Functional Activity Of Neutrophilic Granulocytes Research Articles

Disruption of the Functional Activity of Neutrophil Granulocytes as a Risk Factor for the Development of Lung Damage in Pregnant Women with COVID-19.

Currently, the assessment of immune status in patients with COVID-19 is limited to determining the count of polymorphonuclear leukocytes and the phagocytic function of neutrophils, which is insufficient to understand the regulatory role of innate immunity cells in the development of pneumonia. However, no such studies have been conducted in pregnant women with COVID-19. The aim of this study was to investigate the functional state of neutrophil granulocytes in order to identify predictors of pneumonia severity risk in pregnant women with COVID-19. A clinical characterization of pregnant women with COVID-19 in addition to minimal and average lung changes was provided. The composition and ratio of morphological forms of leukocyte cells were studied. Cytochemical studies of neutrophil granulocytes were carried out and calculations of the mean cytological index (MCI) for succinate dehydrogenase, myeloperoxidase, and cationic proteins were performed. The number of NETs in blood smears was counted. Independent predictors of pneumonia severity in pregnant women with COVID-19 were calculated using regression analysis. The quality of the model was assessed using ROC analysis. In pregnant women with COVID-19 and an average volume of lung changes, the number of polymorphonuclear leukocytes (p = 0.03) and band neutrophils (p = 0.002) in the blood was significantly higher than in pregnant women with minimal lung changes. The MCI indicators of succinate dehydrogenase, cationic proteins, and myeloperoxidase in pregnant women with COVID-19 were reduced in relation to the control group (p < 0.0001). In blood smears of pregnant women with COVID-19 and an average volume of lung changes, the number of NETs increased (p = 0.002). Regression analysis showed that succinate dehydrogenase and NETs are independent predictors of pneumonia severity in pregnant women with COVID-19. Our study confirms the prognostic significance of low levels of neutrophilic succinate dehydrogenase and high levels of NETs in the blood of pregnant women with COVID-19. The combination of these two biomarkers is a significant reflection of the severity of pneumonia development in pregnant women with COVID-19. However, further research is needed to identify the mechanisms underlying this association.

Positive effects of recombinant interferon α2b on the phenotype of CD16&lt;sup&gt;+&lt;/sup&gt;INFα/βR1&lt;sup&gt;-&lt;/sup&gt;CD119&lt;sup&gt;+&lt;/sup&gt;, CD16&lt;sup&gt;+&lt;/sup&gt;INFα/βR1&lt;sup&gt;+&lt;/sup&gt;CD119&lt;sup&gt;-&lt;/sup&gt; neutrophil granulocyte subset in patients with post-COVID syndrome and herpesvirus infections

Post-COVID syndrome (PCS) is a multisystem inflammatory condition with manifestations of chronic fatigue syndrome (CFS) and cognitive disorders (CD), along with reactivation of chronic herpesvirus infections (HVI). The PCS manifestations require studying the molecular mechanisms associated with the production of IFN and receptor functions of neutrophil granulocytes (NG), which is relevant and promotes the search for immunotherapeutic strategies in patients with PCS. Our objective was to study the in vitro effects of recombinant interferon α2b (recIFNα2b) on the phenotype of CD16+IFNα/βR1-CD119+, CD16+IFNα/ βR1+CD119+ subsets and functional activity of NG in patients with post-COVID syndrome and herpesvirus infections. Materials and methods: 45 patients (24-60 years old) with PCS and HVI (HSV 1, EBV, HHV6, CMV) comprised the study group 1 (SG1). A questionnaire was conducted to assess the severity of PCS symptoms using a point scale. We performed a study of the content and phenotype of NG subsets, i.e., the CD16+IFNα/βR1-CD119+, CD16+IFNα/βR1+CD119-, CD16+IFNα/βR1+CD119+ subpopulation, phagocytic and NADPH oxidase function of NG before and after in vitro incubation with recIFNα2b (50 IU/ µL, for 60 min, at 37 °C) in the study group 1a (SG1a). The comparison group (CG) of 30 volunteers examined during the pre-COVID period. Results: We revealed more pronounced clinical manifestations of CFS and CD in SG1 patients with mixed HVI, than in mono-HVI cases. Increased expression density of all receptors was registered on CD16+IFNα/βR1+CD119-NG and CD16+IFNα/βR1-CD119+ NG, thus suggesting the NG activation with initiation of cytotoxicity or NETosis, a decrease in phagocytic function and intensity of NADPH oxidase activity with depletion of NG reserve capacity in SG1. We have obtained some data on the positive effect of recIFNα2b in vitro (SG1a), e.g., decreased CD16 expression density and enhancement of IFNα/βR1 receptor expression in the CD16+IFNα/βR1+CD119- subset. In the CD16+IFNα/βR1-CD119+ subset, we have found persistence of increased MFI CD16 and MFI CD119 receptors, restoration of defective NG phagocytic function and reduced excessive activity of NADPH oxidases. Conclusion: The positive effects of the recIFNα2b influence on deficient function of NG in PCS patients suggest an oppoptunity of using immunotherapy with a recIFNα2b-based drug, combined with highly active antioxidants for treatment of various PCS manifestations including CFS, CD, HVI, thus, probably, ensuring adequate functioning of antiviral and regulatory mechanisms of the immune system.

