Phagocytic Activity Of Blood Neutrophils Research Articles

Phagocytic activity of neutrophil blood granulocytes in children with chronic non-specific lung diseases with post-inflammatory pneumofibrosis

One of the reasons for the development of chronic forms of lung diseases and pneumofibrosis as a pathological outcome of this inflammation may be an insufficient immune response. The key position belongs to neutrophilic granulocytes. Given the importance and renewed interest in neutrophils as tools for regulating immunity, leading to the progression of the fibrous process in children with CKD is an urgent problem. The purpose of the study was to study the phagocytic activity of blood neutrophils as an important component of innate immunity in patients with CNLD. Seventy children with CNLD with focal or segmental post-inflammatory pulmonary fibrosis were examined. Of these, 27 were children with progression of PPF (main group) and 43 children with PPF without progression (comparison group). The controls were the indicators of 23 healthy children, comparable by gender and age. The studies were carried out during clinical remission. The phagocytic activity of neutrophils (PНAN), phagocytic number (PN), spontaneous and stimulated NBT test were determined. The stimulation index of the NBT test (NBTst-NBTsp) was calculated. The mitochondrial membrane potential was determined using a BD FACS Calibur cytometer (USA) (BD Pharmigen, USA). PN in children with chronic heart disease with progressive PF was significantly reduced in comparison with patients in the comparison group and with the control group. The PHAN in children with CKD in both groups was within the standard values, however, it was lower than in the control group. The activity of the spontaneous HBT test in children of the main group was 1.7 times higher than in children in the comparison group and 3 times higher than in children from the control group. However, the stimulation index in the main group was significantly lower than in the control group. An increase in cells with a reduced membrane potential of granulocyte mitochondria was revealed in children of the main group compared with the control group. Children with CKD with the progression of fibrosis processes develop changes in the functional and metabolic activity of peripheral blood neutrophils typical for an inflammatory reaction, characterized by an increase in oxygen-dependent and a decrease in phagocytic cell activity.

Neuroendocrine-immune relationships at rats regardless of sex and exposure to stressors or adaptogens

Introduction and aim. Since the discovery of the famous Selye triad, studies of the connections between adaptation hormones and the immune system have remained relevant. We formulated the aim of the future study: in the mode of synchronicity, to reveal sexual differences in the parameters of the neuroendocrine-immune complex and the state of neuroendocrine-immune relationships in intact rats and exposed to stressors and adaptogens. This article initiates the beginning of the movement towards achieving the goal. Material and methods. The experiment is at 96 rats Wistar line: 48 males and 48 females. Over the 12 days, one male and female rat remained intact and 3 other pairs were exposed to chronic aversive stress for 6 days. We calculated the parameters of the HRV: Mode, Amplitude of the mode and Variational scope as markers of the circulating catecholamines, sympathetic and vagal tones respectively. Among endocrine parameters determined excretion of 17-Ketosteroides and serum levels of main adaptation hormones such as Corticosterone, Aldosterone, Testosterone, Triiodothyronine, as well as Parathyroid hormone and Calcitonin. The percentage of lymphocyte populations and the parameters of phagocytosis by neutrophils and monocytes of Staphylococcus aureus were determined in the blood. The Thymus and Spleen were weighed and made smears-imprints for counting Thymocytogram and Splenocytogram. Results. The canonical correlation between neuroendocrine and immune parameters was analyzed. The most pronounced immunomodulatory effect is exerted by catecholamines (R=0.928), sympathetic tone (R=0.849), and testosterone (R=0.829). Vagal tone (R=0.697), 17-ketosteroides (R=0.688), and aldosterone (R=0.686) moderately modulate immunity, while the immunomodulatory effects of triiodothyronine (R=0.610), corticosterone (R=0.584), PTH (R=0.510), and calcitonin (R=0.423) are the least pronounced. Three pairs of canonical roots were found. The neuroendocrine root of the first pair determines the parameters of immunity by 95.7%, the second pair by 86.5%, and the third pair by 69.2%. The elements of the Thymocytogram (in descending order: epitheliocytes, lymphocytes, endotheliocytes, lymphoblasts, and macrophages), the phagocytic activity of blood neutrophils and monocytes, the relative content of NK- and B-lymphocytes in the blood, as well as the percentage of lymphocytes and macrophages in the Splenocytogram turned out to be most influenced by neuroendocrine factors. Conclusion. There is a close canonical correlation between registered neuroendocrine factors and immunity parameters in general. At the same time, both the severity of the immunomodulatory activity of individual neuroendocrine factors and the subjection to the regulatory influence of individual parameters of immunity differ significantly.

