Changes in the cytokine profile in patients with chronic rhinosinusitis of different phenotypes

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Introduction
Among all inflammatory diseases of the ENT organs, chronic rhinosinusitis occupies a leading position in prevalence in the world. In Russia, approximately 1 million 500 thousand people suffer from chronic rhinosinusitis, and in the USA this figure reaches 30-35 million people or 4.9 per 10,000 population [1]. Numerous studies show that the pathogenesis of this disease is influenced by the development of both a chronic inflammatory process in the mucous membrane of the nasal cavity and paranasal sinuses and allergic mechanisms [2]. Today, it is believed that three theses can be formulated on the heterogeneous nature of chronic rhinosinusitis: the inflammatory process in CRS is triggered by a dysfunctional interaction between exogenous agents inhaled through the nose and the human immune system; the importance of specific causative factors varies in individual patients, which leads to the formation of different types of tissue inflammation (endotypes); The clinical characteristics (phenotypes) of the disease and the response to treatment will depend on the first two points [3, 4]. Currently, chronic rhinosinusitis is divided into several endotypes depending on the predominant type of inflammation: T2 inflammation, non-T2 inflammation, and epithelial endotype. Each type of inflammation is characterized by the composition of both activated cellular elements and cytokines involved in it, which are produced by both effector and regulatory cells, including subpopulations such as T-helpers-17 (Th17) and T-helpers-22 (Th22). However, due to the overlap of biological functions of many molecules and the structural features of their receptors, the classification of cytokines is a complex task. According to the structural and functional classification of cytokines proposed by A.S. Simbirtsev, 13 families of cytokines can be distinguished, which differ in their main biological functions: interferons: INF-α, INF-β, INF-γ, etc. (antiviral activity, antiproliferative, immunomodulatory action); superfamily of interleukin type 1 and fibroblast growth factor: IL-1α, IL-1β, IL-18, IL-33, etc. (pro-inflammatory action, activation of specific immunity); chemokines: CC, CXC (IL-8), CX3C, C (regulation of chemotaxis of various types of leukocytes); cytokines of T-helper clones and regulatory functions of lymphocytes: T-helpers type 1 (IL-2, IL-15, IL-21, INF-γ, TNF) (activation of mononuclear inflammation and cellular immunity), T-helpers type 2 (IL-4, IL-5, IL-10, IL-13) (activation of allergic inflammation and humoral immunity), T-helpers type 17 (IL-17A, IL-22) (activation of neutrophilic inflammation and immunity against fungi), etc. [5].

The development and progression of the pathological process leads to a change in the level of individual cytokines necessary for the formation of an inflammatory response, thereby ensuring the process of blocking and eliminating the pathogen from the body. Cytokines are key factors that trigger and modulate intercellular communication in immunological reactions, the development of hematopoiesis, as well as host responses to infectious agents and inflammatory triggers. Thus, cytokines play a significant role in the pathogenesis of CRS, especially in maintaining the inflammatory response and attracting eosinophils. At the same time, the significance of various cytokines in the pathogenesis of CRS has not been definitively determined.
The aim of this study was to investigate cytokine regulation in patients with chronic rhinosinusitis depending on the phenotype of the disease.

Materials and methods
The study was conducted at the Clinical Pathophysiology Laboratory of the Federal State Budgetary Scientific Institution "Federal Research Center "Krasnoyarsk Scientific Center of the Siberian Branch of the Russian Academy of Sciences", a separate division of the Research Institute for Medical Problems of the North. The object of the study were patients with chronic rhinosinusitis, hospitalized in the ENT department of the institute's clinic, and selected using the continuous sampling method. As part of the clinical examination of patients, an integrated approach was used, which covered not only the collection of information on complaints and medical history, but also objective examinations, including specialized checks of ENT organs. Additionally, visualization methods such as X-ray and computed tomography of the sinuses were used in accordance with the latest medical standards of 2022. An important part of the diagnosis was also taking into account all the patient's acute and chronic diseases during the examination by an otolaryngologist. The diagnoses were coded according to the statistical classification of diseases, injuries and causes of death (ICD-10).
The main criteria for including patients in the study were the presence of complaints of frequent, prolonged discharge from the nasal cavity during the year, accompanied by pain, discomfort, a feeling of pressure or distension in the paranasal sinuses, the diagnosis was verified according to the clinical recommendations of the Ministry of Health (2022). Patients with infectious and acute diseases of other organs and systems, exacerbation and decompensation of chronic concomitant somatic diseases and those who refused to take part in the study were excluded from the study.
A total of 61 patients with chronic rhinosinusitis (48.7±3.9 years) were selected, of which 13 patients (43.7±3.3 years) had chronic allergic rhinosinusitis (CRS), 10 patients (45.7±4.3 years) had chronic polypous rhinosinusitis, 24 patients (44.7±3.3 years) had chronic infectious rhinosinusitis, and 14 patients (47.7±4.2 years) had chronic hyperplastic rhinosinusitis. The control group consisted of 30 practically healthy blood donors (46.7±3.5 years) comparable in gender and age with the study groups, undergoing routine examination at the institute's clinics. The levels of TNF-α, TNF-β, IFNγ, TGF-β, IL-1β, IL-2, IL-6, IL-8, IL-17A, IL-18, IL-10 and IL-4 in the blood serum of patients and healthy individuals were determined by the ELISA method on a Thermo Scientific Multiskan FC ELISA analyzer (Thermo Fisher Scientific, USA) using reagent kits manufactured by Vector-Best CJSC (Novosibirsk). Based on the results of the study, a database was formed on a personal computer in the MSExcel 2010 spreadsheet package. Statistical data processing was performed using the Statistica for Windows 8.0 (StatSoftInc., USA, 2008) and Microsoft Excel, 2007 (Microsoft, USA) application software packages. Processing of the obtained data included calculating nonparametric data: median (Me) and percentiles (C25-C75). The statistical significance of differences was determined using the Mann–Whitney rank test. The critical level of significance when testing statistical hypotheses was taken to be p <0.05.

