Activity Of Peripheral Blood Leukocytes Research Articles

Изучение состава фенольных соединений, антирадикальной активности и влияния на фагоцитоз сухого водного экстракта шишек сосны обыкновенной

Pine cone is a new promising type of plant raw material for using in a medical practice. Objective: determination of the phenolic compounds composition of pine cones dry aqueous extract, research of its toxicity, antiradical activity and effect on phagocytosis. Materials and methods. The research object was a collection of pine cones gathered in October 2020. A dry aqueous extract was obtained from the cones by extraction with hot water. The phenolic compounds composition of the pine cones dry aqueous extract was determined by ultra-efficient liquid chromatography/MS on a Waters Acquisition chromatograph with a diode-matrix UV detector and a tandem quadrupole MC detector TQD (Waters). To determine the antiradical activity, a reaction with a stable free radical, 2,2-diphenyl-1-picrylhydrazyl (DPPH), was used. The acute toxicity of the pine cones dry extract was researched by the V.V. Prozorovsky express method on white non-linear mice. The phagocytic activity of peripheral blood leukocytes was determined in vivo by a modified method. Results. The phenolic compounds composition of the extract was analysed. The research identified 12 substances present in the extract, representing at least 1% of the total area on the chromatogram. The following procyanidins were found in the pine cones extract: procyanidin B2, procyanidin B3, procyanidin C1, procyanidin D1. Additionally, flavonoids and phenolic carboxylic acids were identified. The pine cones dry extract demonstrated pronounced antiradical activity, comparable to that of ascorbic acid. The acute toxicity determination revealed that the LD50 of the researched compounds is in doses greater than 1260 mg/kg. Upon examination of the effect of pine cone extract on the phagocytic activity of peripheral blood leukocytes, an increase in the phagocytic index was observed, yet no change was noted in the activity of total leukocyte phagocytosis. Conclusion. The dry aqueous extract of pine cones contains biologically active components of phenolic nature, which suggests that further research into its pharmacological activity may be promising.

CsIL-20, a tongue sole interleukin-20, negatively mediates leucocyte activity and antibacterial defense

Interleukin-20 (IL-20), as an essential member of IL-10 family, plays vital roles in mammalian immunological response such as antimicrobial, inflammation, hematopoiesis, and immune diseases. In teleost, the study about immune antimicrobial function of IL-20 is largely scarce. In this article, we revealed the expression profiles and the immunological functions of the IL-20 (CsIL-20) in tongue sole Cynoglossus semilaevis. CsIL-20 is composed of 183 amino acid residues, with seven cysteine residues and a typical IL-10 domain which comprises six α-helices and two β-sheets, and shares 34.4–71.2 % identities with other teleost IL-20. CsIL-20 was constitutively expressed in a variety of tissues and regulated by bacterial invasion, and the recombinant CsIL-20 (rCsIL-20) could bind to different bacteria. In vitro rCsIL-20 could interact with the membrane of peripheral blood leukocytes (PBLs), leading to the attenuation of reactive oxygen species (ROS) production and acid phosphatase activity in PBLs. In line with In vitro results, In vivo rCsIL-20 could obviously suppressed the host immune against bacterial infection. Furthermore, knockdown of CsIL-20 in vivo could markedly enhance the host antibacterial immunity. Collectively, these observations offer new insights into the negative effect of CsIL-20 on antibacterial immunity.

Updated Analysis of a Prospective, Multicenter, Observational Study on the Prevalence of Type 1 Gaucher Disease in Patients with Multiple Myeloma

