Simple SummaryNeutrophil granulocytes are white blood cells that can release so-called neutrophil extracellular traps (NETs) which are composed of DNA and proteins. They help to fight pathogens such as bacteria but can also play a role in cancer. For human melanoma, which is due to its metastases the most lethal skin cancer, it is already known that neutrophils occur in metastases and can lead to a worse survival. Here, we investigate whether NETs can be found within melanoma metastases in order to better understand their role in the immune response to human melanoma and to assess their suitability as a potential therapeutic target. Therefore, we analyzed 81 metastases and found that one half was infiltrated with neutrophils and one third contained NETs. Further, we could see that NETs are predominantly found in large metastases and necrotic areas, i.e., areas with accumulated tumor cell death. NETs were present at all observed sites (skin, lymph node, liver and lung metastases). As many metastases contain NETs, our study delivers a basis to study NETs more intensively in the context of treatment response and the patient’s outcome in melanoma.AbstractNeutrophil extracellular traps (NETs) are web-like structures released by neutrophils that kill invading microorganisms. However, NETs also promote tumor growth and impair the functionality of T-cells in cancer. Therefore, this study aimed at characterizing NET distribution within human melanoma metastases (n = 81 of 60 patients) by immunofluorescence staining for neutrophils (CD15) and NETs (H3Cit) in order to identify targets for NET-directed therapies. The results show that 49.3% of the metastases contained neutrophils (n = 40) and 30.8% (n = 25) contained NETs, 68% of them very densely infiltrated. A total of 75% of CD15-positive neutrophils and 96% of NET-containing metastases were necrotic while metastases without neutrophil infiltration were predominantly non-necrotic. A higher amount of NETs correlated significantly with greater tumor size. Consistently, all metastases with a cross-sectional area greater than 2.1 cm2contained neutrophils. Analysis of metastasis from different sites revealed NETs to be present in skin, lymph node, lung and liver metastases. Taken together, our study was the first to observe NET infiltration in a larger cohort of human melanoma metastases. These results set the stage for further research regarding NET-directed therapies in metastatic melanoma.