Percentage Of Phagocytosis Research Articles

Сравнительная оценка показателей иммунного профиля у детей в условиях воздействия бенз(а)пирена и холодового фактора в исследовании «случай-контроль» с моделированием и верификацией полученных результатов в эксперименте in vivo

The comparative assessment of immune profile indicators in children under = benzo(a)pyrene and cold factor influence in a case-control study with the modeling and verification of obtained results in an in vivo experiment was performed. In the course of a case-control study in children under airborne benzo(a)pyrene exposure the signs of phagocytic leukocyte activity inhibition and specific haptenic hypersensitivity formation (IgG to benzo(a)pyrene) associated with blood contamination with benzo(a)pyrene were established. Revealed changes in the immune profile and xenobial biomedia composition in children under airborne benzo(a)pyrene exposure at a dose of 0.00761 mcg/(kg*day) in the subarctic urbanized area, it is comparable to its level in children living under the conditions of external environmental benzo(a)pyrene exposure at a dose of 0.08689 mcg/(kg*day) in an urbanized area in a temperate climate zone (p>0.05). The results of the experimental study of the immunotropic effects of oral benzo(a)pyrene exposure at an average daily dose of 0.175 mcg/kg*day and cold factor influence (15.1±2.6 °C) in vivo also show the signs of phagocytosis inhibition in mice (OR=7.33 - 16.20, p<0.05). At the same time, the minimum values of phagocytosis percentage, phagocytic number and phagocytic index are observed with the combined benzo(a)pyrene and cold factor effect which confirms the hypothesis of immunotoxic benzo(a)pyrene exposure effect modification by a cold factor Keywords: benzo(a)pyrene; cold stress; children; immune profile; phagocytosis; in vivo experiment

Effect of the injection of protoscolex antigens for hydatid cysts on some immunological parameters and the protection of injury

This study aimed to that capability of preparation of antigens from protoscolex found inside the hydatid cyst and likewise possibility of preparing antigen providing protection of the injury of this cysts. Cystic echinococcosis is the more important parasitic disease caused by a cestode tapeworm of species Echinococcus granulosus, spreading a lot in Iraq and the Eastern Mediterranean Countries in addition to its world distribution. The protoscolex found inside hydatid cysts results from the germinal layer of hydatid cyst isolated from people by the surgery section at the Fallujah teaching hospital to find out the extent of its influence on both of the cellular and humeral immunity. 20 white mice (Balb/c) were divided into two groups, the first group of white mice (Balb/ c) was injected subcutaneously with a concentration of 0.1ml of antigen and it measured antibodies IgG, IgA, IgM, IgE as well as injected with complement C3, C4 and the insulation of neutrophil cells and measuring percentage of phagocytosis, reduction of a dye NBT and viability of neutrophil cells and in the second group, the protoscolex was injected in the liver with a concentration 0.1ml to find out the extent of prevention provided by the antigen to prevent or to reduce injury. The results showed a significant increase for all types of antibodies and complements C3 , C4 and reduction of a dye NBT and phagocytosis especially at the time 60 minute and the results were not showing significant differences on viability of neutrophil cells. As for prevention of injury, the results have shown a significant difference at the level < 0.05 and reached the percentage of prevention 56.4 % compared to the subordinate control. The results showed small size of the hydatid cyst in the experience parameters compared to the subordinate control.

Phagocytosis in Marine Coccolithophore Gephyrocapsa huxleyi: Comparison between Calcified and Non-Calcified Strains.

Coccolithophores play a significant role in marine calcium carbonate production and carbon cycles, attributing to their unique feature of producing calcareous plates, coccoliths. Coccolithophores also possess a haplo-diplontic life cycle, presenting distinct morphology types and calcification states. However, differences in nutrient acquisition strategies and mixotrophic behaviors of the two life phases remain unclear. In this study, we conducted a series of phagocytosis experiments of calcified diploid and non-calcified haploid strains of coccolithophore Gephyrocapsa huxleyi under light and dark conditions. The phagocytosis capability of each strain was examined based on characteristic fluorescent signals from ingested beads using flow cytometry and fluorescence microscopy. The results show a significantly higher phagocytosis percentage on fluorescent beads in the bacterial prey surrogates of the non-calcified haploid Gephyrocapsa huxleyi strain, than the calcified diploid strain with or without light. In addition, the non-calcified diploid cells seemingly to presented a much higher phagocytosis percentage in darkness than under light. The differential phagocytosis capacities between the calcified diploid and non-calcified haploid Gephyrocapsa huxleyi strains indicate potential distinct nutritional strategies at different coccolithophore life and calcifying stages, which may further shed light on the potential strategies that coccolithophore possesses in unfavorable environments such as twilight zones and the expanding coccolithophore niches in the natural marine environment under the climate change scenario.