Introduction. Type I Gaucher disease (GD1) has been associated with increased cancer risk in the International Collaborative Gaucher Group (ICGG) Registry. Particularly, it is well recognized that the risk for multiple myeloma (MM) in affected individuals may be about 9 times higher than expected in the general US populations (Rosenbloom et al, 2022). Pathogenetic relationship between GD1 and MM is still unclear. It was hypothesized that accumulation of glucosylceramide in the macrophages leads to their activation and chronic immune stimulation favouring polyclonal gammopathies. In this scenario plasma cell clone selection could be promoted and subsequent MGUS/MM could derive from it. Diagnosis of GD1 is often inaccurate or delayed since symptoms and signs of disease are nonspecific or underhand. We designed a prospective study to investigate the prevalence of GD1 in a large MM patient population to provide substantial evidences to consider GD1 in the pathogenesis and management of MM. Patients and Methods. This is an observational, prospective, cross-sectional, multicentre study. Twenty-five Italian hematologic centers participated in this study. Due to the lack of data on the effective prevalence of GD1 in MM, the sample size has been determined considering clinically relevant a prevalence of the condition > 0.5% for defining as “high risk” the selected population. Considering an alpha error of 5% and a statistical power of 95%, approximately 1000 patients should beenrolled.. All MM patients are screened by Dried Blood Spots (DBS) sampling technique, that was centralized at the Istituto per la Ricerca e l'Innovazione Biomedica CNR-Palermo. All patients with a positive DBS test undergo genetic test to confirm the diagnosis of GD1. Primary end-point of the study is the prevalence of DBS test positivity in MM patients, which measures glucocerebrosidase activity in peripheral blood leukocytes (normal range 0.2-2.5 nMol/h/m). If the observed prevalence had been significantly higher than predicted, we would have tried to identify the population at higher potential risk of associated GD1. Results. We enrolled 902 patients with a median age of 68 years (range 37-90), 60% of which were male. No patients were Jews, 2 were Asian and 10 were Black, all others were Caucasian. Newly diagnosed and relapsed-refractory MM were 65% and 35%, respectively, whereas SMM and MM were 15% and 85%, respectively. Monoclonal component was IgG in 60%, IgA in 25%, light chain in 15%. Median Ferritin was 310 ng/ml (range 17-1236) and median Alkaline Phosphatase was 75 U/L (range 29-355). Descriptive data of DBS test, enzymatic activity and genetic tests are reported in the Table. In summary, DBS test was found positive in 12/902 (1.33%). Genetic test confirms GBA gene mutations, single heterozygous in 11 patients and double heterozygous in 1 patient. N370S and L444P loci accounted for 50% of GBA gene mutations found. Double-heterozygous patient started replacement therapy for GD1 with imiglucerase. Conclusions. After the enrolment of more than three quarter of the planned patients, the prevalence of DBS test positivity was higher than 0.5% (1.3%). Genetic test confirmed involvement of GBA genes, all but one in heterozygosity fashion. This study allowed to identify one patient with MM and GD1 requiring therapy for both diseases at the same time. The results of this study support further investigations on the implication of the GD1 in the pathogenesis and management of MM. More data on MM patient subgroups at higher risk of associated GD1 may be provided.

IL8 of Takifugu rubripes is a chemokine that interacts with peripheral blood leukocytes and promotes antibacterial defense.

Interleukin 8 (IL8) is a CXC chemokine that plays a crucial role on promoting inflammatory response and immune regulation. In teleost, IL8 can induce the migration and activation of immune cells. However, the biological functions of IL8 are still unknown in Takifugu rubripes. In this study, we examined the biological characteristics of TrIL8 in T. rubripes. TrIL8 is composed of 98 residues and contained a chemokine CXC domain. We found that the TrIL8 expression was detected in diverse organs and significantly increased by Vibrio harveyi or Edwardsiella tarda challenge. The recombinant TrIL8 (rTrIL8) exhibited significantly the binding capacities to 8 tested bacteria. In addition, rTrIL8 could bind to peripheral blood leukocytes (PBL), and increased the expression of immune gene, resistance to bacterial infection, respiratory burst, acid phosphatase activity, chemotactic activity, and phagocytic activity of PBL. In the presence of rTrIL8, T. rubripes was enhanced the resistance to V. harveyi infection. These results indicated that TrIL8 is a chemokine and involved in the activation of immune cells against bacterial infection in teleost.

