INDICATORS OF INNATE IMMUNITY IN PATIENTS WITH ABDOMINAL SEPSIS WITH DIFFERENT OUTCOMES



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Abstract

Abstract

Innate immunity plays a critical role in early recognition and response to pathogens, and its study in abdominal sepsis (AS) is important for pathogenesis, early diagnosis, prognosis, and therapeutic strategy.

The aim is to study the indicators of innate immunity in the dynamics of AU with different outcomes.

Materials and methods: The study was conducted on 64 patients with AS aged 32-82 years, who were divided into groups: 1 (n = 46) with a favorable outcome and group 2 (n = 18) with a fatal outcome. The studies were conducted on days 1, 3, and 7. The total number of leukocytes was examined using a Sysmex XT – 1800i / XT – 2000i hematology analyzer (Japan), and the functional activity of neutrophils was measured by phagocytosis activity and intensity, and oxygen–dependent metabolism. The program "Statistica 10.0 for Windows" was used for statistical research.

Research results and conclusions. It was found that in patients with AS, the number of metamyelocytes, rod-shaped, segmented neutrophils increases in the blood over the course of 1-7 days of follow-up, regardless of the outcome; with an unfavorable outcome of AS, neutrophilic leukocytosis is more pronounced, and the number of monocytes and eosinophils increases. In patients with AS, regardless of the outcome, the absorption capacity of blood neutrophils increases, oxygen-dependent metabolism in a spontaneous mode, and oxygen-dependent metabolism in an induced mode is inhibited.; with an unfavorable outcome of AS, an escalation in the growth of the absorption capacity of blood neutrophils is observed on days 1-7 of follow-up, against the background of inhibition of oxygen-dependent neutrophil metabolism in spontaneous and induced modes, mainly on days 3 and 7 of follow-up.