Effects of Increasing Levels of Purified Beta-1,3/1,6-Glucans on the Fecal Microbiome, Digestibility, and Immunity Variables of Healthy Adult Dogs.

Yeast-purified beta-1,3/1,6-glucans (BG) can modulate dogs' immune systems and microbiome, but the optimal inclusion dose remains unknown. The aim of the study was to evaluate the effects of 0.0, 0.07, 0.14, and 0.28% inclusion of BG in a dry extruded diet on the digestibility, immunity, and fecal microbiota of healthy adult dogs. Eight male and female border collies [n = 4; body condition score (BCS) = 5] and English cocker spaniels (n = 4; BCS = 5), aged 3.5 ± 0.5 years, were randomly distributed into two 4 × 4 balanced Latin squares. Fecal microbiota (using 16S rRNA sequencing, Illumina®), apparent digestibility coefficients (ADC) of nutrients, fecal concentrations of short-chain fatty acids (SCFA) and branched-chain fatty acids (BCFA), ammoniacal nitrogen, lactic acid, IgA and pH, lymphocyte immunophenotyping, intensity and percentage of phagocytosis and oxidative burst were determined. No differences were observed in Faith (p = 0.1414) and Pielou-evenness (p = 0.1151) between treatments, but beta diversity was different between 0.0% and 0.14% BG groups (p = 0.047). Moreover, the Firmicutes phylum was the most abundant in all groups and exhibited the highest relative abundance after the consumption of 0.14% BG, a finding considered beneficial for the canine microbiome. The Erysipelotrichaceae and Ruminococcaceae families, along with the Faecalibacterium and Prevotella genera, considered favorable for their involvement in butyrate production and other metabolites, showed increased abundance after the consumption of 0.14% BG. The potentially pathogenic Proteobacteria phylum displayed lower abundance after the consumption of 0.14% BG. Fecal concentrations of the evaluated compounds and pH did not differ after consumption of the BG at all percentages. Higher crude protein ADC was found after 0.14 and 0.28% BG consumption (p < 0.0001), but no differences were found for other nutrients. Phagocytosis, oxidative burst, and lymphocyte populations were not modulated by any of the treatments; however, 0.14% BG modulated the lymphocyte T CD4+:CD8+ ratio (p = 0.0368), an important marker of immune system efficiency. The inclusion of 0.14% BG resulted in the best responses and was the best dose evaluated.

Immunomodulatory Activity of Robusta Green Coffee Extract on Macrophage Phagocytosis and Lymphocyte Proliferation in Vitro

Robusta green coffee contains a large quantity and variety of polyphenols and flavonoids which are known to have antioxidant and anti-inflammatory properties. This research to investigate the immunomodulatory activity of Robusta green coffee extract on macrophage activity stimulated with lipopolysaccharide (LPS) and lymphocyte proliferation. Macrophages were isolated from mice peritoneal tissue, and Lymphocytes were isolated from lymph organs. The immunomodulatory activity on macrophages was evaluated based on Active Phagocytes Cells (APC) and the proliferation of lymphocytes was evaluated by the MTT Assay which was previously given extracts with varying concentrations (20 µg/ml, 40 µg/ml, 50 µg/ml, 60 µg/ml, 70 µg/ml), then the data were analyzed by ANOVA (α=0.05). The aqueous extracts of Robusta green coffee significantly affects macrophage activity and lymphocyte proliferation in vitro. The extract was decrease the percentage of phagocytosis (PP) macrophage stimulated with LPS, however there was increase phagocytosis capacity (PC) and lymphocyte proliferation stimulation index is 3, illustrating the extract has a low immunomodulatory effect on lymphocyte proliferation. Based on these results, Robusta green coffee extract holds potential as an immunomodulatory agent, affecting against macrophage phagocytosis and lymphocyte proliferation.

