Influence of peritoneal fluid environment from female patients with endometriosis on migration and differentiation of Treg and Th17 lymphocytes

Endometriosis is a common, chronic gynecological disease, manifested by the presence of endomeric tissue outside the uterus, most often in the smaller pelvis. The disease is accompanied by chronic pain and it is one of the main causes of infertility, which makes it a significant clinical and social problem. The etiology of endometriosis is unclear, and the underlying pathological mechanisms are still poorly understood. The presence of endometriosis foci is most likely the cause of the induction of chronic inflammatory reactions manifested by the accumulation of activated macrophages and lymphocytes and the secretion of large amounts of various pro-inflammatory cytokines. Endometriosis is also associated with autoimmune disorders and therefore some consider it an autoimmune disease. It has been suggested that endometriosis and the associated immunopathogenetic phenomena may be caused by a disturbance in the distribution and activation of the Treg and Th17 lymphocyte systems. Treg cells perform suppressor functions, are responsible for the phenomenon of auto-tolerance and inhibit inflammatory and autoimmune reactions, while Th17 cells are responsible for stimulating immune responses, especially inflammatory and autoimmune responses. Our previous observations show that endometriosis is associated with an increase in the number of Treg cells in the peritoneal fluid, which may be important for the course of the immune phenomena underlying this disease. MHowever, the mechanism responsible for this phenomenon remains unknown. The role that Th17 cells may play in endometriosis is also unknown. We hypothesize that the disturbance of Treg cell distribution and, possibly, Th17 cells is associated with chemotaxis and differentiation of these cells under the influence of the cytokine-rich peritoneal fluid environment. Therefore, the aim of our project is to investigate the influence of the peritoneal fluid environment of patients with endometriosis on chemotaxis and differentiation of Treg and Th17 cells, and to determine if there is a correlation between the level of specific cytokines / chemokines in the peritoneal fluid and the occurrence, migration and differentiation of these cells. The research will be conducted in close cooperation with the 1st Department of Obstetrics and Gynecology at the Medical University of Warsaw. The material used for the research will be the peritoneal fluid obtained during the laparoscopic procedure and peripheral blood from about 30 patients with endometriosis and a similar number of patients qualified for the control group. In the peritoneal fluid, the concentration of selected cytokines / chemokines will be determined by ELISA tests. The blood and peritoneal fluid subpopulations of Treg and Th17 will be determined by flow cytometry. The chemotactic and stimulating activity of the peritoneal fluid will be determined by in vitro assays using isolated Treg cells and a CD4 + cell population containing Treg and Th17 precursor cells. These cells will be isolated from buffy coat by immunomagnetic or flow cytometer (FACS Aria) sorting. Individual patient test results will be analyzed using multiple linear regression and logistic regression models. We believe that our proposed research will contribute to deepening the knowledge about the causes and mechanisms of Treg and Th17 cell system disorders in endometriosis. This knowledge may be useful in creating the concept of new methods of therapy for this frequent, but still mysterious, disease,