Local immune state in the women with miscarriage

Functional state of immunity provides maintenance of immunological tolerance for allogeneic fetus, and high level of local protection against antigenic stimulation. Increased functional activity of phagocytic cells at systemic and local levels may cause impairment of complete fetoplacental complex, thus leading to abortion. The purpose of our study was to assess the state of local immunity in women with pregnancy loss.&#x0D; The study involved 174 women in their reproductive age. The first group consisted of 65 patients (37.4%) with a diagnosis of miscarriage, the history of 2 abortions over the period from conception to the 20th week of gestation. The second group included 37 patients (21.2%) with documented recurrent pregnancy loss, with a history of 3 miscarriages. The control group consisted of 72 conditionally healthy women (41.4%) who had 2 or more term pregnancies with the same partner, without a history of obstetric and gynecological complications. Their mean age was 366 years old. Total number and viability of leukocytes, the indices of functional neutrophil activity, their functional reserve and neutrophil stimulation index were determined in cervical mucosal samples over the first phase of the menstrual cycle.&#x0D; In the first phase of menstrual cycle, the women with miscarriage and recurrent pregnancy loss exhibited a statistically significant increase of phagocytosis by the neutrophils from cervical mucus as compared to the group of conditionally healthy women. Intensity of neutrophil phagocytosis in cervical mucus reached higher values in the women with recurrent pregnancy loss more often, compared to the control group. Evaluation of the functional reserve of cervical mucus neutrophils in the subjects with recurrent pregnancy loss and miscarriage showed a trend towards statistically significant differences: this parameter was higher in the patients than in control group. The ability of cervical mucosal neutrophils to produce reactive oxygen species (both spontaneous and induced) did not show statistically significant differences between the patients and controls.&#x0D; Hence, we have observed aberrant functional activity of neutrophilic granulocytes from cervical mucosa in the groups of women with 2 or more abortions, without changing ability of the cells to produce reactive oxygen species. This finding may be explained by prevalence of oxygen-independent mechanisms of intracellular killing, thus suggesting a role of neutrophils for impaired balance of immunological tolerance in pregnant women at the local level.

Phagocyte Activity in Response to Exposure to Methicillin-Resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt; Strains

The research focuses on the study of phagocyte activity of blood neutrophilic granulocytes, monocytes, and eosinophils under the exposure to methicillin-resistant strains of Staphylococcus aureus bacteria. The subjects of the research were represented by blood neutrophilic granulocytes, monocytes, and eosinophils, isolated from healthy people, as well as methicillin-resistant and methicillin-sensitive S.aureus bacteria strains (MRSA and MSSA). Phagocytosis functions (phagocytic number and phagocytic index) were estimated by FITC-labeled bacteria. The analysis of stained cells was performed using FC-500 flow cytofluorimeter (Beckman Coulter, USA) for whole peripheral blood. Phagocytosis research resulted in the following findings. In response to MRSA, the percentage of neutrophils involved in phagocytosis and average number of bacteria being present inside the cells increase as compared to those within sensitive strains. In response to MRSA exposure, the phagocytic index is higher in monocytes with CD14+CD16¯phenotype. At the same time, phagocytic number is higher in non-classic populations of monocytes CD14+CD16+and CD14lowCD16+. Moreover, it should be noted that the classical type of monocytes CD14+CD16¯tends to be activated faster in regard to MRSA, but phagocytosis efficiency is lowered. Blood eosinophils also actively respond to MRSA. Thus, changes in functional activity of neutrophilic granulocytes, monocytes, and eosinophils in peripheral blood were established during MRSA induction. The bacterial receptor apparatus changes due to the modification of cellular wall as a result of acquired resistance to antibiotics.