Features of the immune status and comorbidities in children with alopecia areata

Increased incidence of alopecia has been noted in children at the present time. Participation of autoimmune (immunopathological) mechanisms in pathogenesis of this disease necessitates additional study of immune status and characteristics of comorbid pathologies. The aim of our study was to specify the features of immune status and comorbidities in children with alopecia areata. The observation group consisted of children with various types of alopecia areata (n = 57), a comparison group included children without clinical manifestations of alopecia or a history of alopecia (n = 157). We performed a comparative evaluation of major lymphocyte subpopulations (CD3+, CD3+CD4+, CD3+CD8+, CD3+CD19+), interleukins (IL-4, IL-6), immunoglobulins (IgA, IgM, IgG), parameters of phagocytic activity (absolute phagocytosis, percentage of phagocytosis, phagocytic number and phagocytic index) and IgE to house dust and cat hair. Analysis of comorbidities was also performed. Statistical processing was carried out with Jamovi software. We have found that the focal clinical form of alopecia prevailed over the subtotal and total forms by 1.8 times (p = 0.033) in the observation group rather than in comparison group, with common variable immunodeficiency being more often (1.4-fold), chronic tonsillitis (3.9-fold), allergic rhinitis (3.9-fold) and autoimmune thyroiditis, which was absent in the comparison group. Among the children with alopecia, disturbances of the T-cell link were revealed, i.e., a higher median levels of the relative and absolute numbers of CD3+CD4+ cells (p = 0.001 to 0.003), larger proportions of elevated values for IgA (4.1-fold), IgM (7.3-fold), IgG (13.2-fold) with p-levels of 0.0001 to 0.0008; increased IL-4 (8.1-fold) and IL-6 (4.6-fold), with p = 0.002-0.004, along with medium and relatively strong correlations with alopecia. In children with alopecia, we have determined a 3.3-fold proportion of reduced values of absolute phagocytosis and 3.7-fold reduced percentage of phagocytosis (p = 0.0012 to 0.028), with an sufficient correlation (weak to moderate strength) with alopecia, as well as lower values of the median phagocytic index and phagocytic number in the observation group (p &lt; 0.001) associated with a average-strength correlation. Hence, an imbalance of the immune system components was revealed in children with alopecia which manifested with signs of immune hyperfunction, characteristic, e.g., of autoimmune and allergic processes, accompanied by more frequent registration of autoimmune thyroiditis and allergic rhinitis. Moreover, distinct signs of immune deficiency, are found, characterized by a decrease in phagocytic activity and higher incidence of common variable immunodeficiency and chronic tonsillitis.

Immune function, egg quality and production responses in layer hens fed two different lignocellulose fibre supplements during the early laying period

The effects on immune function and egg productivity of 2 commercial lignocellulose supplements with similar insoluble and soluble fibre concentrations and water holding capacities, were determined in Hy-Line Brown hens from 20 weeks-of-age for 8 weeks. Hens were fed 1 of 3 diets (6 pens/diet, 3 hens/pen): Control diet – a commercial layer diet; Diet LC1 - Control diet plus 1.5 g Arbocel RC fine/100 g diet; Diet LC2, Control diet plus 1.5 g OptiCell C5/100 g diet. After 4 weeks, heterophil phagocytosis index and percentage (innate immunity), was greater in LC1 (p < 0.05) than Controls. After 8 weeks, absolute and relative weights (g/100 g BW) of LC1 thymus glands were heavier (p < 0.05) than Control; numbers of intestinal Peyer’s patches in LC1 were greater (p < 0.05) than LC2 and Control; total and relative areas of LC1 Peyer’s patches were higher (p < 0.05) than Control. Yolk immunoglobulin Y concentrations of LC1 were higher but not significantly, than Control and LC2. Between 6 and 8 weeks, egg weights and eggs produced/hen per d of LC1 hens were greater (p < 0.05) than Controls. Feeding LC1, but not LC2, during early lay significantly improved immune function and egg productivity compared to Control. As dietary fibre concentrations of LC1 and LC2 were similar, differences in their relative concentrations of chemical components such as cellulose and lignin or types of polysaccharides, may have contributed to the improvements during early lay.

Immunomodulatory properties of plant polyphenols shown in an &lt;i&gt;in vitro&lt;/i&gt; experimental model

Polyphenols exert a wide range of biological effects, including immunomodulatory action. Studying the effects of flavonoids on phagocytic activity of specialized phagocytic cells seems to be a rather promising direction for their further usage as pharmacological (therapeutic) agents. Quercetin and luteolin are the most commonly studied flavonoid substances with pleiotropic action. In-depth study and understanding of immunotropic mechanisms (e.g., regulation of phagocytosis) is a prerequisite for adequate pharmacotherapy in infectious conditions, nonspecific inflammatory diseases, autoimmune and oncological disorders. The aim of our work is to study the effect of flavonoids upon phagocytic activity of professional phagocytes (neutrophils) using an in vitro test system. The biological material (venous blood) from 30 practically healthy people (adults n = 15, children n = 15) was used in the present work. The study was carried out in accordance with established international regulations. 0.5 mg/L of Luteolin (basic substance content ≥ 98%) and Quercetin (basic substance content ≥ 95%) were added to experimental samples and incubated for 20 min at 37 °С. The percentage of phagocytosis, phagocytic number (mean number of formalin-treated sheep erythrocytes engulfed per one neutrophil) was determined in control and experimental samples using light microscopy. The unidirectional nature of phagocytosis inhibition by quercetin and luteolin was noted in the test experiments. A statistically significant (p &lt; 0.05) decrease in the phagocytosis intensity by 10% was shown in experimental blood samples obtained from adult patients compared with control values. When quercetin and luteolin were added to blood samples obtained from children, a statistically significant (p &lt; 0.05) decrease in phagocytosis by 30% was noted against control values. At the same time, the mean percentage of phagocytosis and phagocytic number in blood samples after the addition of flavonoids were found to be in the range of reference values, thus suggesting adequacy and physiological suppression of excessive activities of innate immunity compartments by quercetin and luteolin. At this concentration, the flavonoids were found to exert a more pronounced suppressive effect on phagocytic activity in children. Modeling of immune response using the phagocytosis indices assayed in experimental in vitro models with neutrophils from practically healthy adults and children enables us to expand the knowledge of mechanisms underlying the immunotropic effects of flavonoids (quercetin and luteolin), in order to correct immunopathological conditions.