Эффекты пептида GHК и его структурных аналогов D-Ala-GHK и GHК-D-Ala на состояние врожденного иммунитета и перекисного окисления липидов в условиях кожной раны

Objective: to study the effects of the GHK peptide and its structure analogues on the mechanisms of innate immunity and lipid peroxidation in wound. Materials and methods. The experiments were performed on 70 male Wistar rats. The skin wound was modeled by applying a full-layer wound with an area of 250 mm2 on the animal's back. The peptides Gly-His-Lys (GHK), D-Ala-Gly-His-Lys (D-Ala-GHK), and Gly-His-Lys-D-Ala (GHK-D-Ala) were used in doses of 0.5 μg/kg and 1.5 μg/kg, which it was administered intradermally in doses of 0.5 μg/kg and 1.5 μg/kg in a volume of 0.1 ml for 3, 7 or 10 days. The activity of lipid peroxidation (LPO) processes in blood serum was assessed by the content of malonic dialdehyde (MDA) and acylhydroperoxides (AHP). The antioxidant mechanisms were evaluated by determining the activity of catalase and the total antioxidant activity (OAA) of blood serum. Phagocytic activity of blood neutrophils was assessed by phagocytic index (PI) and phagocytic number (PN). The activity of oxygen-dependent mechanisms in phagocytes was evaluated in a spontaneous nitroblue tetrazolium test (NBT). Results. After the administration of GHK, the tendency to PI, PN and completeness of phagocytosis prevailed, which mainly persisted with the use of peptides D-Ala-GHK and GHK-D-Ala. At the same time, the GHK-D-Ala peptide at a dose of 1.5 μg/kg had the most stable effect on phagocytic activity. The data obtained in the NBT are largely consistent with the PN indicators. At the same time, the effects of structural analogues, unlike GHK, were manifested throughout the experiment. Significantly significant changes in the activity of LPO and antioxidant mechanisms were observed with the use of all peptides. However, their dynamics, orientation and severity throughout the experiment were quite complex. Conclusion. GHK and its structural analogues, D-Ala-GHK and GHK-D-Ala, had an effect on the indicators of innate immunity and LPO in a skin wound, the severity and direction of which depends on the healing period. At the same time, the most pronounced and stable effects were observed when using GHK-D-Ala. That demonstrates the importance of protecting the tripeptide molecule from the action of carboxypeptidases.

Effect of allogeneic blood transfusion on neutrophil functional activity and lymphocyte cytotoxicity in recipient rabbits

The relevance of this paper is that transfusion of allogeneic blood to recipient animals is always associated with immunological risks. In this regard, the purpose of this study was to assess the state of phagocytic activity of blood neutrophils by indicators of phagocytic index, phagocytic number, and oxygen-dependent bactericidal activity, as well as to establish changes in antibody-dependent cytotoxic activity of lymphocytes in recipient rabbits during allogeneic whole blood transfusion. Modelling of blood transfusions was performed on five clinically healthy rabbits by intravenous administration of whole blood at the rate of 5.5 ml/kg of body weight. Blood samples were taken from animals on Days 3, 7, and 23 after blood transfusion. Neutrophil populations were obtained from blood samples by centrifugation on a double density gradient of 1.077 and 1.093 Ficoll-Verografin. The absorption activity of phagocytes was determined in a microscopic test. To investigate the oxygen-dependent bactericidal activity of neutrophils, a spontaneous test with nitroblue tetrazolium was performed. Antibody-dependent cytotoxic activity of lymphocytes was investigated by colorimetric method. It was found that after the transfusion of whole blood, the phagocytic activity of neutrophils increases with a simultaneous decrease in their absorption capacity. On Days 3 and 7, the results of the spontaneous test with nitroblue tetrazolium decreased. This indicates inactivation of the oxygen-dependent bactericidal activity of neutrophil granulocytes during the first phase of post-transfusion immunological reactions. On Day 23, there was an increase in the values of the indicators of the spontaneous test with nitroblue tetrazolium, which indicates the activation of the bactericidal properties of phagocytes. It was found that on Day 3, the antibody-dependent cytotoxic activity of lymphocytes significantly decreased relative to the initial state, and on Days 7 and 23, it increased. An increase in the antibody-dependent cytotoxic activity of lymphocytes should be associated with the active synthesis of antibodies of the late phase of the immune response. Consequently, transfusion of allogeneic blood causes an immune response in recipient rabbits, without causing immediate and long-term transfusion reactions (changes in heart rate, respiratory rate, body temperature). The obtained results are of practical value both for scientists and practising doctors who use transfusion of whole blood and its components to animals with acute anaemia, impaired functional activity of blood coagulation factors, parasitic, and oncological diseases