Functional activity of blood neutrophilic granulocytes in patients with opisthorchiasis, depending on the severity of liver fibrosis

To study the activity of neutrophilic granulocytes in patients with opisthorchiasis, depending on the severity of liver fibrosis. A total of 74 patients with chronic opisthorchiasis (39 men and 35 women, average age 42.3 years) and 32 practically healthy patients (17 men and 15 women, average age 41.5 years) aged 24 to 60 years were examined. Diagnosis of opisthorchiasis was carried out by two methods: coprooscopy and identification of eggs or bodies of adult parasites in duodenal content. Liver fibrosis was determined by the method of elastometry according to the METAVIR scale in all 74 patients with opisthorchiasis. The study of the functional activity of neutrophils in the blood was performed to all 74 patients with opisthorchiasis and 32 healthy individuals from the control group by chemiluminescent analysis with measurement of the reactive oxygen species (ROS) production intensity in a spontaneous and zymosan-induced reaction in lucigenin and luminol-dependent processes. Liver fibrosis F2 by METAVIR was registered in 20.3% of the examined individuals, liver fibrosis F3F4 by METAVIR was detected in 17.6% of patients with opisthorchiasis. In patients with opisthorchiasis with liver fibrosis F3F4 by METAVIR, a significant decrease in the functional activity of neutrophilic granulocytes was registered in comparison with individuals with liver fibrosis F0F1 by METAVIR, as evidenced by a significant decrease in the maximum intensity of ROS production (Imax) and the area under the curve (S) chemiluminescence in lucigenin and luminol-dependent processes both in the spontaneous and zymosan-induced reaction. These results provide new information to explain the mechanisms of liver fibrosis in patients opisthorchiasis and create opportunities for the development of diagnostics and preventive technologies.

Особливості імунної відповіді організму піддослідних тварин на дію мийних засобів на основі аніонних поверхнево-активних речовин, призначених для догляду за дитячим посудом

Objective: We assessed the impact of the detergents, based on anionic surfactants, for the care of the children’s tableware on the indicators of the immune system of the experimental animals. Materials and methods: The effect of aqueous solutions of Alionka, Karapuz, Ushastyi Nian, Frosch Baby detergents on the indicators of the immune system was performed in the experiment with guinea pigs during 21 days. We determined the number of T- and B-lymphocytes, the number of neutrophilic granulocytes and active phagocytes; studied the reaction of the precipitation of circulating immune complexes with a solution of polyethylene glycol 6000. We used the methods of statistical processing of the results of medico-biological research. Results: The obtained results showed that the epicutaneous impact of all studied detergents for the care of children’s tableware could cause the immunomodulatory effects in the organism of guinea pigs. Changes in the largest number of the indicators of the immune system, which affected all links, were observed when exposed to Ushastyi Nian detergent. The least pronounced changes of the immunity were observed when exposed to the Frosch Baby detergent and they concerned only the indices of nonspecific resistance of the organism. The revealed inhibition of the functional activity of neutrophilic granulocytes, which was observed in animals of all experimental groups, may indicate a decrease in the protective functions of the organism of guinea pigs.