Results and discussions
In all patient groups, there was a decrease in TNF-α compared to the control group (p1-2<0.001; p1-3=0.02; p1-4=0.02; p1-5<0.001). When studying TNF-β, a decrease in this indicator was found in all patient groups compared to the control (p1-2=0.03; p1-3=0.04; p1-4=0.02; p1-5=0.02). In patients with infectious chronic rhinosinusitis, there was a decrease in IFNγ compared to the control group (p1-4=0.04). When studying IL-1β, a decrease in this indicator was found in all study groups compared to the control (p1-2=0.02; p1-3<0.001; p1-4=0.01; p1-5<0.001). All patients with chronic rhinosinusitis had an increase in IL-2 levels compared to the control group (p1-2<0.001; p1-3<0.001; p1-4<0.001; p1-5<0.001). When studying IL-8, a decrease in this indicator was found in all studied patients compared to the control group (p1-2=0.001; p1-3=0.001; p1-4=0.002; p1-5=0.001). In all studied groups of patients, there was a decrease in IL-17A compared to the control group (p1-2<0.001; p1-3<0.001; p1-4<0.001; p1-5<0.001). When studying IL-18, it was revealed that in all groups there was a decrease in this cytokine compared to the control group (p1-2<0.001; p1-3<0.001; p1-4<0.001; p1-5<0.001). In all patients with chronic rhinosinusitis, there was a decrease in IL-4 compared to the control group (p1-2<0.001; p1-3<0.001; p1-4<0.001; p1-5<0.001). Conclusions
When studying the nature of cytokine regulation of various phenotypes of CRS, the presence of a Th1 variant of the immune response was found: in patients with chronic allergic and chronic polypous rhinosinusitis, there was a decrease in TNF-α, TNF-β, IL-1β, IL-8, IL-17A and IL-4, as well as an increase in IL-2 and IL-18. In patients with chronic infectious rhinosinusitis, the following was revealed: a decrease in TNF-α, TNF-β, IFNγ, IL-1β, IL-8, IL-17A and IL-4, as well as an increase in IL-2 and IL-18. In the group with chronic hyperplastic rhinosinusitis, there was a decrease in: TNF-α, TNF-β, IL-1β, IL-8, IL-17A and IL-4, as well as an increase in IL-2 and IL-18.

About the authors

Olga Valentinovna Smirnova

Federal Research Center "Krasnoyarsk Scientific Center of the Siberian Branch of the Russian Academy of Sciences" of the separate division of the Research Institute of Medical Problems of the North. Head of the Department of Medical Biology of the Siberian Federal University

Author for correspondence.
Email: ovsmirnova71@mail.ru
ORCID iD: 0000-0003-3992-9207

Doctor of Medical Sciences, Professor, Head of the Laboratory of Clinical Pathophysiology

Russian Federation, 660062, Krasnoyarsk, Partizana Zheleznyaka st., 3G

A A Sinyakov

Krasnoyarsk Science Centre of the Siberian Branch of Russian Academy of Science (FRC KSC SB RAS) separate division “Scientific Research Institute of medical problems of the North” (SRI MPN)

Email: sinyakov.alekzandr@mail.ru
ORCID iD: 0000-0002-4474-1893

Senior Researcher, PhD in Biology

Russian Federation, 660062, Krasnoyarsk city, Partisan Zeleznyaka str., 3G

References

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