Features of immunological reactivity in Sami women

BACKGROUND: The Sami are a small group of indigenous people of the Russian Federation. The Sami have one of the lowest birth rates among all the peoples of the North of Russia. They often have more health problems than residents of other nationalities in the Murmansk region, due to a high incidence of acute respiratory infections. Extreme climatic and geographical conditions of the Arctic have a negative impact on immunological reactivity.
 AIM: To identify the features of immunological reactivity in Sami women.
 MATERIALS AND METHODS: Forty-nine practically healthy Sami women aged from 21 to 44 years living in the Lovozero Village in the Murmansk region were examined. A control group consisting of 50 practically healthy Russian women of the Arkhangelsk region of the same age range was used. The hemogram, phagocytic activity of neutrophils of peripheral venous blood, and the content of lymphocytes with CD3, CD4, CD8, and CD10 phenotypes were studied by indirect immunoperoxidase reaction (Sorbent, Moscow) and flow cytometry (Beckman Coulter Immunotech, France). Additionally, concentrations of IgM, IgG, IgA, sCD54, and sCD62L (Bender MedSystems, Austria) were determined by enzyme immunoassay, circulating immune complexes (CIC) were determined by the precipitation method. The total amount of microflora, cytogram, phagocytic activity of neutrophils, the sorption capacity of epithelial cells, the content of sIgA and CIC IgG were studied in the separated mucous membranes of the pharynx, intestines, and urine. For comparison between groups, the independent samples t-test or the nonparametric MannWhitney U-test was used depending on the distribution.
 RESULTS: In Sami women, the frequency of deficiency of phagocytic activity of blood neutrophils and mucosal secretions was higher than in the control group. Increased reactions of cell-mediated and antibody-dependent cytotoxicity of lymphocytes were recorded. The frequency of elevated concentrations of CIC was higher in the Sami women than in the control group. A remarkable feature of the Sami immune reactivity was high concentrations of IgA and IgM against a background of low IgG content.
 CONCLUSION: In Sami women, the percentage of actively phagocytic neutrophil granulocytes of peripheral venous blood and mucous membranes was lower than in the control. A distinctive feature of the immunoglobulin profile of the blood serum of Sami women was the low IgG content against the background of increased concentrations of CIC, including immunoglobulins of this class. Sami women had higher levels of leukocyte migration into mucosa-associated mucosal tissue, which was correlated with a decrease in the sorption capacity of epitheliocytes, phagocytic activity of neutrophils, and concentrations of secretory immunoglobulins.

Search for additional tests for immunobiological evaluation of the candidate vaccines against African swine fever.

The spread of African swine fever (ASF) in Eurasia has forced a return to the development of live vaccines based on naturally or experimentally attenuated strains of the virus including those resulting from genetic manipulations. This process includes evaluation of the immunomodulating properties of the vaccines. In this report we provide our assessment of two tests for immunobiological evaluation of a candidate live vaccine against ASF from the attenuated ASF virus (ASFV) strain KK-202: (i) investigation of the effect of the attenuated ASFV strain KK-202 on the protectiveness of the vaccine ASFV strain FK-32/135 and a vaccine against classical swine fever (CSF) from the strain LK-VNIIVViM; (ii) determination of the phagocytic activity of blood neutrophils in pigs inoculated with ASFV strains differing in virulence. A simultaneous or sequential inoculation of attenuated strain KK-202 (seroimmunotype II) and vaccine strain FK-32/135 (seroimmunotype IV) into pigs resulted in the loss of protection against the virulent strain France-32 (seroimmunotype IV). Following the simultaneous or sequential inoculations of the ASFV strain KK-202 and the CSF virus (CSFV) vaccine produced from the strain LK-VNIIVViM, the neutralizing antibody titers against the CSFV observed in the experimental groups (after vaccination and after the challenge infection with the virulent CSFV strain Shimen) were not different from those found in animals of the control group. The phagocytic activity of blood neutrophils was shown to increase from 30% in the norm to 50%–94% depending on the virulence of the ASFV strains inoculated into pigs. The results of this work demonstrate the ability of the attenuated ASFV strains to modulate the development of the cellular link of protective immunity without negative impact on the humoral immune response. The informative value of the described immunobiological tests in vivo and in vitro seems to be a more preferable alternative in comparison to the commonly used in vitro tests, which do not always correlate with the development of protection against ASF.

Intestinal histostructure and immune protection activity of age -1+ carp after consumption of a prebiotic feed additive

Currently, it is important in aquaculture to study and use products and additives, which have positive effects on the digestibility of nutrients by helping to normalize the intestinal microflora, thereby modulating fish immune response. The aim of the study was to assess the effect of a prebiotic based on mannan oligosaccharides produced from the outer walls of Saccharomyces cerevisiae cells on the histological structure, morphometric state of the intestine and activity of cellular and humoral components of nonspecific resistance of age -2+ carp. The experiment was conducted in four analogous ponds, which were stocked with age-1 scaly carp with an average weight of 55–58 g with a stocking density of 1000 fish/ha. Carp had been fed balanced compound feed for 60 days. Experimental groups of fish were fed with a prebiotic as a feed additive in the amount of: 0.025 % – Experiment 1, 0.05 % – Experiment 2 and 0.075 % – Experiment 3. Use of the studied prebiotic did not damage the intestinal histological structure of carp of the experimental groups. The villi height and intestinal crypt depth of carp in Experiment 1 (P < 0.05) and Experiment 2 (P < 0.01) were larger compared to the control group. Experiment 3 showed destructive changes in intestinal mucosa. The results of histological examination indicated the effectiveness of the use of the studied prebiotic in the amount of 0.025 and 0.05 % in the diet of age -1+ carp due to the absence of pathological changes and active morphofunctional state of the intestine, which, in turn, had a positive effect on feed absorption and metabolic processes in fish body. Analysis of the indicators of cellular and humoral links of natural resistance showed that in carp of the second and third experimental groups lysozyme activity of serum was 5 and 11 % (P < 0.001) higher than in the control group. Similar changes, though found in a greater extent, were recorded in the study of bactericidal activity of blood serum, especially in individuals of the second and third experimental groups (by 6.8 (P < 0.01) and 15.2 %). While with carps of the third experimental group, which used a prebiotic drug in the amount of 0.075 %, the content of CEC was 20.5 % higher than in the control group, which indicates an additional antigenic load on the body and is an unfavorable diagnostic factor. At the same time, with the fish of the second experimental group, which received, respectively, 0.05 % of the supplement of the drug “Actigen” to the main diet, probably higher phagocytic activity of blood neutrophils was recorded. A direct dependence of the growth of the phagocytic index and the phagocytic number of neutrophils on the dose of the studied prebiotic was detected. Immunological studies indicated a dose-dependent effect of the prebiotic on the activity of cellular and humoral components of non-specific resistance of carp. Given this, for the effective functioning of innate protective mechanisms, for the increase of the body's immune potential and productivity, it is most appropriate to apply to the diet of carp prebiotic supplement “Actigen” in the amount of 0.025 and 0.05 %.