IN VITRO EXPERIMENTAL REWIRING OF 4 NEUTROPHILIC GRANULOCYTE SUBSETS FROM THE PRO-INFLAMMATORY TO THE ANTI-INFLAMMATORY PHENOTYPE IN CHILDREN WITH SURGICAL PURULENT INFECTION OF SOFT TISSUE

Treatment of young children with atypical or recurrent purulent soft tissue infections (PSTD) that do not respond well to surgery and antibiotics is most challenging. PSTD occurs against the background of impaired functioning of the immune system and, first of all, the system of neutrophilic granulocytes (NG). The vector effect of immunotropic therapy on a specific NG subsets may allow the correction of NG dysfunctions without compromising host protection, including strategies to enhance, inhibit or restore their functions.The aim of study: to evaluate in vitro the modulating effects of arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine (HP) on the transformed phenotype of 4 NG subsets, as well as on the functional activity of NG in children with purulent-inflammatory soft tissue diseases.We studied samples of peripheral blood (PB) from young children 2-4 years old: 17 children with atypical acute PSTD and 10 apparently healthy children. At stage I, a comparative assessment of the content and phenotype of 4 NG subsets CD16+CD62L+СD63- , CD16+CD62L+СD63+, СD64- CD16+CD32+CD11b+, СD64+CD16+CD32+CD11b+, phagocytic and microbicidal functions of NG was carried out. At stage II, the in vitro system determined the effects of HP on NG in children with PSTD according to the studied parameters. By the method of flow cytometry (FC500 “Beckman Coulter” (USA), conjugates of MkAT “Beckman Coulter International S.A.” (France)), the relative number of NGs of the studied subsets and the density of receptor expression (MFI) were determined. To assess the phagocytic function of NG a microbiological method was used to assess the completeness of phagocytosis with S. aureus (strain 209). The activity of NG NADPH oxidase was investigated in the NBT-spontaneous test (NBTsp.) and in the in vitro NBT-induced test (NBTind.). A comparative study of PB samples from conventionally healthy children and children with PSTD made it possible to identify various variants of transformation of the phenotype of the studied NG subsets, associated with defects in their functional activity. In the in vitro system the effects of HP were demonstrated, manifested by a decrease in the amount of CD16+CD62L+CD63+NG and an increase in CD16+CD62L+CD63- NG, modulation of the negatively altered phenotype of subsets CD64- CD32+CD16+CD11b+NG and CD64+CD32+CD16+CD11b+NG, aimed at restoring phagocytic function and maintaining the tension of NADPH oxidases.As a result of the study it was found the immunomodulatory effects of HP, which is manifested in the reorientation of NG from the pro-inflammatory phenotype to the anti-inflammatory one, which can be used in the future when creating personalized targeted immunotherapy aimed at correcting defective functioning NG in early children, suffering from PSTD.

IMMUNOLOGICAL CRITERIA FOR EFFECTIVE SYSTEMIC USE OF MURAMYL PEPTIDE IMMUNOMODULATOR IN THE TREATMENT OF PATIENTS WITH BURN INJURIES

Abstract. Introduction. The problem of providing effective care to patients with burn injuries remains far from being finally solved. Due to significant immune disorders that occur against the background of thermal damage, the use of immunomodulatory drugs remains promising. Therefore, the aim of this study was to analyze the effectiveness of systemic use of the immunomodulatory drug Liastenum® in the complex treatment of patients with burn injuries.&#x0D; Materials and methods: The study involved 35 patients with burn injuries with the index of damage severity ranging from 30 to 60 units. Patients in the main group (n = 15) in addition to the usual treatment received an immunomodulator Liastenum®, which was administered intramuscularly at a dose of 2 mg once every 3 days (course dose — 10 mg). Treatment of patients in the comparison group (n = 20) did not involve the use of drugs with targeted immunocorrective action. Examination of patients included laboratory immunological examination of venous blood with assessment of various parts of the immune system and determination of the “Neutrophilic granulocytes form factor” indicator, which was performed on the 3rd, 14th, 21st day after the injury.&#x0D; The results of the study and their discussion. The obtained results allowed to confirm the ability of Liastenum® to stimulate the functional activity of neutrophilic granulocytes, restoring the phagocytic reserves of these cells, to normalize the level of lymphocytes and their individual subpopulation forms. In general, the action of the immunomodulatory drug was characterized by a balanced effect on the cellular, humoral parts of the immune system, phagocytosis.&#x0D; Conclusions. The effectiveness of systemic use of immunomodulatory drug Liastenum® in a comprehensive program of care for patients with burn injuries by balanced correction of immune disorders was laboratory established.

