title: NET-producing CD16highCD62Ldimneutrophils migrate to tumor sites and predict improved survival in patients with HNSCC.
abstract: The concept of functional neutrophil subsets is new and their clinical significance in malignancies is unknown. Our study investigated the role of CD16dimCD62Lhigh, CD16highCD62Lhighand CD16highCD62Ldimneutrophil subsets in head and neck squamous cell carcinoma (HNSCC) patients. These neutrophil subsets may play different roles in immune-related activity in cancer, based on their profile, activation state and migration ability within a tumor site, which may be important in predicting cancer prognoses. Tumor biopsies and blood were obtained from newly diagnosed untreated HNSCC patients and healthy controls. Neutrophil subsets and their phenotype were characterized using flow cytometry. Isolated granulocytes were assessed for anti-tumor immune functions. Compared to controls HNSCC patients exhibited increased CD16highCD62Ldimneutrophils in blood; this subset displayed a distinct phenotypes with high expression of CD11b and CD18. This subset was prone to migrate into the tumor facilitated by tumor-derived IL-8. Furthermore, IL-8 was also found to activate neutrophils and thereby promoting subset transition. Various assays demonstrated that activated CD16highCD62Ldimneutrophils inhibited migration, proliferation and induced apoptosis of FaDu cancer cells. Neutrophil elastase detected in activated CD16highCD62Ldimneutrophils and tumor biopsies suggested that CD16highCD62Ldimneutrophils impart anti-tumoral activity via neutrophil extracellular traps. Furthermore, increased fraction of CD16highCD62Ldimneutrophils was shown to correlate with an increased survival rate. Our study demonstrates the clinical relevance of the CD16highCD62Ldimneutrophil subset, providing evidence for its increased migration capacity, its anti-tumor activity including increased NET formation and finally its correlation with increased survival in HNSCC patients.
authors.affiliation: Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Camilla Rydberg
authors.identifier: 0000-0002-5873-2197
authors.initials: CR
authors.lastname: Millrud
authors.affiliation: Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Åsa
authors.initials: Å
authors.lastname: Kågedal
authors.affiliation: Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Susanna
authors.initials: S
authors.lastname: Kumlien Georén
authors.affiliation: Department of Medicine, Translational Immunology Unit, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Ola
authors.initials: O
authors.lastname: Winqvist
authors.affiliation: Department of Otorhinolaryngology, Lund University, Skåne University Hospital, Malmö, Sweden.
authors.forename: Rolf
authors.initials: R
authors.lastname: Uddman
authors.affiliation: Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Ronia
authors.initials: R
authors.lastname: Razavi
authors.affiliation: Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Eva
authors.initials: E
authors.lastname: Munck-Wikland
authors.affiliation: Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Lars Olaf
authors.initials: LO
authors.lastname: Cardell
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D017209:Apoptosis / Q000502:physiology; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology*; D002465:Cell Movement / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology; D005260:Female; D058851:GPI-Linked Proteins / Q000378:metabolism; D006098:Granulocytes / Q000378:metabolism / Q000473:pathology; D006258:Head and Neck Neoplasms / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D019041:L-Selectin / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000378:metabolism / Q000473:pathology*; D017452:Receptors, IgG / Q000378:metabolism*; D000077195:Squamous Cell Carcinoma of Head and Neck; D015996:Survival Rate
chemical_list: C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; D016209:Interleukin-8; D017452:Receptors, IgG; D019041:L-Selectin
keywords: HNSCC; NETs; anti-tumor immunity; neutrophil phenotype; neutrophil subsets