Relationship Between Serum Cytokine Profile and Circulating Neutrophils Phenotype in Patients with Benign Ovarian Tumors and Ovarian Cancer

Changes in the cytokine microenvironment determine the anti- (N1) and pro-tumor (N2) activity of neutrophils (Nph), which has been shown in multiple in vitro studies. We aimed to evaluate the relationship between serum cytokine profile and circulating Nph phenotype in patients with benign ovarian tumors (BOT) and ovarian cancer (OC). The study included patients with BOT (n = 15), primary stage I-IV OC patients before treatment (n = 63), and healthy donors (n = 22) as a control group. The levels of cytokines were estimated in Nph and blood serum; NF-kBp65 was assessed in the Nph nuclear fraction. The myeloperoxidase (MPO) activity, the ability to generate ROS (spontaneous NBT test), and phagocytosis were assessed in Nph by the cytochemical method. The CD11b(+), CD16(+), CD64(+), CD95(+ )Nph counts, the neutrophil traps (NETS) generation were assessed by fluorescence microscopy. Atomic force microscopy was used to determine Nph membrane stiffness. Statistical analysis was performed using Statistica 13 (TIBCO, USA). We observed N1 and N2 subpopulations of Nph in patients with BOT and OC, considering the surface markers, cytokine/growth factors expression, and effector functions. Multiple and simple linear regression models allowed us to propose the combinations of serum cytokines which could polarize circulating Nph into pro- and anti-tumor populations, including the cytokines involved in the NF-kB-signaling. According to the logistic regression, analysis could differentiate between OC and BOT. The assessment of circulating N1 and N2 populations may be advantageous to identify the tumor-promoting neutrophil markers and contribute to the development of therapeutic strategies.