Oil-producing enterprise

Arterial hypertension is an urgent health problem worldwide causing an increase in temporary and permanent disability, invalidity and mortality due to cardiovascular diseases. Researchers recognize the multifactorial nature of arterial hypertension, but environmental factors are of particular potential importance. The working conditions at oil production enterprises are characterized by a more pronounced influence of these factors which may predispose for early development of disadaptation disorders, functional changes in immune and humoral regulation, and, finally, for increased risk of cardiovascular diseases in people engaged in oil production. The aim of the present work was to study the features of immunity and humoral risk factors of arterial hypertension in hypertensive employees at an oil-producing enterprise. To this purpose, a comparative analysis of lymphocyte subpopulations (CD3+CD4+, CD3+CD25+), markers of apoptosis (CD3+CD95+, TNFR, p53, Bax, Annexin V-FITC+7AAD), phagocytic activity of leukocytes (absolute phagocytosis index), and the levels of vascular humoral factors (nitric oxide and homocysteine) was performed in employees of an oil production enterprise exposed to adverse production factors. The observational group consisted of employees with established episodes of increased blood pressure. A comparison group consisted of individuals without clinical manifestations of cardiovascular disease. As a result of the clinical and laboratory examination of employees at the oil-producing enterprise with arterial hypertension, some functional changes in immune regulation were revealed. This group was characterized by a significantly (p 0.05) decreased CD3+CD4+, and CD3+CD25+ lymphocyte contents, along with increased levels of regulatory CD127 lymphocytes against the comparison group (p 0.05). The workers at an oil production enterprise with arterial hypertension are characterized by decreased (p 0.05) phagocytic activity of peripheral blood leukocytes using the criteria of absolute phagocytosis. We found some signs of inhibited lymphocyte apoptosis (p 0.05), i.e., a decrease in CD95+, TNFR, and p53 over the background values, as well as increased Bax levels over the comparison group (p 0.05). However, the content of TNFR and p53 significantly (p 0.05) exceeded the reference level, regardless of previous arterial hypertension episodes. Development of the high blood pressure episodes among the employees at oi-producing plant showed a significant association (p 0.05) with elevated levels of homocysteine and nitric oxide concentrations which are known to induce endothelial dysfunction, atherogenesis and, hence, a persistent increase in blood pressure.

Pathogenetic therapy of patients with acne, taking into account the metabolic activity of peripheral blood cells

Objective — to increase the effectiveness of treatment of patients with acne, taking into account the features of the metabolic activity of leukocytes and peripheral blood platelets through the use of pathogenetic therapy.
 Materials and methods. 112 patients with acne (65 women and 47 men) aged 16 to 39 years were monitored. The duration of the disease ranged from 6 months to 18 years. 67 (59.8 %) patients were diagnosed with a mild severity of dermatosis, and 45 (40.2 %) — with moderate one. Determination of phospholipids and glycogen content in neutrophils, monocytes and platelets of peripheral blood was performed in 89 observed patients (48 — with mild and 41 — with moderate severity of dermatosis).
 Results and discussion. In patients with acne there is a probable decrease in the level of phospholipids in peripheral blood neutrophils, which is more pronounced in the moderate severity of the pathological process. The obtained data indicate a violation of the structural integrity and inhibition of the energy potential of granulocytes. The content of phospholipids in peripheral blood monocytes was also likely to decrease, more significantly at moderate severity of acne. However, the vector of changes in glycogen levels in these cells had the opposite direction. The results led us to the adjuvant use of platelet­enriched autoplasma which has a wide range of modulating effects.
 Conclusions. It has been found that in patients with acne, there are various vector changes in the metabolic activity of peripheral blood monocytes which are manifested by a decrease in phospholipids caused by an increase in glycogen content, which characterizes the development of intracellular imbalance. The metabolic activity of the platelet pool of peripheral blood in acne remains intact, but there is a redistribution of the number of hemoelements with different saturation of phospholipids and glycogen. The complex therapy of patients with acne using autoplasma enriched with platelets against the background of standardized drugs can significantly improve both short­term and long­term treatment results due to significant correction of the metabolic activity of peripheral blood leukocytes.