ЕСТЕСТВЕННАЯ РЕЗИСТЕНТНОСТЬ И РАЗВИТИЕ ЦЫПЛЯТ РЕМОНТНОГО МОЛОДНЯКА ЯИЧНОГО НАПРАВЛЕНИЯ

Blood and lymph make up the circulatory link of the bird's immune system. It is necessary to note the humoral factor of immune protection-lysozyme. The purpose of scientific research is to study the level of natural resistance and development of egg-type chickens after aerosol application of the disinfectant Argodez. Research and production experiments were conducted in the conditions of the poultry farm "snow", Bryansk region, p. Voucher. Aerosol disinfection was performed in the poultry house before settling in with day-old chickens. Test shop were treated once Arcadetm of 0.01%, the rate of 2 ml/m3, and control Desalina - f - 5 ml/m3., with the generator cold fog IGEBA -UNIPRO-5. At the age of 24 hours, two groups of egg - type chickens, the Lohmann-brown cross, were formed using the method of analogous groups. Blood was taken from the underwing vein and by decapitation, before feeding, at the daily, 30-day, 60-day age. Bactericidal, lysozyme, and phagocytic activity of blood serum was determined according to generally accepted methods. Control weighing of chickens of both groups was performed on days 30 and 60 of the study. A significant increase in phagocytic activity of blood neutrophils was found in the experimental group of chickens on the 30th-21.40 % and 60th day of research-25.40 %, as well as a tendency to increase the bactericidal activity of blood serum by 1.90 % and 7.90 % and the lysozyme activity of blood serum by 8.46 % and 9.17 %. It was also found to increase safety by age periods. The highest safety was observed at the age of 1-30 days and was 99.50 %, then 30-60 – 99.10 %.

Влияние пептида GHK-D-Ala на механизмы врожденного иммунитета и процессы перекисного окисления липидов в условиях инфицированной раны

The objective is to study the effects of Gly-His-Lys-D-Ala (GHK-D-Ala) peptide on mechanisms of innate immunity and lipid peroxidation processes in infected wound. Materials and methods. The experiments were carried out on 80 Wistar male rats. The infected wound was modeled by applying a full-layer wound on the back and performing primary wound surgery with primary suturing 24 hours later. GHK-D-Ala peptide was injected at a dose of 0.5 µg/kg into the wound area every day for 30 days. The phagocytic activity of blood neutrophils was studied in spontaneous and stimulated nitroblue tetrazolium reduction test (NBT-test). The activity of lipid peroxidation (LPO) processes in rats’ blood serum was assessed by the content of malondialdehyde (MDA) and acylhydroperoxides (AHP). Results. GHK-D-Ala administration had a positive effect on both phagocytosis mechanisms and the level of LPO products in comparison with the control group. There was a significant increase in NST-positive neutrophils in the spontaneous test against peptide administration on days 3, 7, 10, and 30 (p˂0.001) and in the stimulated NST test on days 3, 7, and 10 (p < 0.001). The functional neutrophil reserve had been significantly increased by day 10 (p˂0.05) and had been significantly lower by day 30 (p < 0.05). Phagocytic index and phagocytic number were significantly higher on days 3 and 10 (p < 0.01). The phagocyte activity index significantly increased on days 3 and 10 (p < 0.05-0.001).A significant decrease in the MDA concentration was demonstrated on days 3, 7, 10, and 30 (p<0,001). There was a significant decrease in the content of AGP on days 3, 7, 10 and 30 (p < 0.001). Conclusion. The corrective effect of Gly-His-Lys-D-Ala peptide on the phagocytic activity of granulocytes and the processes of lipid peroxidation in infected skin wounds was established.