ДИФФЕРЕНЦИРОВАННАЯ ТРАНСФОРМАЦИЯ ФЕНОТИПА СУБПОПУЛЯЦИЙ НЕЙТРОФИЛЬНЫХ ГРАНУЛОЦИТОВ ПРИ ВИРУСНО-ВИРУСНЫХ И ГНОЙНЫХ БАКТЕРИАЛЬНЫХ ИНФЕКЦИЯХ У ИММУНОКОМПРОМЕТИРОВАННЫХ ДЕТЕЙ

Children with clinical signs of immunocompromise have impaired functioning of the immune system. At the same time, the state of subpopulations of multifunctional neutrophil granulocytes (NG), which provide antiviral and antibacterial protection, in such patients has been poorly studied. Objective of the study: to assess the features of the transformation of the phenotype of the three most significant subpopulations of NG, their association with impaired phagocytic and microbicidal activity in immunocompromised children with atypical viral-viral co-infections and bacterial infections, and to clarify their differential diagnostic significance. Materials and methods of research: the phenotypes of NG subpopulations expressing CD64, CD32, CD16, CD11b receptors, phagocytic and microbicidal activity of NG were studied in the peripheral blood of 43 immunocompromised children of both sexes aged of 5–9 years old suffering from recurrent ARVI, atypical chronic mono – or mixed herpes virus infections (HVI) and purulent bacterial infection. 3 study groups were formed: 1st – repeated ARVI/HVI mono, 2nd – repeated ARVI/HVI mixed, 3rd – minor purulent infection (MPI) and a comparison group (20 apparently healthy children). Results: different differentiated transformation of the phenotype of functionally significant subpopulations of NG associated with impaired functional activity of NG as well as with the incidence of viral co-infections (repeated ARVI and recurrent HVI mixed) and the severity of clinical manifestations of MPI has been identified in the studied groups. Conclusion. Еvaluation of the features of transformation of phenotype of NG subpopulations, their effector functions in immunocompromised children with atypical viral-viral co-infections and purulent bacterial infections, will allow in the future to optimize the methods of target immunotherapy aimed at remodulating the negatively transformed phenotype of NG subpopulations, correcting their dysfunctions and, thereby, significantly increase the clinical effectiveness of therapeutic and preventive measures.

Biogenic Ferrihydrite Nanoparticles: Synthesis, Properties In Vitro and In Vivo Testing and the Concentration Effect.

Biogenic ferrihydrite nanoparticles were synthesized as a result of the cultivation of Klebsiella oxytoca microorganisms. The distribution of nanoparticles in the body of laboratory animals and the physical properties of the nanoparticles were studied. The synthesized ferrihydrite nanoparticles are superparamagnetic at room temperature, and the characteristic blocking temperature is 23–25 K. The uncompensated moment of ferrihydrite particles was determined to be approximately 200 Bohr magnetons. In vitro testing of different concentrations of ferrihydrite nanoparticles for the functional activity of neutrophilic granulocytes by the chemiluminescence method showed an increase in the release of primary oxygen radicals by blood phagocytes when exposed to a minimum concentration and a decrease in secondary radicals when exposed to a maximum concentration. In vivo testing of ferrihydrite nanoparticles on Wister rats showed that a suspension of ferrihydrite nanoparticles has chronic toxicity, since it causes morphological changes in organs, mainly in the spleen, which are characterized by the accumulation of hemosiderin nanoparticles (stained blue according to Perls). Ferrihydrite can also directly or indirectly stimulate the proliferation and intracellular regeneration of hepatocytes. The partial detection of Perls-positive cells in the liver and kidneys can be explained by the rapid elimination from organs and the high dispersion of the nanomaterial. Thus, it is necessary to carry out studies of these processes at the systemic level, since the introduction of nanoparticles into the body is characterized by adaptive-proliferative processes, accompanied by the development of cell dystrophy and tension of the phagocytic system.