Analysis of phagocytic activity of peripheral blood and exudate leukocytes in patients with facial phlegmon

Objective. To assess the phagocytic activity of peripheral blood and exudate leukocytes in patients with facial phlegmon. Currently, relatively little attention is paid to assessing the functional activity of peripheral blood and exudate leukocytes in facial phlegmon, the incidence of which is characterized by an increase in the number of cases and, especially, among patients with comorbid pathology.
 Materials and methods. To study the phagocytic activity of leukocytes using a method based on the assessment of the absorption of formalinized sheep erythrocytes by neutrophils and monocytes, the samples of peripheral blood and exudate were obtained from 18 patients diagnosed facial phlegmon. Blood leukocytes obtained from 29 healthy donors were used as a comparison group.
 Results. It was shown that for patients with phlegmon of the face, an increase in the phagocytic activity of peripheral blood leukocytes is characteristic. There was observed a redistribution of leukocytes according to the number of absorbed objects towards an increase in actively phagocytizing cells 4438 721 per 1 l (in the comparison group 297 67 per 1 l; p 0.05). However, after migration to the foci of the pathological process, leukocytes lost their phagocytic activity and the number of phagocytic cells decreased to 35.0 8.3 % (for peripheral blood leukocytes of the same patients 64.3 5.4 %; p 0.05). Changes in the phagocytic activity of exudate leukocytes affected both neutrophils and monocytes.
 Conclusions. Thus, in patients with phlegmon of the face, changes in the phagocytic activity of blood leukocytes are probably associated with the indirect influence of cytokines, and in the focus of inflammation, leukocytes come under pressure from microorganisms. In such a situation, it is necessary to select an effective immunotropic therapy.

Innate and Adaptive Immunity in Workers of the Main Occupations Exposed to Fine Particulate Matter in Potassium Chloride Production

Background: Workplace air pollution with fine particulate matter in industrial premises contributes to imbalance of nonspecific and specific immunity factors, increasing the risk of developing premorbid conditions in workers. Objective: To study the features of phagocytic activity and subpopulation T-lymphocytes composition in workers engaged in the potassium chloride production. Material and methods: The study was conducted in 2019–2022 within the Research Program of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing on hygiene problems, Clause 2.2.18 “Development of approaches to early diagnosis of production-related diseases using genomic and postgenomic analysis technologies in workers associated with harmful factors of working conditions”, R&D No. 121081900044-4. The observation group consisted of 54 workers of the main occupations exposed, inter alia, to fine respirable particulate matter in potassium chloride production. The comparison group included 67 individuals having no occupational exposure to industrial hazards. We evaluated the phagocytic activity of peripheral blood leukocytes and determined the level of CD25+ and CD95+ differentiation cluster expression on T-lymphocytes by flow cytometry. Results: We measured high airborne concentrations of fine particles at workplaces of mill, centrifuge and filter operators, granulation and drying workers that were 6.6 and 7 times higher than those of PM2.5 and PM10 in the working environment of the administrative staff, respectively. We also observed that the ability of neutrophils to absorb and digest foreign particles was 20 % lower in the workers of the observation group (p = 0.047), while the proportions of CD25+ and CD95+ lymphocytes in them were 30 % and 60 % lower than those in the comparison group, respectively (p = 0.001–0.046). Conclusion: We established the effect of fine particulate matter as a priority workplace air pollutant on innate and adaptive immunity in workers of the main occupations in the potassium chloride production facility. Parameters of innate (phagocytic number) and adaptive (CD25+ and CD95+ lymphocytes) immunity are recommended for use in early diagnosis of immune dysfunction and the development of occupational diseases in workers with inhalation exposure to fine particles.