ВСТРЕЧАЕМОСТЬ PNEUMOCYSTIS JIROVECII У ЧАСТО БОЛЕЮЩИХ ДЕТЕЙ ПРИ ПОСТУПЛЕНИИ В РЕСПИРАТОРНОЕ ОТДЕЛЕНИЕ И ПРИ РЕАБИЛИТАЦИИ В САНАТОРИИ

The aim of the study was to determine the frequency of occurrence and to assess the significance of markers of pneumocystosis in frequently ill children (FIC) during inpatient treatment and during rehabilitation in a sanatorium. Materials and methods of research: 115 children with recurrent respiratory diseases were observed: 52 were treated in a hospital and 63 underwent rehabilitation in a sanatorium. Biological samples (oropharyngeal swabs, saliva, feces and blood serum) were examined for pneumocystosis. Laboratory techniques used: a polymerase chain reaction (PCR) to detect DNA pathogens, an indirect immunofluorescence reaction (NRIF) – antigens, and immunoforment analysis (IFA) – specific antibodies of immunoglobulins (Ig) of classes M (IgM) and G (IgG). The material for laboratory research was smears from the posterior pharyngeal wall (PCR, NRIF) and blood serum (IFA). Microbiological studies included seeding, isolation of cultures with quantitative counting of colony-forming units in samples of oropharyngeal swabs, feces. Determination of the functional activity of the oropharynx microbiocenosis by the concentration of short-chain fatty acids in saliva by means of GLC method. Phagocytic activity of blood neutrophils was also investigated. Results: in hospital patients, markers for pneumocystis were found in 60% of cases, including the active form (40%) and pneumocystis (13%); in the sanatorium – 33% of cases, with active infection (26%) and detection of pneumocystis (17%). Inpatients were diagnosed with clinical variants of pneumocystis infection: pneumonia, obstructive bronchitis, acute respiratory infections; schoolchildren have been diagnosed with active pneumocystis infection and carriage. Dysbiotic disorders of the loci of the oropharynx and intestines were identified in all examined subjects, inhibition of the phagocytic activity of blood neutrophils and shifts in the concentration of short-chain fatty acids have been observed. Conclusion: a high proportion of active pneumocystis infection (40% and 27% of cases) was revealed in FIC treated in the hospital and in the sanatorium, which represents a risk group for infection/reinfection with pneumocystosis in the premises.

Supplementation of antioxidant micronutrients reduces stress and improves immune function/response in periparturient dairy cows and their calves

BackgroundPeriparturient period induces stress in cows which fluctuates hormonal and metabolic function and causes immune suppression. Apart from impairing the health, production, and reproduction of cows, it also influences the well-being of newborn calves by decreasing the colostrum quality. Micronutrients are known for optimal health and production and their effects on parturition stress, immune response in both cow and its calf need to be explored. AimThe aim of this study was to see the effect of oral supplementation of micronutrients during the prepartum period on the health status of crossbred dairy cows and subsequently on their newborn calves. MethodsA total of 42 healthy multiparous cows were selected and randomly divided into five groups with seven cows in each group, i.e. control (Basal Diet, BD), VA group (BD + vitamin A, 105 IU), Zn group (BD + zinc sulphate, 60 ppm), VE group (BD + vitamin E, 2500 IU), and combined supplementation (CS) group (BD + combination of VA, Zn, and VE). The supplements were offered in compounded concentrate DM (100 g) to individual cows once daily before the morning feeding and the remaining portion was incorporated in the TMR. Feeding was started one month before the expected days of calving till calving. Blood samples were collected from cows at days -15, -7, -3, 0, +3, +7, and +15 relative to the day of calving. Blood samples from newborn calves and milk samples of cows were collected at days 0, +3, +7, and +15. Milk somatic cell counts (SCC) were estimated using a cell counter. Cortisol was estimated by ELISA kit in blood and milk plasma of cows and in the blood plasma of their calves. Total immunoglobulins (Ig) were estimated in milk of cows and serum of calves using zinc sulphate turbidity method. Blood neutrophils from cows and calves were studied for phagocytic activity (PA) using nitro blue tetrazolium (NBT) assay.Data were analysed by repeated-measures two-way ANOVA using the mixed procedure of SAS, and the pairwise comparison was performed using a multiple comparison test (Tukey). ResultsCombined supplementation of micronutrients decreased (P < 0.05) maternal blood plasma (control vs. CS group, 5.98 ± 0.20 vs. 3.86 ± 0.23 ng/mL) and milk plasma (3.96 ± 0.13 vs. 2.71 ± 0.10 ng/mL) cortisol, milk SCC (3.05 ± 0.11 vs. 2.12 ± 0.10 × 105 cells/mL) and increased (P < 0.05) total milk Ig concentration (18.80 ± 0.11 vs. 23.04 ± 0.57 mg/mL) and the PA of blood neutrophils (0.84 ± 0.03 vs. 1.07 ± 0.03). Similarly, lower blood cortisol concentration (9.69 ± 0.35 vs. 6.02 ± 0.18 ng/mL) and higher (P < 0.05) total Ig (23.26 ± 0.11 vs. 30.34 ± 0.70 mg/mL) and PA of blood neutrophils (0.37 ± 0.02 vs. 0.52 ± 0.02) were observed in the calves born to CS group of cows as compared to the control. Highest (P < 0.05) positive effects (lower stress levels and higher immune response) of treatment were noticed in CS group followed by VE group and then Zn group. However, VA group didn’t differ from the control group. ConclusionOur results indicate that micronutrient interventions during the prepartum period can improve the health status of dairy calves and subsequently the well-being of their calves.