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Introduction. Abdominal sepsis (AS) is a syndrome based on a systemic inflammatory response of the body to an intra-abdominal infection, leading to acute organ dysfunction; mortality in AS ranges from 7.6 to 36% [5, 6]. Innate immunity plays a critical role in early recognition and response to pathogens, and its study in AU is important for understanding pathogenesis, early diagnosis and prediction of outcomes, and choosing a therapeutic strategy. The aim is to study the indicators of innate immunity in the dynamics of AU with different outcomes.
Methods. A cross-section using a continuous sampling method was performed as patients with abdominal surgical pathology were hospitalized in the intensive care unit of the State Medical Institution of the Russian Academy of Medical Sciences "GKB No. 8 Chelyabinsk", who underwent preliminary surgical treatment with sanitation of the primary focus during the first day of hospitalization. All patients were diagnosed with sepsis according to the Sepsis-3 concept [10, 11]. The study was conducted on 64 patients with AS aged 32-82 years in accordance with the Helsinki Declaration of the World Medical Association "Ethical Principles of Scientific and Medical Research involving humans" dated 1964 with additions in 2013 and approved by the Ethics Committee of the Federal State Budgetary Educational Institution of Higher Medical Education of the Ministry of Health of the Russian Federation (Protocol No. 10 dated 02.11.2023). In accordance with the outcome of AS group 1 (n = 46) with a favorable outcome and group 2 (n = 18) with a fatal outcome were identified. The studies were conducted on days 1, 3, and 7. The Sysmex XT – 1800i / XT – 2000i hematology analyzer (Japan) was used to determine the total number of leukocytes. Neutrophils were isolated on a ficoll-urographin density gradient and their functional activity was assessed by the activity and intensity of phagocytosis, oxygen-dependent metabolism (activity and intensity of spontaneous and induced HCT tests) [2]. Statistical data processing was performed using the software package "Statistica 10.0 for Windows". The differences at p<0.05 were considered statistically significant.
Results. 
In patients with a favorable outcome of AS, a study of the leukocyte formula compared with a group of healthy individuals revealed an increase in the total number of leukocytes on days 1, 3, and 7 due to rod-shaped and segmented neutrophils, as well as young neutrophils on days 1 and 3 (Table 1). In the dynamics of observation, the number of juvenile neutrophils on day 3 is less than on day 1, on days 7 is less than on days 1 and 3; the number of rod-shaped neutrophils on days 3 and 7 is less than on days 1; the number of eosinophils on days 3 and 7 is more than on days 1. In patients with AS with an unfavorable outcome, a study of the quantitative composition of leukocytes in the blood compared with a group of healthy individuals revealed an increase in the total number of leukocytes on days 1, 3, and 7 due to juvenile, rod-shaped, and segmented neutrophils, and monocytes on days 1 and 7. In the dynamics of observation, it was recorded that the total number of neutrophils, including segmented neutrophils, was 3 days less than 1 day, 7 days more than 1 and 3 days; the number of young neutrophils was 3 days less than 1 day, 7 days more than 3 days; the number of rod-shaped neutrophils and monocytes on day 3 is less than on day 1, on day 7 is more than on day 3; the number of eosinophils on day 7 is less than on days 1 and 3. A comparative characteristic of the quantitative composition of leukocytes in groups 1 and 2 showed that in patients with an unfavorable outcome of AS, in contrast to the group of patients with a favorable outcome of AS, the total number of leukocytes is higher due to juvenile, rod-shaped and segmented neutrophils on days 1, 3 and 7; monocytes and eosinophils – on day 1.
Evaluation of the functional activity of blood neutrophils in the group of patients with a favorable outcome of AS compared with the group of healthy individuals revealed a decrease in phagocytosis activity on day 1, an increase in phagocytosis intensity on day 3, an increase in spontaneous HCT test activity on days 1 and 3, an increase in spontaneous HCT test intensity on day 3, a decrease in induced HCT activity -tests on days 3 and 7, a decrease in the intensity of the induced HCT test on days 1 and 7 (Table 2). The dynamics of the observation revealed that the activity of neutrophil phagocytosis was higher on days 3 and 7 than on day 1.; the intensity of neutrophil phagocytosis and the intensity of the induced HCT test on day 3 are higher than on day 1, on day 7 higher than on day 1, lower than on day 3; the activity of the spontaneous HCT test on day 3 is higher than on day 1, on day 7 lower than on day 1 and Day 3; the intensity of the spontaneous HCT test on day 3 is higher than on day 1, on day 7 is higher than on day 1, lower than on day 3; the activity of the induced HCT test on day 3 is lower than on day 1, on day 7 is lower than on day 1 and 3 a day.
When assessing the functional activity of blood neutrophils in patients with an unfavorable outcome of AS compared with the group of healthy patients, an increase in the activity and intensity of phagocytosis on days 1, 3 and 7, an increase in the activity of spontaneous HCT test on days 1 and 3, a decrease in the intensity of spontaneous HCT test on day 1 and an increase on day 3, a decrease in the activity of the induced HCT test on day 7, the intensity of the induced HCT test on day 1. In the dynamics of observation, it was recorded that the activity of neutrophil phagocytosis on day 3 is lower than on day 1, on day 7 is lower than on day 1, but higher than on day 3; the intensity of neutrophil phagocytosis on day 3 is higher than on day 1, on day 7 is higher than on day 1, but lower than on day 3; the activity of spontaneous and induced HCT tests is higher on day 3 than on day 1, and lower on day 7 than on days 1 and 3; The intensity of the spontaneous HCT test on day 3 is higher than on day 1, on day 7 higher than on day 1, lower than on day 3, and the intensity of the induced HCT test on days 3 and 7 is higher than on day 1.
A comparative characteristic of the functional activity of blood neutrophils in groups 1 and 2 revealed that in patients with an unfavorable outcome of AS, unlike the group with a favorable outcome of AS, the activity of neutrophil phagocytosis is higher on day 1, lower on days 3 and 7, the intensity of neutrophil phagocytosis is higher on days 1, 3 and 7, the activity of spontaneous and intensity of induced The HCT test is lower on days 1 and 30, the intensity of the spontaneous HCT test is lower on days 3, the activity of the induced HCT test is lower on days 1, higher on days 3 and 7.
Discussion. Thus, an analysis of the cellular composition and functional activity of innate immune cells in AS revealed an increase in the number of neutrophils of various degrees of maturity in the blood: metamyelocytes, rod-shaped and segmented neutrophils, activation of the absorption capacity and oxygen-dependent metabolism of blood neutrophils with a limitation of their functional reserve due to a decrease in the parameters of the induced HCT test. These changes reflect the involvement of neutrophils as the main cells of the innate immune system in anti-infective protection in AS and the likelihood of developing secondary infectious, hospital-acquired complications, and systemic inflammatory response syndrome [1, 12, 3]. 
In patients with a favorable prognosis of AS, the number of neutrophil subpopulations in the blood is lower than in patients with an unfavorable prognosis. In addition, with an unfavorable outcome of AS, the number of monocytes in the blood increases. It has been shown that in AS, some of the mature neutrophils die by apoptosis, and stimulation of the bone marrow myeloid outgrowth leads to an increase in the number of immature neutrophils in the blood [7]. Hyperactivation of neutrophils by proinflammatory mediators in AS leads to the release of large amounts of cytokines, defensins, and reactive oxygen species (ROS), which leads to damage to host tissues and the development of multiple organ failure [9, 13]. The pathogenic role in AS is played not by the total number of neutrophils, but by the presence of their hyperactive subpopulation involved in tissue damage [8]. In this regard, immature neutrophils (metamyelocytes, rod-shaped forms) are of particular importance, which have increased adhesive properties and easily penetrate into tissues, where they release reactive oxygen species [4, 9]. 
   In the group of patients with an unfavorable outcome of AS, there was a high activity and intensity of phagocytosis, as well as the activity and intensity of the spontaneous HCT test, which is associated with increased oxidative metabolism in activated phagocytes (respiratory explosion) with the formation of primary (superoxide anion, hydrogen peroxide, hydroxyl radical, singlet oxygen) and secondary (hypochloric acid, chloramine, products of lipid peroxidation) of metabolites with potent bactericidal activity. By day 7, the activity and intensity of the spontaneous HCT test are restored, and the functional reserve of neutrophils in terms of the intensity of the induced HCT test decreases. These changes reflect defects in oxygen-dependent mechanisms of microbicidal activity and can serve as a prognostic criterion for an unfavorable outcome in AS. 
Conclusions. 
1. It was found that in patients with AS, over the course of 1-7 days of follow-up, regardless of the outcome, the number of metamyelocytes, rod-shaped, segmented neutrophils increases in the blood; in comparative terms, with an unfavorable outcome of AS, neutrophilic leukocytosis is more pronounced on 1-7 days of follow-up, the number of monocytes increases on the 3rd day of follow-up.
2. In patients with AS, regardless of the outcome, the absorption capacity of blood neutrophils increases, oxygen-dependent metabolism in a spontaneous mode, oxygen-dependent metabolism in an induced mode is inhibited; in comparative terms, with an unfavorable outcome of AS, there is an escalation in the growth of the absorption capacity of blood neutrophils on days 1-7 of follow-up against the background of inhibition of oxygen-dependent neutrophil metabolism in a spontaneous mode. and induced modes mainly on the 3rd and 7th days of observation.