Метаболічний потенціал лейкоцитарної та тромбоцитарної субпопуляцій клітин периферичної крові та його корекція у хворих на вугрову хворобу

Objective – to increase the effectiveness of treatment of patients with acne, taking into account the peculiarities of the metabolic activity of leukocytes and peripheral blood platelets through the use of pathogenetic therapy. Materials and methods Under our supervision there were 66 patients with acne (39 women and 27 men) aged 18 to 40 years. The duration of the disease ranged from 6 months. up to 14 years old. 42 (63,6%) patients were diagnosed with a mild degree of severity of the course of dermatosis, and in 24 (36,4%) – moderate. In most patients, the pathological process covered two or three topographic areas, that is, it tended to spread. The content of phospholipids and glycogen in neutrophils, monocytes and platelets of peripheral blood was determined by calculating the average cytochemical coefficient (MCC). The control group consisted of 20 healthy individuals. Results and discussion It is proved that in patients with acne there is a probable decrease in the level of phospholipids in peripheral blood neutrophils, which is more pronounced in the moderate severity of the pathological process. The obtained data indicate, respectively, a violation of the structural integrity and inhibition of the energy potential of granulocytes. The content of phospholipids in peripheral blood monocytes was also likely to decrease, more significantly at moderate severity of acne. However, the vector of changes in glycogen levels in these cells had the opposite direction. The results led us to the adjuvant use of platelet-enriched autoplasma, which has a wide range of modulating effects. Conclusions It has been found that in patients with acne, there are various vector changes in the metabolic activity of peripheral blood monocytes, which are manifested by a decrease in phospholipids, combined with an increase in glycogen content, which characterizes the development of intracellular imbalance. It was determined that the metabolic activity of the platelet pool of peripheral blood in acne remains intact, but there is a redistribution of the number of hemoelements with different saturation of phospholipids and glycogen. It is proved that the use of complex therapy of patients with acne using, against the background of standardized drugs, autoplasma enriched with platelets, can significantly improve both short-term and long-term treatment results by significantly correcting the metabolic activity of peripheral blood leukocytes.

CsIL-11, a teleost interleukin-11, is involved in promoting phagocytosis and antibacterial immune defense

Interleukin (IL)-11 is a multifunctional cytokine belonging to the IL-6 family, which plays essential roles in immune response. However, much less is known about the immunological functions of IL-11 in teleost. In this study, we investigated the immune properties of a teleost IL-11 homologue (CsIL-11) from tongue sole Cynoglossus semilaevis. CsIL-11 possesses four conserved α-helices and conserved CsIL-11 receptor binding residues L86 and R187, and shares 23.3%–80.1% identities with other IL-11 homologues. CsIL-11 expression was constitutive in tissues, with most abundant in blood and least abundant in spleen, and upregulated by bacterial challenge in blood, spleen, and head kidney. Recombinant CsIL-11 (rCsIL-11) in the native form of monomer, could bind to peripheral blood leukocytes (PBLs) membrane and enhance the activation and phagocytosis of PBLs. When administered in vivo, rCsIL-11 could markedly promote the host to defend against microbial infection. Overall, our findings show that CsIL-11 plays a pivotal role in regulating PBLs phagocytosis and antibacterial immunity.

A teleost interleukin-16 is implicated in peripheral blood leukocytes recruitment and anti-bacterial immunity

Interleukin-16 (IL-16), as a lymphocyte chemoattractant cytokine, plays a crucial role in regulating cellular activities and anti-pathogen immunity. In teleost, the information about the antibacterial effect of IL-16 is scarce. In our study, we examined the immune functions of an IL-16 homologue (CsIL-16) from tongue sole Cynoglossus semilaevis. The CsIL-16 precursor (proCsIL-16) is comprised of 1181 amino acid residues, sharing 21.1%–67.3% identities with IL-16 precursor from invertebrate and vertebrate. The C-terminal proCsIL-16 containing two PDZ domains was designated as mature CsIL-16 which was released into the supernatant of peripheral blood leukocytes (PBLs). CsIL-16 was expressed in various tissues and regulated by bacterial invasion. Recombinant CsIL-16 (rCsIL-16), as a homodimer, was able to bind to the membrane of PBLs and played essential roles in regulating chemotaxis and activation of PBLs, which in vitro inhibited intracellular survival of E. tarda. Under in vivo condition, rCsIL-16 could dramatically regulate the induction of inflammatory genes, and suppress the bacterial dissemination in fish tissues. Collectively, our results reveal that CsIL-16 plays positive roles in antibacterial immunity, and provide insights into the immune function of CsIL-16.