Pharmacological assessment of immunotropic activity of new gel metabiotic on cellular and humoral immunity in experimentally modeled thermal skin burns

Thermal skin burns were experimentally modeled in laboratory animals revealed immunotropic effects of new gel metabiotic consisting of complex biological active substances produced by Bacillus subtilis B-3679 strain as part of three gel transcutaneous bases: tizol, eftiderm and organosilicon glycerohydrogel on cellular and humoral immunity. The aim of the study was to evaluate immunotropic activity of a new gel metabiotic on cellular and humoral immunity in experimentally modeled thermal skin burns in 60 white laboratory Wistar rats, weighed 180-220 g, for metabiotic consisting of a complex biologically active substances produced by Bacillus subtilis B-3679 strain containng the three gel transcutaneous bases: tizol, ephtiderm and organosilicon glycerohydrogel (KTGG). Registered, standardized antimicrobial and wound-healing ointment levosin was used as a control. The experimental data obtained confirm the effectiveness of using the drugs studied for treatment of first and second degree thermal burns revealed by faster decreased area of affected area compared to experimental animals treated with levosin. While evaluating therapeutic effectiveness of experimental samples of metabiotic-based gel preparations, their influence on various parameters of cellular and humoral immunity in modeled thermal skin burn was examined. We assessed phagocytic activity of blood neutrophils and peritoneal macrophages, quantified T and B lymphocytes, as well as antibody-forming cells and immunoglobulins of various classes. It was found that in experimental animals reproducing models of traumatic skin injuries, there is an increase in the level of all the studied parameters of humoral immunity. Concentration of immunoglobulin M, circulating immune complexes and immunoglobulin E, as well as various cytokines, increased larger. The level of α-interferon and the receptor antagonist interleukin-1 was elevated. Thus, the study of the humoral status of experimental animals that received test metabiotic-based gel preparations in modeled thermal skin burns, allows to conclude that they significantly influenced various arms both cellular and humoral immunity. More marked effects were observed for metabiotic-based drugs containing tizol and KTGG. In general, based on the experimental data we can conclude that the relatively high therapeutic effectiveness of examined drugs in the treatment of thermal skin burn wounds in experimental animals, use of which provided earlier removal of necrotic masses, accelerated regeneration processes, and prevented development of secondary wound infection in burns.

In vitro assay for phagocytic activity by canine neutrophils for &lt;i&gt;Escherichia coli&lt;/i&gt;

This study investigates the in vitro phagocytic activity of blood neutrophils of dogs in Sri Lanka for Escherichia coli. The study sample comprised of 70% mongrel and 30% cross bred dogs of both sexes. The leukocyte isolation method described yielded up to 12,930/il viable leukocytes of which 61-90% was neutrophils, with viability ranging from 90-99% (mean 96%). The mean percent phagocytosis (MPP) of canine blood neutrophils was significantly higher at 30 minutes incubation (89%) than at 15 minutes incubation (70 %) with E.coli. The findings support the fact that innate immunity can be measured qualitatively and quantitatively by the technique described which will be useful for the identification of immunocompromised dogs.

ASSESSMENT OF INDICES OF NONSPECIFIC RESISTANCE OF THE ORGANISM IN CONDITIONS OF NARROWING OF THE LUMEN OF THE TRACHEA IN AN EXPERIMENT

The narrowing of the lumen of the trachea due to cicatricial stenoses of the trachea is one of the urgent problems of the modern surgery. The processes taking place in the trachea and lungs lead to a change in the state of the immune system. Assessment of the dynamics of indices of nonspecific resistance is one of the methods for assessing the severity of the course of the disease. We studied the indices of nonspecific resistance of the organism in narrowing the lumen of the trachea in an experiment in the early postoperative period. The study was carried out on the basis of the scientific department of experimental surgery with the vivarium of Irkutsk Scientific Centre of Surgery and Traumatology, on 24 white male Wistar rats with a body weight of 300–350 g. We simulated narrowing the lumen of the trachea by placing a silicone tube in the lumen of the trachea for 14 days. The parameters of phagocytosis, the state of phagocytic activity of blood neutrophils, spontaneous NBT-test, induced NBT- test were studied. The animals were withdrawn from the experiment on the 3rd, 7th and 14th day. We revealed that as a result of narrowing of the lumen of the trachea with the original procedure, the nonspecific resistance of the organism was violated with inhibition of the phagocytic activity of leukocytes. There was inhibition of phagocytic activity of leukocytes and depletion of the functional reserve of leukocytes.