Metabolic activity of peripheral blood neutrophils in patients with &lt;i&gt;Opisthorchis felineus&lt;/i&gt; invasion and skin syndrome

The aim was to examine functional activity of peripheral blood neutrophilic granulocytes in patients with Opisthorchis felineus invasion and skin syndrome. Materials and methods. A total of 92 patients with chronic opisthorchiasis were examined, of which 38 patients (20 males and 18 females, average age 35.7±3.9 years) and 54 patients (28 males and 26 females, average age 36.5±4.1 years) with/without skin syndrome, as well as 32 apparently healthy subjects (17 males and 15 females, average age 41.5 years). Detection of adult parasite eggs or bodies in the duodenal contents and/or feces was used as the major diagnostic method of opisthorchiasis used in all 92 opisthorchiasis patients. Liver fibrosis was assessed by elastometry method according to the METAVIR scale in all 92 opisthorchiasis patients. Functional activity of peripheral blood neutrophilic granulocytes was examined in all 92 opisthorchiasis patients and 32 healthy individuals from the control group by chemiluminescent analysis measuring intensity of reactive oxygen species production in spontaneous and zymosan-induced reactions in lucigenin and luminol-dependent processes. Results. Opisthorchiasis patients with vs. without skin syndrome suffered at higher rate from pain in the right hypochondrium, dyspepsia, astheno-vegetative syndrome, skin itching, blood eosinophilia, hyperbilirubinemia, cholestatic syndrome, cholecystitis, hepatomegaly, and had elevated serum immunoglobulin E level. Liver fibrosis F3—F4 according to METAVIR was determined in 18.4% and 11.1% of opisthorchiasis patients with vs. without skin syndrome (OR = 1.78; CI 0.57—5.57; p = 0.05). Both luminol- and lucigenin-dependent processes in opisthorchiasis patients with vs. without skin syndrome revealed significantly decreased functional activity of neutrophilic granulocytes which was evidenced by significantly decreased maximum intensity of reactive oxygen species production and the area under the chemiluminescence curve both in spontaneous and zymosan-induced reaction. Conclusion. The results suggest that the presence of skin syndrome in patients with opisthorchiasis is accompanied by more prominent clinical and laboratory manifestations, decreased functional activity of peripheral blood neutrophils, thereby allowing to consider it as a marker of intense pathology.

Chemiluminescence activity of neutrophil granulocytes under the influence of magnetic nanoparticles of ferrihydrite (&lt;i&gt;in vitro&lt;/i&gt;)

The article presents the results of studying and evaluating the impact of magnetic nanoparticles of ferrihydrite on neutrophil granulocytes in human blood ( in vitro ) in order to determine their bio compatibility and eco toxicity. The subject soft here search were blood neutrophil granulocytes of 29 conditionally-healthy donors of blood, as well as magnetic nanoparticles of ferrihydrite (NP), the preparation dose in minimum concentration reached 25 mg, in maximum concentration it reached 50 mg per 10 6 cells/ml. We implemented the sol of magnetic NP, obtained by biogenic synthesis in International Scientific Centre for Studying Extreme States of an Organism. Functional activity of blood neutrophil granulocytes has been determined by luminol-dependent chemiluminescence. Magnetic NP were introduced into pilot samples straight before chemiluminescent analysis, and also after the incubation with in 30 minutes under 37 °С entigrade. As a result of thee stimation of the early response of neutrophil granulocytes to the influence of minimum concentration of magnetic NP in vitro we found statistically true decrease of the intensity (1.6 times), the area under the curve (2.1 times) in zymosan-induced chemiluminescent response, the activation index (2.3 times). When evaluating the late response of neutrophil granulocytes to the influence of maximum concentration of magnetic NP in vitro we have found statistically true lowering of the time of reaching the peak (10 times) of spontaneous chemiluminescence. More over we marked consider able lowering of maximum intensity 6 times and the reduction of the area under the curve of zymosan-induced chemiluminescence 5.6 times under the influence of magnetic NP under the lowering of activation index 3.7 times. The authors determined that magnetic NP were intensively decreasing the functional activity of neutrophil granulocytes. The intensity of the impact is higher under the preliminary incubation of the cells with magnetic NP. At the same time, short effect of magnetic NP to neutrophil granulocytes can be a modulating one and depends on the initial level of cell reactivity. We revealed that magnetic NP influence concerns only activated cells.

Assessment of the functional activity of neutrophilic granulocytes in patients with metabolic syndrome

Assessment of the functional activity of neutrophilic granulocytes in patients with metabolic syndrome

Changes in Functional Activity of Neutrophils and Monocytes Isolated from the Peripheral Blood of Women at Different Phases of the Menstrual Cycle.