Effect of radiotherapy on phagocytosis percentage and index in patients with oral squamous cell carcinoma.

Phagocytosis plays an important role in the fundamental process of immunity and maintains systemic tissue homeostasis. Phagocytosis function is assessed in radiotherapy to signify the prognosis of patient. Therefore, we designed a study to assess and compare the phagocytosis percentage and phagocytic index of the leucocytes in patients with oral cancer before and after radiotherapy. A total of 60 subjects of both genders, between 50 and 75 years old, were included in the study. The subjects were categorized into three groups: group I: oral squamous cell carcinoma (OSCC) patients before radiotherapy (n = 20), group II: OSCC patients after radiotherapy (n = 20), and group III: control (n = 20). Further, 2-mL blood samples were collected from the patients during the 1st day before radiotherapy and at the end of radiotherapy. The obtained samples were centrifuged, separated leucocytes were incubated with C. albicans, and smears were made to determine the phagocytosis percentage and phagocytic index. The obtained values were tabulated and statistically analyzed. P < 0.05 was considered to be statistically significant. Phagocytosis percentage and phagocytic index of OSCC patients before radiotherapy, OSCC patients after radiotherapy, and in the control group and the difference was found to be statistically highly significant (P < 0.01). Phagocytosis function can be determined by assessing the phagocytic index and percentage to know the effect of radiotherapy-induced systemic immune suppression and predict the prognosis of OSCC patients after radiotherapy.

Immunomodulatory Activity of Begonia Medicinalis Ethanolic Extract in Experimental Animals

Benalu batu (B. medicinalis) is a plant endemic to Central Sulawesi that has been reported to possess anticancer, antioxidant, and antiviral activities. The mechanism of action of these activities is still unclear. This study aimed to evaluate the immunostimulatory effect of ethanol extract of B. medicinalis on an in vivo model by measuring the macrophage phagocytotic activity and cytokine production of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) on male mice and rats, respectively. The extract was obtained by the maceration method for 3 × 24 h using 70% ethanol. The doses of the extract were 60, 120, and 240 mg/kg body weight (bw). The percentage of macrophage phagocytosis and the TNF-α, and IFN-γ levels were measured on the eighth day, one hour after intraperitoneal injection of Staphylococcus aureus ATCC 25923. The result showed that ethanol extract of B. medicinalis activated macrophage phagocytosis in a dose-dependent manner, with a significant increase in cytokine expression of TNF-α and IFN-γ. The optimal dose was 240 mg/kg bw, with a higher percentage of phagocytic activity and higher levels of TNF-α and IFN-γ than Stimuno® as a positive control and other dosage treatments. This study suggests that B. medicinalis ethanol extract has the effect of an immunomodulator and provides a scientific basis for traditional usage of this herb plant.

The effect of endomorphins-1, -2 on the production of reactive oxygen species and the absorption activity of innate immunity cells &lt;i&gt;in vitro&lt;/i&gt; and &lt;i&gt;in vivo&lt;/i&gt;

Background. A high percentage of people in the world suffer from acute or chronic pain, the treatment of which is based on the introduction of morphine and its derivatives, agonists of the µ-opioid receptors, leading to the development of addiction in patients. An urgent task is to study the properties of endogenous opioid peptides, which are capable of showing not only pronounced analgesic activity, but also have a number of other effects on the periphery, while having a much smaller spectrum of side effects. Of particular interest are the immunomodulatory effects of peptide agonists of µ-receptors – endomorphins, tetrapeptides, the effects of which are not described in detail in the literature. Due to their structure and properties, endorphins can be considered as potential substitutes for low-molecular-weight opiates. The aim. To investigate the effect of endorphins-1,2 on the synthesis of oxygen radicals and the absorption activity of innate immunity cells in vitro, in vivo. Materials and methods. The object of the in vitro study is peripheral venous blood leukocytes from donors aged 22–40 years; the object of the in vivo study is peritoneal flush cells of Swiss white male mice. Oxygen-dependent microbicidal activity was evaluated using a luminol-dependent chemiluminescence reaction. Phagocytic activity was evaluated by flow cytometry. Results. It has been found that endomorphins inhibited spontaneous production of reactive oxygen species in vitro, in vivo. Endomorphin-2 reduced the intensity of respiratory explosion in stimulated cultures of peritoneal macrophages. Blockade of opiate receptors with naloxone in vitro, in vivo did not cancel the effects of endomorphins. Both peptides and naloxone had no effect on the phagocytic activity of macrophages of the peritoneal cavity of mice in vivo, but in vitro endomorphins led to an increase in the percentage of phagocytosis of peripheral blood leukocytes. Conclusions. The effects of endomorphins in vitro, in vivo are predominantly depressive.