THE ROLE OF IMMUNOLOGICAL DISORDERS IN THE DEVELOPMENT OF ATOPIC DERMATITIS

The basis for the development of atopic dermatitis (AtD) is a complex interaction between genetic disorders, environmental factors, lack of the skin barrier and immunological disorders. In view of the importance of immunological disorders in the etiopathogenesis of atopic dermatitis, the study of these parameters in patients with AtD is a necessary aspect to assess the severity of the course, prognosis of complications and recurrence. Purpose. Comprehensive assessment of clinical and immunological parameters in adolescents with atopic dermatitis. Materials and methods. The study included examination of 28 patients with atopic dermatitis aged from 12 to 16 years, with disease duration from 10 to 15 years, which included analysis of complaints and anamnesis data, evaluation of disease severity on the SCORAD scale, General clinical and immunological studies. The peculiarities of cellular and humoral immunity in the relapse stage were determined. The main indicators of phagocytic activity of blood neutrophils were also investigated. Results. In the analysis of anamnestic data of patients, a history of diseases associated with atopia in 16 (57.1%) of 28 adolescents (AtD, bronchial asthma in relatives of the first and second line of kinship) was revealed. When assessing the disease severity on a scale of SCORAD, prevailed patients with moderate to severe atopic dermatitis over — 19 (67,9%) patients, the average score of the assessment of the severity of AtD amounted to 29.3 ± 1,25. The conducted immunological examination with assessment of cellular and humoral immunity showed significant changes in immunological reactivity in patients with AtD. On the part of the cellular immunity level it is necessary to note a clear decrease in CD3+ and CD8 + , as well as an increase in the subpopulation of T-lymphocytes — CD4 + and b-lymphocytes — CD20 + in most patients compared to normal indicators characteristic of this age group. Compared to healthy adolescents, there were high rates of IRA and NK cells (CD16 + ). In all patients there was an increase in immunoglobulin M and G, a high content of immunoglobulin E. The formation of secondary immunodeficiency, manifested by a decrease in phagocytic activity of leukocytes and incomplete phagocytosis, which is a factor predisposing to the development of pyoderma. Conclusion. Changes in immune status in patients with AtD are expressed by imbalance of population Of T-lymphocytes, increase of various classes of immunoglobulins and considerable oppression of practically all indicators of functional activity of neutrophils of blood.

Inflammatory factors and immunophenotypes in adjustment disorders

To identify inflammatory and autoimmune markers (enzymatic activity of leukocyte elastase (LE), functional α1-proteinase inhibitor (α1-PI), the level of autoantibodies to neurospecific antigens S100b and myelin basic protein (MBP)) as well as phagocytic activity of blood neutrophils of patients with disorders of adaptation, to determine certain immunophenotypes and analyze their possible relationships with disease characteristics. The study included 40 patients with adaptation disorders, mostly women. Diagnostic evaluation and clinical qualification of patients was carried out in accordance with ICD-10: 'Adjustment disorder' (F43.2). The control group consisted of 23 individuals matched for age and sex with patients. The activity of LE and α1-PI was determined by spectrophotometry, and the levels of autoantibodies to S100b and MBP by ELISA, phagocytic activity by the absorptive capacity of neutrophils of peripheral blood of melamine-formaldehyde latex particles. In the total group of patients with adaptation disorders, increased enzymatic activity of LE and functional α1-PI was shown compared to controls (p<0.001 and p<0.0001, respectively). There were no differences in the level of autoantibodies to neuroantigens, and changes in phagocytic index (PhN) compared with the control, however the tendency to reduction of phagocytic number (PhN) was observed. Patients were stratified by leading psychopathological symptoms (predominance of asthenic-depressive or anxious-depressive symptoms, polymorphic symptomatology) and by immunophenotype: (A) inflammatory markers - in the range of control values, (B) - the increase compared to the control activity of both LE and α1-PI, (C) preferential increase in the activity of α1-PI only. The frequency of these immunophenotypes was similar within each of the clinical subgroups. The results suggest the involvement of inflammation in the pathogenesis of adjustment disorders due to stress factors. Various immunological variants differed by proportion of inflammatory markers were not associated with clinical symptoms.

The phagocytic activity of blood neutrophils in patients with chronic generalized periodontitis of I-II severity level on the background of parasitosis

The article presents the results of a study of phagocytic immunity chain in patients with CGP of I-II severity level on the background of enterobiasis, toxocariasis, giardiasis. The aim of this work was to study the nature of changes in the phagocytic immunity chain in patients with CGP of I-II severity level with parasitoses and patients with CGP without parasitoses. Studies were conducted on 540 patients with CGP, I-II severity level with parasitosis - (principle groups). The comparison group consisted of 90 CGP patients, I-II severity level without parasitosis, the control group – of 30 people without periodontal diseases and chronic diseases of other organs and systems. Phagocytic activity of neutrophils was assessed by their ability to absorb the S.aureus bacteria (strain 209), bactericidal capacity (BC) of phagocytes by the S. Nielsen method. Oxygen-dependent activity of phagocytes was studied in the spontaneous and induced zymosan HCT-test. Statistical processing of materials was made using mathematical statistical methods for the data analysis. There was established a reduction in patients with CGP of I-II severity level with parasitosis and in patients with CGP without parasitosis of the absorption and biocidal ability of peripheral blood neutrophils compared with healthy individuals. At I degree of CGP severity in patients without parasitosis, the number of cells, involved in phagocytosis (FI) was decreased on 13.7%, the number of microbes, absorbed by one cell (FN) on 17.3%, biocide – on 76%. As follows from the obtained data FI and FN in this group of patients did not differ from normal values significantly. In patients with CGP of I-II severity level without parasitosis all three indicators studied had a statistically significant difference. FI was reduced in comparison with the norm on 20.5%, FN – on 24.2%, BC – on 152%. In patients with CGP of I-II severity level with parasitosis the significant differences from the control group were observed both at the I degree of the disease, and at II severity level of the disease. At I degree of severity of the disease, reduction of FI in patients with parasitosis was 22.2 28.0%, FN – 32.8 – 39.7%, BC 158 – 170%, at II degree FI – 29.3 – 34.4 %, FF – 41.4 – 53.5%, BC – 238 – 264%. In patients with CGP of I-II severity level with parasitosis the significant differences compared with patients with CGP of I-II severity level without parasitosis were respectively related to the number of absorbed microbes by a single cell (FN) and biocidal capability of neutrophils (BC). Phagocytic index (PI) of patients with CGP of I-II severity level with parasitosis did not differ significantly from those in patients with CGP of I-II severity level respectively without parasitosis. In patients with CGP of I-II severity level with various forms of parasitosis is reduced as the induced red HCT test, as well as spontaneous HCT test. In patients with CGP of I-II severity level without parasitic, the spontaneous HCT test is increased in comparison with both infested patients, and the control group of individuals.