We studied functional activity of neutrophilic granulocytes and monocytes isolated from the peripheral blood of women during the follicular and luteal phases of the menstrual cycle. It was shown that phagocytic activity of neutrophilic granulocytes increases, their intracellular oxygen-dependent bactericidal activity decreases, and the number of monocyte extracellular traps increases in women in the luteal phase of the menstrual cycle in comparison with the follicular phase.

Peculiarities of the exposure of the antibiotics of the macrolid group on the functional activity of neutrophilic granulocytes of peripheral blood in vitro

Dose-dependent eff ects of antibiotics and their eff ect on immune cells of the body, in particular neu-trophilic granulocytes, are still poorly understood. The method of fl ow cytometry was determined by the minimum eff ective dose of chemomycin by assessing the degranulation of neutrophils. It was found that under the action of minimum eff ective dose chemomycin neutrophils form neutrophil extracellular traps (NETs), which are one of the mechanisms of adaptive immunity. It is noteworthy that the release of NETs in response to the action of chemomycin was not dependent on gender.

ВПЛИВ МАЗІ, ЩО МІСТИТЬ СУХИЙ ЕКСТРАКТ КОРЕНЯ СОЛОДКИ, НА ФАГОЦИТАРНУ АКТИВНІСТЬ ГРАНУЛОЦИТАРНИХ НЕЙТРОФІЛІВ КРОВІ В МУРЧАКІВ З ЕКСПЕРИМЕНТАЛЬНИМ ДЕРМАТИТОМ

У статті наведено результати вивчення впливу мазі «Алерголік», що містить сухий екстракт кореня солодки, на фагоцитарну активність гранулоцитарних нейтрофілів крові мурчаків з експериментальним алергічним контактним дерматитом, викликаним 2,4-динітрохлорбензолом. Згідно з отриманими даними, мазь «Алерголік», що застосовувалась в умовах лікувально-профілактичного режиму, мала виражену імунотропну дію, яка полягала в достовірному зниженні, відносно групи контрольної патології, фагоцитарної активності гранулоцитарних нейтрофілів, чого не спостерігали в групі тварин, яких лікували препаратом порівняння «Фладекс».

FEATURES OF FREE RADICAL OXIDATION AND ANTIOXIDANT PROTECTION IN CHILDREN WITH SUGAR DIABETES OF THE FIRST TYPE

Aim. Assessment of the state of lipid peroxidation, antioxidant defense and oxidative stress indicators on the model of functional activity of neutrophil granulocytes of blood in children with type 1 diabetes mellitus depending on the stage of endocrinopathy compensation. Materials and methods. The material of laboratory-diagnostic and clinical studies was the results of a survey of 38 children without endocrine pathology and 89 children with type 1 diabetes mellitus at various stages of disease compensation. The state of antioxidant protection was assessed taking into account the overall antioxidant activity, superoxide dismutase activity, retinol level, α-tocopherol, the content of reduced and oxidized glutathione. The intensity of lipoperoxidation processes was studied by the level of substrates with conjugated double bonds, diene conjugates, ketodienes, conjugated trienes, malonic dialdehyde, common lipids. Functional activity of neutrophil granulocytes was investigated by means of luminol-dependent chemiluminescence. Results. In children with type 1 diabetes mellitus, the intensification of lipid peroxidation processes and the activation of the antioxidant defense system, including variously directed changes in non-enzymatic mechanisms, are noted against the negative effect of active oxygen species. In children with decompensated type 1 diabetes mellitus, severe lipid metabolism disorders combined with oxidative stress indices increase the course of endocrine pathology, significantly increasing the likelihood of intravascular complications. Conclusion. The coordinated generation of active forms of oxygen and the processes of oxygen-dependent metabolism of blood cells in children with compensated type 1 diabetes mellitus indicates the development of the second stage (resistance) of oxidative stress. Reduction of reactive oxygen species production, reduced activation rate оxygendependent metabolism phagocytes incompleteness phagocytosis mechanisms, correlating with an increase in lesion area (destruction of insulin-producing β-cells) of the pancreas in children with decompensated type 1 diabetes, indicating the occurrence of the third step (debilitation) oxidative stress. Violations of metabolic pathways in children with type 1 diabetes are determined by the intensity of «respiratory burst» of neutrophil granulocytes in the «Lipid peroxidation – antioxidant protection».

