Abstract Purpose Immune checkpoint inhibitors can induce a T cell–mediated anti-tumor immune response in patients with melanoma. Visualizing T cell activity using positron emission tomography (PET) might allow early insight into treatment efficacy. Activated tumor–infiltrating T cells express the high-affinity interleukin-2 receptor (IL-2R). Therefore, we performed a pilot study, using fluorine-18-labeled IL-2 ([¹⁸F]FB-IL2 PET), to evaluate whether a treatment-induced immune response can be detected. Methods Patients with metastatic melanoma received ~ 200 MBq [¹⁸F]FB-IL2 intravenously, followed by a PET/CT scan before and during immune checkpoint inhibitor therapy. [¹⁸F]FB-IL2 uptake was measured as standardized uptake value in healthy tissues (SUV_mean) and tumor lesions (SUV_max). Response to therapy was assessed using RECIST v1.1. Archival tumor tissues were used for immunohistochemical analyses of T cell infiltration. Results Baseline [¹⁸F]FB-IL2 PET scans were performed in 13 patients. SUV_mean at baseline was highest in the kidneys (14.2, IQR: 11.6–18.0) and liver (10.6, IQR: 8.6–13.4). In lymphoid tissues, uptake was highest in spleen (10.9, IQR: 8.8–12.4) and bone marrow (2.5, IQR: 2.1–3.0). SUV_max in tumor lesions (n = 41) at baseline was 1.9 (IQR: 1.7–2.3). In 11 patients, serial imaging was performed, three at week 6, seven at week 2, and one at week 4. Median [¹⁸F]FB-IL2 tumor uptake decreased from 1.8 (IQR: 1.7–2.1) at baseline to 1.7 (IQR: 1.4–2.1) during treatment (p = 0.043). Changes in [¹⁸F]FB-IL2 tumor uptake did not correlate with response. IL-2R expression in four archival tumor tissues was low and did not correlate with baseline [¹⁸F]FB-IL2 uptake. No [¹⁸F]FB-IL2-related side effects occurred. Conclusion PET imaging of the IL-2R, using [¹⁸F]FB-IL2, is safe and feasible. In this small patient group, serial [¹⁸F]FB-IL2-PET imaging did not detect a treatment-related immune response. Trial registration Clinicaltrials.gov: NCT02922283; EudraCT: 2014-003387.20