Опыт внедрения неонатального скрининга болезни Гоше с использованием искусственного флуорогенного субстрата

Важность неонатального скрининга болезни Гоше во многом обусловлена созданием эффективной патогенетически обоснованной терапии. Постановка диагноза строится на обнаружении снижения активности фермента бета-глюкоцереброзидазы в лейкоцитах периферической крови. Newborn screening of Gaucher diseases is important because of the development of treatment options that improve clinical outcome. The diagnosis of Gaucher disease relies on demonstration of deficient glucocerebrosidase enzyme activity in peripheral blood leukocytes.

Analysis of the Leukocyte Response in Calves Suffered from Mycoplasma bovis Pneumonia.

Mycoplasma bovis is known to be a cause of chronic pneumonia in cattle. To date, the disease pathomechanism has not been fully elucidated. Leukocytes play a key role in host antimicrobial defense mechanisms. Many in vitro studies of the effect of Mycoplasma bovis (M. bovis) on leukocytes have been performed, but it is difficult to apply these results to in vivo conditions. Additionally, only a few studies on a local immune response in M. bovis pneumonia have been undertaken. In this study, the experimental calf-infection model was used to determine the effect of field M. bovis strains on changes of the peripheral blood leukocyte response, including phagocytic activity and oxygen metabolism by cytometry analyses. An additional aim was to evaluate the lung local immunity of the experimentally infected calves using immunohistochemical staining. The general stimulation of phagocytic and killing activity of peripheral blood leukocytes in response to the M. bovis infection points to upregulation of cellular antimicrobial mechanisms. The local immune response in the infected lungs was characterized by the T- and B-cell stimulation, however, most seen in the increased T lymphocyte response. Post-infection, strong expression of the antigen-presenting cells and phagocytes also confirmed the activation of lung local immunity. In this study—despite the stimulation—both the peripheral and local cellular antimicrobial mechanisms seem to appear ineffective in eliminating M. bovis from the host and preventing the specific lung lesions, indicating an ability of the pathogen to avoid the host immune response in the M. bovis pneumonia.

A microencapsulated feed additive containing organic acids, thymol, and vanillin increases in vitro functional activity of peripheral blood leukocytes from broiler chicks

During the first week after hatch, young chicks are vulnerable to pathogens as the immune system is not fully developed. The objectives of this study were to determine if supplementing the starter diet with a microencapsulated feed additive containing citric and sorbic acids, thymol, and vanillin affects in vitro functional activity of peripheral blood leukocytes (PBLs). Day-old chicks (n = 800) were assigned to either a control diet (0 g/metric ton [MT]) or a diet supplemented with 500 g/MT of the microencapsulated additive. At 4 D of age, peripheral blood was collected (100 birds per treatment), and heterophils and monocytes isolated (n = 4). Heterophils were assayed for the ability to undergo degranulation and production of an oxidative burst response while nitric oxide production was measured in monocytes. Select cytokine and chemokine mRNA expression levels were also determined. Statistical analysis was performed using Student t test comparing the supplemented diet to the control (P ≤ 0.05). Heterophils isolated from chicks fed the microencapsulated citric and sorbic acids, thymol, and vanillin had higher (P ≤ 0.05) levels of degranulation and oxidative burst responses than those isolated from chicks on the control diet. Heterophils from the supplemented chicks also had greater (P ≤ 0.05) expression of IL10, IL1β, and CXCL8 mRNA than those from control-fed chicks. Similarly, nitric oxide production was significantly (P ≤ 0.05) higher in monocytes isolated from birds fed the supplement. The cytokine and chemokine profile in monocytes from the supplement-fed chicks showed a significant (P ≤ 0.05) drop in IL10 mRNA expression while IL1β, IL4, and CXCL8 were unchanged. In conclusion, 4 D of supplementation with a microencapsulated blend made up of citric and sorbic acids, thymol, and vanillin enhanced the in vitro PBL functions of degranulation, oxidative burst, and nitric oxide production compared with the control diet. Collectively, the data suggest feeding broiler chicks a diet supplemented with a microencapsulated blend of citric and sorbic acids, thymol, and vanillin may prime key immune cells making them more functionally efficient and acts as an immune-modulator to boost the inefficient and undeveloped immune system of young chicks.

