1, HD IL-2; red and Arm-2, LD IL-2; black). (C) representative flow cytometry dot plots. (D) Ki67 expression on CD4+ non-Treg and CD8+ T cells and their representative dot plots (Arm-1, HD IL-2; red and Arm-2, LD IL-2; black) showing no difference by Arm.
See this image and copyright information in PMC

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