ANALYZING LOCAL AND SYSTEMIC IMMUNE RESPONSES IN PATIENTS WITH ACUTE PHLEGMONOUS PERITONITIS USING A FLOW CYTOFLUOROMETRIC METHOD

Kuyun L. О.

CLINICAL AND EXPERIMENTAL MEDICINE

Scentific article

Objective. Appendicitis is one of the common pathologies in surgery and is associated with the development of inflammation at the systemic and local levels. Appendectomy rates worldwide range from 8.9 to 10 cases per 10,000. Acute inflammation of the abdominal cavity is characterized by versatile manifestations of impaired innate and adaptive immunity due to complications of the postoperative period in patients with a diagnosis of acute phlegmonous appendicitis. Object and methods. The study included 23 patients with acute phlegmonous appendicitis at the exacerbation stage with its transition to peritonitis. The control group consisted of individuals without surgical pathology of the abdominal cavity and other inflammatory diseases (15 people). The analysis of PE and peripheral blood (PC) cells was performed on a Beckman Dickinson FC-500 laser flow cytometer from Siemens, USA using Lysis II data acquisition and processing software and View Sonic (HP, USA) computer equipment. The subpopulation composition of leukocytes was also determined using an indirect immunofluorescence test. In addition, the phagocytic activity of monocytes and peritoneal macrophages (PM) was determined by PLC and the morphological method. In parallel, a microbiological study of PE obtained from 23 patients with acute phlegmonous appendicitis was performed. All the results were statistically processed for non-parametric criteria using the Minitab 16 software. The analysis used the KolmogorovSmirnov test and the comparison of two average independent samples by Student’s test. Quantitative changes are presented as mean values and standard deviations. Results and their discussion. The results obtained in this study demonstrated that there were differences in the protective mechanisms of the local and systemic immune responses. The conclusions were drawn based on functions and activity of phagocytic cells, such as macrophages and blood monocytes, as well as the levels of proinflammatory and immunosuppressive cytokines localized accordingly. Most severe inflammations were caused by gram negative and gram positive aerobes, especially in combination with anaerobes, such as Bacteroides. It should be noted, that the levels of proinflammatory and immunosuppressive cytokines varied depending on their localization. In the peritoneal fluid the content of IL-10 changed substantially compared to the same figures in the peripheral blood (164.1±51.2 pg/ ml in the peritoneal fluid and – 143±34.9 pg/ml, p<0.001 in the peripheral blood). The TNF-α levels in the peritoneal fluid increased significantly, whereas proinflammatory cytokine IL1-β and IL-6 levels decreased, however the changes were not significant. Substantial increase in levels of IL-10 in the peritoneal fluid was linked to the high content of pathogens whose antigens in turn exacerbated the synthesis of IL-10 in the abdominal cavity. Significance of the research. The study was conducted in order to determine the characteristics of the local and systemic responses in patients with acute phlegmonous peritonitis during the exacerbation period. One of the methods used on cells of the peritoneal exudate (PE) and peripheral blood cells in the course of the study was a flow cytofluorometric method. Activity of phagocytic cells, peritoneal macrophages, blood monocytes and inflammatory and suppressive cytokine levels of the corresponding localization were studied during the inflammatory stage of acute phlegmonous peritonitis. The type of the pathogen that causes inflammation in the peritoneal cavity was important since it determined the course of the pathological process in the patient’s body. Aerobic bacteria (Escherichia coli, Klebsiella, Staphylococcus aureus) caused most severe inflammation, especially in combination with anaerobes (Bacteroides). In patients who suffered damage to basic stages of phagocytosis, such as the ability to ingest and kill pathogens, their subsequent apoptosis and removal, substantial increase of IL-10 was observed in both the peritoneal cavity and blood serum. This could point to the patient’s transition from peritonitis to sepsis. Further study of phagocytic abnormalities and the way proinflammatory and suppressive cytokines influence the course of inflammation and its exacerbation will in turn allow to develop effective methods of treating phlegmonous peritonitis and return patients to their usual lifestyle.

laser flow cytometer, proinflammatery and immunosuppressive cytokines Il-1β, Il-6, TNF-α, phagocytosis.

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«Bulletin of problems biology and medicine» Issue 4 Part 1 (153), 2019 year, 101-105 pages, index UDK 616.346.2-002.36-06:616.381-002]-097

10.29254/2077-4214-2019-4-1-153-101-105