Adoptive cell therapy with autologous tumor infiltrating lymphocytes and low-dose Interleukin-2 in metastatic melanoma patients

Eva Ellebaek, Trine Zeeberg Iversen, Niels Junker, Marco Donia, Lotte Engell-Noerregaard, Özcan Met, Lisbet Rosenkrantz Hölmich, Rikke Sick Andersen, Sine Reker Hadrup, Mads Hald Andersen, Per thor Straten, Inge Marie Svane, Eva Ellebaek, Trine Zeeberg Iversen, Niels Junker, Marco Donia, Lotte Engell-Noerregaard, Özcan Met, Lisbet Rosenkrantz Hölmich, Rikke Sick Andersen, Sine Reker Hadrup, Mads Hald Andersen, Per thor Straten, Inge Marie Svane

Abstract

Background: Adoptive cell therapy may be based on isolation of tumor-specific T cells, e.g. autologous tumor infiltrating lymphocytes (TIL), in vitro activation and expansion and the reinfusion of these cells into patients upon chemotherapy induced lymphodepletion. Together with high-dose interleukin (IL)-2 this treatment has been given to patients with advanced malignant melanoma and impressive response rates but also significant IL-2 associated toxicity have been observed. Here we present data from a feasibility study at a Danish Translational Research Center using TIL adoptive transfer in combination with low-dose subcutaneous IL-2 injections.

Methods: This is a pilot trial (ClinicalTrials.gov identifier: NCT00937625) including patients with metastatic melanoma, PS ≤1, age <70, measurable and progressive disease and no involvement of the central nervous system. Six patients were treated with lymphodepleting chemotherapy, TIL infusion, and 14 days of subcutaneous low-dose IL-2 injections, 2 MIU/day.

Results: Low-dose IL-2 considerably decreased the treatment related toxicity with no grade 3-4 IL-2 related adverse events. Objective clinical responses were seen in 2 of 6 treated patients with ongoing complete responses (30+ and 10+ months), 2 patients had stable disease (4 and 5 months) and 2 patients progressed shortly after treatment. Tumor-reactivity of the infused cells and peripheral lymphocytes before and after therapy were analyzed. Absolute number of tumor specific T cells in the infusion product tended to correlate with clinical response and also, an induction of peripheral tumor reactive T cells was observed for 1 patient in complete remission.

Conclusion: Complete and durable responses were induced after treatment with adoptive cell therapy in combination with low-dose IL-2 which significantly decreased toxicity of this therapy.

Figures

Figure 1
Figure 1
Immune and clinical evaluation of patient 11.a) FACS plot from intracellular cytokine staining showing the percentage of interferon (IFN)-γ and tumor necrosis factor (TNF)-α producing CD3+CD8+T cells after incubation with autologous tumor cell lines or Staphylococcal Enterotoxin B (control). First row showing reactivity of tumor infiltrating lymphocytes (TIL) from the infusion product, second and third row showing reactivity of peripheral blood monocytes (PBMC) 1 week before infusion of TILs and 3 weeks after. b) PET/CT scan from 1 week before infusion of TILs and 8 weeks after infusion of TILs. Arrows outlining the measurable disease.
Figure 2
Figure 2
Infusion product tested for reactivity against a dendritic cell vaccine (patient 11).a) IFN-γ ELIspot analyses; Responses are defined as number of IFN-γ secreting cells per 3 x 103 TILs. b) Intracellular cytokine staining; percentage of T cells staining double positive for IFN-γ and TNF-α (first column) or for CD8 and CD107a (second column) DC-mock: mock-transfected dendritic cell (negative control), vac: vaccine, tri: triple transfected, sur: survivin, hTERT: human Telomerase Reverse Transciptase, TIL: tumor infiltrating lymphocytes.

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