Heightened immune surveillance in Drosophila melanogaster populations selected for faster development and extended longevity

Maximization of life-history traits is under constraints due to both, limitations of resource acquisition and the restricted pathways of resource allocation. Drosophila melanogaster has served as an excellent model organism to not only unravel various trade-offs among life history traits but also numerous aspects of host immune response. Drosophila larvae are semi-aquatic that live, feed and excrete inside the food source-often over-ripe fruits and vegetables that are rich in both commensal and pathogenic microbiota that can impact the larval survival. In this study, we have used six populations of D. melanogaster, three of which are selected for faster pre-adult development and extended adult longevity, and their three ancestral controls, to explore the impact of selection on the basal immune activity in the larval stage. The larvae from selected populations had nearly significantly upregulated plasmatocyte density, significantly higher percent phagocytosis, phagocytic index and higher transcript levels of Tep3, eater and NimC1. Selected populations also had significantly upregulated crystal cell number along with higher transcript of PPO2. Out of seven tested AMPs level, Drosomycin was significantly upregulated in selected populations while Drosocin was significantly higher in control populations. ROS levels were comparable in the selected and control populations. Our results strongly suggest that enhanced basal immune activity during larval stage manages the faster development and could be responsible for comparable larval survival of selected and control populations.

Plasma steroids and immune measures vary with restraint duration in a toad (Rhinella icterica)

Immunoenhancing effects have been widely described following acute stressors in several vertebrates, and valuable contributions have been made from studies on acute stress to understand hormonal-immune interactions. However, most studies focus on hormonal and immune responses after standardized time lapses, neglecting potential influence of duration of exposition to stressor. Herein, we investigate fluctuations of plasma hormone concentrations (corticosterone and testosterone) and immunity (neutrophil to lymphocyte ratio, phagocytosis of blood cells, and plasma bacterial killing ability) in a toad species (Rhinella icterica) in response to six different periods of exposure to restraint stress. We observed increased plasma corticosterone concentrations following restraint in all sampled times (0.5 to 48 h), with the highest values being observed during the first hour (0.5 to 1 h). Restraint-induced increases in the neutrophil to lymphocyte ratio and phagocytosis percentage were observed from the first 0.5 h, gradually increasing after that with the time of restraint. We also observed decreased testosterone plasma concentrations in response to a more prolonged restraint (24 and 48 h). No changes were observed in plasma bacterial killing ability following restraint. Together, our results demonstrate dynamic time-related hormonal and immune changes. These results point to the fact that for some species measuring hormonal and immune variables at single time points following a stressor might work better when preceded by a study of the temporal changes of the response variables to the stimuli applied. Also, time of response needs to be considered when different variables are used as proxies of stress.

Immunostimulant Activity of Pegagan Embun Herbs Extract (Hydrocotyle sibthorpioides Lam.) With Carbon Clearance Method Towards Male White Mice

Pegagan embun (Hydrocotyle sibthorpioides L.) has been used traditionally to cure various diseases such as anti-inflammatory, neutralizing toxins, dysentery, and others. This study aims to determine the immunostimulant effect of pegagan embun herbs extract using the carbon clearance method, the number, and the percentage of leukocyte cells. In this study, 25 male white mice were used, which were divided into five groups, groups 1, 2, and 3 given pegagan embun herbs extract in a dose of 10 mg/kgbw; 50 mg/ kgbw; 200 mg/kgbw, group 4 was given 0,5% Na CMC, and group 5 were given stimuno 50 mg/kgbw. The preparation is given orally for seven consecutive days. On the eighth day, the phagocytosis index, number, and percentage of leukocyte cells were determined. The results showed that pegagan embun herbs extract has immunostimulant activity. The phagocytosis index of the 10 mg/kgbw dose group is 1.0211, 1.27796 for the 50 mg/kgbw, and 1.5250 for the 200 mg/kgbw. Then, the number of leukocytes of the 10, 500, and 200 mg/kgbw are 8.820/μL, 9.906/μL, and 10.790/μL, respectively. The percentage of lymphocyte cells are 58,5%, 60,6% and 62,8%, and for segment neutrophil cells were 26%, 27,6% and 30% for group 10, 50, and 200 mg/kgbw extract. Based on the results, that the pegagan embun herbs extract has immunostimulant activity against male white mice.