HEMOSTASIS AND FUNCTIONAL PROPERTIES OF NEUTROPHILS IN PATIENTS WITH VARIOUS SENSITIVITY TO ACETYLSALICYLIC ACID IN ACUTE CORONARY SYNDROME

Aim. To study the specifics of relation between hemostasis parameters and functional activity of neutrophil granulocytes in sensitive and resistant to acetylsalicylic acid (ASA) patients with acute coronary syndrome. Material and methods. Totally 53 patients included during the first 24 hours of ACS. Controls consisted of 50 relatively healthy individuals. All patients before the treatment and revascularization were assessed for the resistance to ASA, and selected into groups of sensitive (AS) and resistant (AR). Sensitivity test was done in vitro. In all participants the coagulation and platelet clotting systems were studied. With the luminescent method the neutrophil activity was assessed. Results. In AR with ACS there was decrease of platelets in blood, decreased level of ADP- (ADP 5 mcM) and adrenalin-induced aggregation, increases the level of ADP-induced platelets aggregation (PA) with ADP 0,1 mcM, and von Willebrand factor level. In the ASA sensitive patients there was a decrease of adrenalin induced aggregation. There were no specifics of coagulation hemostasis according to ASA sensitivity. Chemiluminiscence (CL) of neutrophils in ACS has a maximized out time. In AR there was decrease of activation. In AS functional activity of neutrophils more significantly correlates with the parameters of clotting, but in AR — with fibrinolytic and anticoagulation parameters. Conclusion. In ACS patients depending on ASA sensitivity, there are character specifics of platelet clotting: increase of ADP-induced aggregation in AR, decrease of platelet amount and the level of von Willebrand factor. In ACS patients there is a decreased velocity of respiratory boost in neutrophils. In AR there is more prominent dysbalance of coagulation hemostasis and respiratory boost of neutrophils, with the decrease of neutrophil reactivity related to clotting activity and appearance of relations with the parameters of fibrinolytic and anticoagulation activity.

IMMUNOLOGICAL PARAMETERS IN PATIENTS WITH MONOINFECTION BY HUMAN PAPILLOMAVIRUS AND IN PATIENTS CO-INFECTED BY PAPILLOMAVIRUS AND UROGENITAL PATHOGENS

Aim of this study was to investigate the characteristics of cellular and humoral immunity and functional activity of neutrophilic granulocytes in women with monoinfection by human papillomavirus and combination of human papillomavirus (HPVI) and urogenital pathogens (UGI). It was established that more serious disorders in the immune system in women were revealed in case of mixed HPVI and UGI in compare with case of monoinfection by human papillomavirus. Only in patients with combined HPVI and UGI the content of total lymphocytes, T-cells and cytotoxic T lymphocytes was reduced. In patients with combined HPVI and UGI the humoral immunity was characterised by increased concentration of IgM, its level of relative synthesis and content of circulating immune complexes. Independently of the infection type in women the reduction of CD4+ lymphocytes, NK-cells, increased number of γδТ-lymphocytes, as well as increase the functional activity of blood neutrophils have been detected. However, in case of combined HPVI and UGI an inhibitory effect on the reserve capacities of phagocytic cells was revealed.

FEATURES OF LUMINOL- AND LUCIGENIN-INDUCED CHEMILUMINESCENCE OF NEUTROPHILIC GRANULOCYTES IN PATIENTS WITH CHRONIC RHINOSINUSITIS

Studies of functional activity of neutrophilic granulocytes from patients with chronic rhinosinusitis was performed using luminol- or lucigenin-dependent chemiluminescence. We have revealed increased production of reactive oxygen species (ROS) both under basal conditions, and in zymozan-induced chemiluminescent reaction, employing luminol as an activator, and determining total functional activity of neutrophils. Meanwhile, in lucigenin-induced chemiluminescence associated with production of superoxide anion radicals, a delayed ROS production was revealed, both in spontaneous and zymozan-induced response. One may suggest that, when studying these pathological events in neutrophils, ROS production in the cells proceeds mostly by myeloperoxidase pathway.