Anti-chemotactic activity in the secretory/excretory products of Lepeophtheirus salmonis

The ectoparasite, Lepeophtheirus salmonis (Kroyer 1837), is effective at avoiding elimination from its host, Atlantic salmon, Salmo salar L., by inhibiting the recruitment of immune cells to the site of attachment. In other ectoparasitic arthropods, numerous factors have been identified that bind or neutralize chemokines preventing their interaction with receptors on the surfaces of immune cells. To determine if L. salmonis is utilizing a similar mechanism of immune modulation, the chemotactic activity of peripheral blood leukocytes (PBL) to leukotriene B4 (LTB4) and the secreted/excreted products (SEPs) of the sea louse were investigated in vitro. The results showed that incubation of LTB4 with SEPs reduced leukocyte migration compared to LTB4 immune stimulation alone. Data suggests that one of the mechanisms L. salmonis may be using to regulate immune cell recruitment in Atlantic salmon is by inhibiting or neutralizing the activity of chemokines.

Tongue sole (Cynoglossus semilaevis) interleukin 10 plays a negative role in the immune response against bacterial infection

Interleukin-10 (IL-10) is a pleiotropic cytokine and plays a crucial role in immunity. In the current study, we examined the expression patterns and biological functions of tongue sole Cynoglossus semilaevis IL-10 (CsIL-10). CsIL-10 is composed of 186 amino acid residues and shares 46.3%–71.7% identities with other teleost IL-10. Csil-10 expression occurred in multiple tissues and was regulated by bacterial infection. Recombinant CsIL-10 (rCsIL-10) in the form of a dimer bound to a wide range of bacterial species but did not affect bacterial growth. rCsIL-10 could interact with peripheral blood leukocytes (PBL) and significantly reduce the phagocytic activity, ROS production, and apoptosis of PBL. When injected in vivo, rCsIL-10 significantly suppressed the expression of proinflammatory cytokines and promoted bacterial dissemination in tongue sole tissues. Consistently, knockdown of Csil-10 significantly inhibited bacterial infection in tongue sole. Taken together, these results indicate that CsIL-10 plays a negative regulatory role in the immune response against bacterial infection.

Influence of Endogenous Interferon Inducer on Phagocytic Activity of Peripheral Blood Leukocytes

Influence of Endogenous Interferon Inducer on Phagocytic Activity of Peripheral Blood Leukocytes

INCREASED LEVELS OF INTERLEUKIN-10 EXPRESSION COMPARED TO INTERLEUKIN-6 IN LEUKOCYTES OF HEALTHY SUBJECTS. COULD IT BE USEFUL IN THE FUTURE OF THE DEPRESSION DIAGNOSIS?

It is a well known, that the physical activity promotes mental health. Physically active people relatively rarely suffer from psychosomatic and depressive disorders. It is possible that the differences in gene activity in peripheral blood leukocytes may be associated with depression, especially genes participating in inflammatory response. Thus, the aim of our study was to investigate the levels of IL6 and IL10 mRNA and IL10/IL6 ratio in peripheral blood leukocytes in healthy, physically active individuals. One hundred healthy young men (20-23 years old) participated in this study. All of them declared regular physical activity. Participants were non-smokers, and consumed alcohol occasionally. To access genes expression, 2 ml of venous blood was collected. RNA isolation was performed and then the relative expression of IL6 and IL10 was calculated using quantitative real-time polymerase chain reaction (Q-RT-PCR). Pro and anti-inflammatory balance was calculated as 2^relative of IL10/ 2^relative expression of IL6. Low expression of tested genes was found in healthy young men. Mean expression for IL6 was 2^0.051 (n=90) and for IL10 2^0.08 (n=98). Mean ratio IL10/IL6 was 2^1.58. Higher expression of IL10 compared to IL6 may be essential not only for the physical performance but also for the mental health. Diverse reports in the literature may be associated with choosing various control groups, i.e. sedentary or older individuals. It is possible, that measuring the expression of IL6 and IL10 (especially the ratio IL10/IL6) in peripheral blood leukocytes may be useful in the assessment of depressive disorders. Thus, molecular study of active young men can confirm the need for physical activity among people suffering from depression, but further studies are needed, particularly among people with psychosomatic disorders.