Features of the immune profile and polymorphism of candidate genes in children population of an industrially developed region with excessive contamination of the biological medium with mercury

Introduction. Analyzing the negative impact of technogenic chemicals on the health of the children’s population of industrially developed regions is an urgent problem of preventive medicine. Excessive accumulation of mercury in the human body causes disadaptation changes in the immune regulation of physiological processes. Therefore, the analysis of the features of the immune profile associated with polymorphic variants of candidate genes as markers of early disorders of the child population’s health status is relevant in preserving the health of the population of industrialized regions. Materials and methods. A clinical and laboratory examination of the biological environment of 215 children aged 4-6 years was carried out. The observation group consisted of 133 people living in an industrially developed region. The comparison group consisted of 82 people living in a relatively clean territory. The level of contamination of the biological medium with mercury was determined by inductively coupled plasma mass spectrometry. Identification of CD3+CD4+ -, CD3+CD8+- and CD19+ - lymphocytes was performed by flow cytofluorometry. The study of the phagocytic activity of leukocytes was carried out using formalized ram erythrocytes. The level of IgG production was determined using radial immunodiffusion by Mancini, specific IgG to mercury was carried out using allergosorbent testing with an enzyme label. Identification of single-nucleotide polymorphic variants (SNP) of the GSTA4 (rs3756980), FOXP3 (rs3761547), MTR (rs1805087), TERT (rs10054203) genes was carried out by real-time PCR. Results. Children living near the territory of the chemical industry enterprise in conditions of mercury exposure at a level not exceeding hygienic standards are characterized by an increased level of mercury contamination of urine, exceeding the reference level and the level of the comparison group by 1.8 times (p&lt;0.05). The immune profile of children in the observation group is characterized by a decrease in the CD4+/CD8+ immunoregulatory index due to the decline of CD3+CD4+ helpers and hyperproduction of CD3+CD8+ cytotoxic lymphocytes, inhibition of the phagocytic activity of leukocytes (percentage of phagocytosis, phagocytic number, phagocytic index) against the background of an increase in CD19+ lymphocytes, serum IgG and a marker of specific sensitization - IgG to mercury (p&lt;0.05). Changes in the immune profile of children with an increased level of mercury contamination are associated with the C-allele and TC-heterozygous and CC-homozygous genotypes of the GSTA4 gene (rs3756980), the C-allele and CC-genotype of the FOXP3 gene (rs3761547), the A-allele and AA-genotype of the MTR gene (rs1805087) (OR&gt;1, p&lt;0.05), the G-allele and GG-genotype of the TERT gene (rs10054203) (p&lt;0.05). These genes are responsible for the features of detoxification processes, immunoregulation and longevity programs. Conclusion. The established features of cellular (decrease in CD4+/CD8+ due to CD3+CD4+ deficiency with simultaneous increase in CD3+CD8+, inhibition of phagocytosis) and humoral (hyperproduction of IgG, specific IgG to mercury, CD19+) immunity associated with polymorphic variants of the glutathione S-transferase GSTA4 (rs3756980), transcription factor FOXP3 (rs3761547), MTR (rs1805087), TERT telomerase (rs10054203) in children with excessive contamination of the biological medium with mercury, a complex of immune and genetic markers of the effect and sensitivity of mercury exposure is formed.

The Relevance of CD20 Targeting of Oncolytic Measles Virus in the Treatment of Adult Acute Lymphoblastic Leukemia