Titanium dioxide stimulates phagocytic activity of blood neutrophils

The aim. Evaluation of the TiO2 effect on the phagocytic activity of peripheral blood neutrophils. Materials and methods. 17 people were examined - 10 people with atopic bronchial asthma (m/f - 5/5, mean age - 38 [28, 48]), verified according to the international criteria, the second group (control) - 7 people - healthy controls (m/f - 3/4, mean age - 23 [21; 26] years). The phagocytic index and neutrophil activity in the NBT-test were calculated for all the examined patients. 0.008% solution of TiO2 (based on 1 mg / kg of body weight) was added to the test-samples, control samples were added with the physiological solution, standard samples - with a suspension of killed Staphylococcus aureus. All the samples were incubated for 30 minutes at 37°C, the ready smears were microscopied with counting the amount of formazan-positive granulocytes in the NST-test and cells that absorbed 1 or more Stf particles per 100 granulocytes. Results. TiO2 stimulated phagocytosis of staphylococcus with blood neutrophils in 53% of the examined patients by 8-30% (p = 0.02). Under the influence of TiO2 activity of phagocytes significantly increased, in comparison with that in spontaneous NST-test (p = 0.0001), but was lower than induction by staphylococcus (p = 0.049). In patients with allergopathology, TiO2 stimulated phagocytosis of staphylococcus and neutrophil activity in the NBT-test was also effective, as was the culture of staphylococcus aureus (31.2% compared to 34.3%). In the healthy group, the TiO2 solution increased the activity of neutrophils in the NST-test, compared with the physiological solution (on average and in 71% of cases by 10-18%), but not so than the culture of staphylococcus (24%, compared to 39, 7%). Among healthy people, there was a correlation between the hereditary of allergy and the neutrophil activity in the NBT-test with TiO2 (0.78). Conclusions. It was found that 0.008% solution of TiO2 stimulated the activity of neutrophils in the NBT-test, both in patients with atopic bronchial asthma and healthy people after 30 minutes of incubation in a thermostat. In patients with allergies, TiO2 increased phagocytosis of staphylococcus, as well as metabolic activity of neutrophils so much as staphylococcus. A strong correlation was found between the hereditary allergies and the increased metabolic activity of blood phagocytes in the NBT-test with TiO2 in healthy individuals.

Фагоцитарная активность нейтрофилов крови у пациентов с хроническим генерализованным пародонтитом I-II степени тяжести на фоне паразитозов

Фагоцитарная активность нейтрофилов крови у пациентов с хроническим генерализованным пародонтитом I-II степени тяжести на фоне паразитозов

EFFECTIVENESS OF APPLICATION OF COMBINED THERAPY OF EARLY ERYTHEMA STAGE OF LYME DISEASE TAKING INTO CONSIDERATION IMMUNOGENESIS FEATURES

Study innate and adaptive immunity in patients with migrating erythema, clinical effectiveness ofcombined therapy using Immunovac vaccine and dynamics of immunologic parame- ters as a result of the therapy. 37 adult patients with migrating erythema were examined. The patients were divided into 2 groups: 1st gr. (14 individuals) - Immunovac by intranasal-subcutaneous method against the background of basic therapy; 2nd gr. (23 individuals) - 200 mg/day doxycycline therapy for 21 days. Phagocytic activity of blood neutrophils; TLRs expression on mononuclear leukocytes of peripheral blood (PBML) and skin cells in foci by flow cytometry with mAT against TLR2, 3, 4, 5, 6, 7, 8, 9 using flow cytometer FC-500; subpopulation composition of peripheral blood lymphocytes; levels of pro-, anti-inflammation and regulatory cytokines in blood sera by EIA method; IgG, 1gM, IgA in blood sera were studied in patients before treatment and 1 month after therapy. A high level of TLR2, 4, 7, 8 on skin cells in foci, TLR2, 4 - on blood cells; low content of CD95+ and CD25+, high-level of serum IL-lb, IL-2 and IL-4, an increase of general IgE level was detected in patients'with migrating erythema. Immunovac facilitated an increase of CD95+ and CD25+, IFN-y synthesis, reduced the level of general IgE in a more pronounced way than basic therapy. Inclusion of Immunovac into therapy of patients With migrating erythema facilitates increase of clinical effectiveness and correlates with correction of immunologic disorders.