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About the authors

Michael Osikov

Federal State Budgetary Educational Institution of Higher Education "South-Ural State Medical University" of the Ministry of Healthcare of the Russian Federation, Chelyabinsk, Russia;
Chelyabinsk Regional Clinical Hospital State Budgetary Healthcare Institution, Chelyabinsk, Russia

Email: ritkaboyko@yandex.ru
ORCID iD: 0000-0001-6487-9083
Scopus Author ID: 16040195100

Doctor of Medical Sciences, Professor, Head of the Department of Pathophysiology, Head of the Department for Scientific and Innovative Work

Russian Federation

Margarita Boyko

Federal State Budgetary Educational Institution of Higher Education "South-Ural State Medical University" of the Ministry of Healthcare of the Russian Federation, Chelyabinsk, Russia

Email: ritkaboyko@yandex.ru
ORCID iD: 0000-0002-4046-2424
SPIN-code: 2646-0335
Scopus Author ID: 57218667153

PhD, Associate Professor of the Department of Pathophysiology

Russian Federation

Alexey Konashov

Federal State Budgetary Educational Institution of Higher Education "South-Ural State Medical University" of the Ministry of Healthcare of the Russian Federation, Chelyabinsk, Russia;
State Autonomous Healthcare Institution of the Order of the Badge of Honor "City Clinical Hospital No. 8, Chelyabinsk, Gorky str., 28, Chelyabinsk, Russia

Email: ritkaboyko@yandex.ru

Candidate of Medical Sciences, Associate Professor of the Department of Pathophysiology

Russian Federation

Andrey Gusev

Federal State Budgetary Educational Institution of Higher Education "South-Ural State Medical University" of the Ministry of Healthcare of the Russian Federation, Chelyabinsk, Russia;
Chelyabinsk Regional Clinical Hospital State Budgetary Healthcare Institution, Chelyabinsk, Russia

Email: ritkaboyko@yandex.ru

Assistant Professor of the Department of Pathophysiology, Federal State Budgetary Institution of Higher Education "South Ural State Medical University" of the Ministry of Health of the Russian Federation, anesthesiologist-intensive care unit "Chelyabinsk Regional Clinical Hospital" of the Ministry of Health of the Chelyabinsk Region, Chelyabinsk

Russian Federation

Vladislav Konashov

Federal State Budgetary Educational Institution of Higher Education "South-Ural State Medical University" of the Ministry of Healthcare of the Russian Federation, Chelyabinsk, Russia;
State Autonomous Healthcare Institution of the Order of the Badge of Honor "City Clinical Hospital No. 8, Chelyabinsk, Gorky str., 28, Chelyabinsk, Russia

Author for correspondence.
Email: ritkaboyko@yandex.ru

Assistant Professor at the Department of Pathophysiology, Federal State Budgetary Institution of Higher Education "South Ural State Medical University", trainee physician at the Intensive Care Unit of the State Medical Institution "City Clinical Hospital No. 8", Chelyabinsk

Russian Federation

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