The Edmonston-B, vaccine strain of measles virus (MVNSe) has oncolytic activity in numerous preclinical tumour models with safety and some efficacy being demonstrated in phase 1 clinical trials for multiple myeloma and cutaneous non-Hodgkin lymphoma.Adult acute lymphoblastic leukemia (ALL) is an aggressive malignancy where long term survival is achieved in only 50% of patients. The GRAALL study showed that anti-CD20 monoclonal antibody, rituximab, improved outcome in ALL and the results of the UKALL14 trial are eagerly anticipated in this regard. To determine whether targeting MV to the CD20 antigen can recapitulate some of these benefits we used a MV expressing a single chain Fv antibody against CD20 as a C-terminal extension of the H glycoprotein. We also used targeted MV ‘blinded’ to the native MV receptors previously cloned by site-directed mutagenesis. The mechanisms of MV oncolysis remain incompletely understood and our experiments were particularly designed to help clarify the role of the innate immune system.First, we retrovirally transduced CD20 negative B-precursor ALL cell line NALM6 cells to express human CD20. The cells were flow sorted to give ‘high’ and ‘low’ expressing populations based on mean fluorescence intensity. Viral replication was measured by TCID50 at consecutive 24 hourly time-points and MV-N mRNA/GADPH by RQ-PCR showing similar growth curves and MV-N mRNA levels for the targeted and parent virus. Oncolytic activity was measured by trypan blue exclusion at 24, 48, 72 and 96 hours post infection (hpi). Infection of CD20 negative NALM6 cells by MVHaCD20 virus showed similar levels of killing to the parent virus MVNSe, and there was significantly greater levels of killing by MVHaCD20 of the CD20 positive cells when compared to MVNSe (p =0.03). Since it is also possible for the MVHaCD20 to enter cells via native MV receptors CD46 and CD150 (SLAM) we investigated the effect of ‘blinding’ the virus to these receptors. As expected, there was a small, non-significant reduction in efficacy in the CD46 blinded CD20 virus, MVHaCD20CD46blind, but no difference with the CD150 blinded virus, MVHaCD20SLAMblind, as CD150 is not expressed on NALM6 cells.Our previous work has shown that neutrophils are important effectors in mediating MV oncolysis. To further investigate specific mechanisms, we carried out a FACS-based neutrophil phagocytosis assay in which target cells were labelled with PKH67 and the percentage phagocytosis determined by the frequency of dual expressing CD15+/PKH67+ cells relative to the total number of CD15+ cells. These assays demonstrated that in the presence of anti-MV antibody a similar level of phagocytosis was seen 24 hpi with MVNSe, at a multiplicity of infection of 1.0, to uninfected CD20 expressing NALM6 cells in the presence of rituximab (53% c/w 58%). The percentage phagocytosis strongly correlated with the anti-MV IgG concentration from individual donors' serum (Pearson's correlation, r =0.937; p <0.001) The level of phagocytosis was significantly increased when complement was active (student t-test, p =0.008). There was no increase in cell death as indicated by DAPI staining.To further investigate the role of complement we carried out complement cytotoxicity (CDC) assays at 2hpi and 24hpi. As CD46 is a negative regulator to complement the CD46 blind viruses were included in the comparison. The results showed CDC to be a minor mechanism at 24hpi causing 5-10% cell death which was less effective than rituximab which resulted in 13-17% cell death. There was no additional effect with our ‘blinded’ viruses.In conclusion, CD20 targeted MV has increased oncolytic efficacy in-vitro compared to the native MV when infecting CD20 expressing NALM6 cells. Our results suggest that ADCP is a mechanism of MV oncolysis however a recent study in chronic lymphocytic leukemia (Valgardsdottir et al Blood 2017) has indicated that this effect is ‘trogocytosis’ whereby the neutrophils take up small amounts of the cell membrane rather than engulfing the entire leukemic cell. In our study complement was implicated both in increasing the efficacy of this mechanism and also in CDC. Although this requires further exploration in ALL, it remains that neutrophils appear to be an important effector cell of MV oncolysis in the presence of anti-MV antibody. Overall our data suggest that targeting MV to CD20 could have some efficacy in the treatment of ALL. DisclosuresFielding:Baxalta: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees; Amgen: Membership on an entity's Board of Directors or advisory committees.

LPS-induced immunomodulation and hormonal variation over time in toads.

The inflammatory response is a complex process that relies on interactions among multiple endocrine and immune modulators. Studies incorporating time-related and integrative endocrine and immune responses to an immune challenge might shed light on the characterization of the phases of the inflammatory response in anurans. The present study investigated time-related changes (1, 3, 6, and 18 h post-challenge) in plasma corticosterone (CORT), melatonin (MEL) and testosterone (T) levels, phagocytosis percentage (PP), plasma bacterial killing ability (BKA), and neutrophil to lymphocyte ratio (NLR) following a lipopolysaccharide (LPS) immune challenge in Rhinella diptycha toads. Our results showed the response to LPS injection was characterized by increased CORT, PP, BKA, and NLR, with a concomitant decrease in plasma MEL and T. Increased CORT was more pronounced at 6and 18 h, while increased NLR was observed only 18 h post-LPS injection. Meanwhile, plasma MEL and T decreased independently of the time post-LPS injection. Additionally, toads in better body condition showed higher BKA and PP in the LPS-treated group, regardless of the time postinjection. Our results show that toads (R. diptycha) were sensitive to the LPS challenge, mounting an inflammatory response, which started quickly (after 1 h) and developed over time and was influenced by body condition. These results demonstrate a time-related hormonal and immune variation as a consistent pattern of activation of the immune system, as well as of hypothalamic-pituitary-adrenal/interrenal and immune-pineal axes following an immune challenge more deeply studied in mammals, suggesting the evolutionary conservation of the regulatory mechanisms for tetrapod vertebrates.

Encephalitozoon cuniculi takes advantage of efferocytosis to evade the immune response.