IMMUNIZATION WITH PROTEINS OF PSEUDOMONAS AERUGINOSA OprF AND aTox ENHANCES THE PHAGOCYTIC AND BACTERICIDAL ACTIVITY OF LEUKOCYTES IN MICE

Aim. To study the effect of vaccine preparation against Pseudomonas aeruginosa on the phagocytic and bactericidal activity of immunocompetent cells in mice. Materials and methods. Preparation: 25 pg of OprF, 50 pg of aTox sorbed by 75 pg of aluminum hydroxide. For immunization, the recombinant protein preparation was mixed in equal weight fractions with an aluminum hydroxide gel, diluted in phosphate buffered saline, and sorbed for 12 hours at 4°C. The vaccine preparation was administered intraperitoneally in 0.5 ml to BALB/с mice. The phagocytic activity of peripheral blood leukocytes was determined from the absorption capacity of heat-killed FITC-labeled Staphylococcus aureus by neutrophils and monocytes of immunized mice by flow cytometry. The bactericidal activity of mice blood leukocytes was assessed for the live culture of S.aureus using flow cytometry Cytomix FC-500 (Beckman Coulter). Results. Administration to mice of recombinant proteins P. aeruginosa OprF and aTox sorbed on aluminum hydroxide led to an increase in the phagocytic and bactericidal activity of monocytes and granulocytes of peripheral blood. The maximum increase in the number of phagocytized monocytes was observed on the 7th, and granulocytes on the 17th day after the first immunization. Booster immunization did not lead to additional stimulation of phagocytic activity, but the number of phagocytic cells was significantly (p

The effect of intermittent hypoxia under different temperature on the immunomodulation in Streptococcus agalactiae vaccinated Nile tilapia (Oreochromis niloticus)

Dissolved oxygen (DO) and temperature are the potential immunomodulators in fish and play the important roles in regulating immunity. We studied the effect of intermittent hypoxia under different temperature on the immunomodulation in vaccinated Nile tilapia (Oreochromis niloticus). The expression of immune-related genes, enzymatic activities, histology, cumulative mortality, and S. agalactiae clearance were assessed. Study conditions were intermittently hypoxic (4.0 ± 1.0 mg/L DO) at 30 ± 0.5 °C or 35 ± 0.5 °C. Interleukin-1beta (IL-1β), tumor necrosis factor alpha (TNF-α) and gamma interferon (IFN-γ) mRNA expression in spleen and head kidney were significantly lower in vaccinated hypoxic fish compared to the vaccinated normoxic fish. Levels of heat shock protein 70 (HSP70) in tissues showed an opposite tendency. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities were significantly lower in vaccinated hypoxic fish. Malondialdehyde levels were significantly greater under hypoxic conditions. In vitro studies evaluated the effects of intermittent hypoxia at different temperatures on cells of vaccinated O. niloticus. Phagocytic activity of peripheral blood leucocytes (PBLs) and intracellular reactive oxygen species (ROS) production in head kidney cells were significantly decreased by intermittent hypoxia at either 30 °C or 35 °C, while nitric oxide levels in tissues cells increased significantly under hypoxic conditions. These changes were well reflected by the further suppression modulation on S. agalactiae clearance in vaccinated O. niloticus and higher cumulative mortality by intermittent hypoxia. Taken together, intermittent hypoxia at either 30 °C or 35 °C could suppress immunomodulation in vaccinated Nile tilapia.