Microsporidia are recognized as opportunistic pathogens in individuals with immunodeficiencies, especially related to T cells. Although the activity of CD8+ T lymphocytes is essential to eliminate these pathogens, earlier studies have shown significant participation of macrophages at the beginning of the infection. Macrophages and other innate immunity cells play a critical role in activating the acquired immunity. After programmed cell death, the cell fragments or apoptotic bodies are cleared by phagocytic cells, a phenomenon known as efferocytosis. This process has been recognized as a way of evading immunity by intracellular pathogens. The present study evaluated the impact of efferocytosis of apoptotic cells either infected or not on macrophages and subsequently challenged with Encephalitozoon cuniculi microsporidia. Macrophages were obtained from the bone marrow monocytes from C57BL mice, pre-incubated with apoptotic Jurkat cells (ACs), and were further challenged with E. cuniculi spores. The same procedures were performed using the previously infected Jurkat cells (IACs) and challenged with E. cuniculi spores before macrophage pre-incubation. The average number of spores internalized by macrophages in phagocytosis was counted. Macrophage expression of CD40, CD206, CD80, CD86, and MHCII, as well as the cytokines released in the culture supernatants, was measured by flow cytometry. The ultrastructural study was performed to analyze the multiplication types of pathogens. Macrophages pre-incubated with ACs and challenged with E. cuniculi showed a higher percentage of phagocytosis and an average number of internalized spores. Moreover, the presence of stages of multiplication of the pathogen inside the macrophages, particularly after efferocytosis of infected apoptotic bodies, was observed. In addition, pre-incubation with ACs or IACs and/or challenge with the pathogen decreased the viability of macrophages, reflected as high percentages of apoptosis. The marked expression of CD206 and the release of large amounts of IL-10 and IL-6 indicated the polarization of macrophages to an M2 profile, compatible with efferocytosis and favorable for pathogen development. We concluded that the pathogen favored efferocytosis and polarized the macrophages to an M2 profile, allowing the survival and multiplication of E. cuniculi inside the macrophages and explaining the possibility of macrophages acting as Trojan horses in microsporidiosis.

Ethanol Extract of Spondias pinnata Leaves Reduce Parasite Number and Increase Macrophage Phagocytosis Capacity of Mice Infected by Plasmodium berghei

BACKGROUND: Currently, there is no vaccine against malaria in humans, the development of resistance to anti-malarial drugs, causing the need to find new alternatives to overcome malaria infections. This study aimed to determine effect of Spondias pinnata in increasing cellular immunity, especially phagocytosis activity of peritoneal macrophages against Plasmodium berghei infection.METHODS: This was an experimental study with two stages of research, each stage requires 36 Balb/c mice, aged 2 months and weight 20-25 grams. After one week of acclimatization, the mice were put into 6 different groups, each group consisted of 6 mice. The negative control was a group of mice given distilled water for 14 days then infected by P. berghei in the 15th day. Meanwhile, T1, T2, T3, T4 and T5 groups were given S. pinnata leaves ethanol extract with dose of 25, 50, 100, 200 and 400 mg/kg body weight (BW)/day, respectively, and then infected by P. berghei in the 15th day.RESULTS: The results showed that the lowest parasitemia and the highest capacity of macrophage to phagocytose latex was found in treatment group T3 that received 50 mg/kg BW of S. pinnata leaves ethanol extract. Based on analysis of the Pearson correlation test, there was a significant correlation between percent phagocytosis and parasitemia (p&lt;0.05).CONCLUSION: Ethanol extract of S. pinnata leaves lower the parasite number of P. berghei in Balb/c mice and increase the capacity of macrophage to phagocytose latex. However, the mechanisms of how S. pinnata leaves extract in activating phagocytosis capacity and reducing parasitemia still need further investigation.KEYWORDS: phagocytosis, Plasmodium berghei, parasite number, Spondias pinnata

Water hyacinth (Eichhornia crassipes) leaves enhances disease resistance in Channa punctata from Vibrio harveyi infection

BackgroundChanna punctata, Indian spotted snakehead, has a great economic value in south and south-east Asia being an important protein source for humans. Fish cultures are affected due to various bacterial and viral infections. Vibrio harveyi is a fish pathogenic bacteria which causes several outbreaks throughout the world and leads to huge mortalities. In this study, leaves of Eichhornia crassipes (water hyacinth) were used to investigate its immunostimulatory potential in Channa punctata.ResultsThe immunostimulatory effects of water hyacinth leaves were studied in fish fed with 2.5% and 5% supplementary feed (experimental groups) in comparison to normal feed (control groups). Gas chromatography mass spectrometry (GC-MS) analysis of E. crassipes methanol extract showed presence of various components which have immunostimulatory, antioxidant, antibacterial, and anti-inflammatory activities. The antibacterial activity, antioxidant potential, and presence of phenol and flavonoids in methanol and ethanol extracts supported its use in fish feed. The healthy acclimatized fish were challenged with V. harveyi weekly. Liver function tests, alkaline phosphatase levels, and immunoglobulin content in the experimental groups were improved with respect to those in the positive control group. The spleen and head kidney were obtained at the final day of experiment, and macrophages were isolated; higher percentage of phagocytosis and phagocytic index indicated enhanced cell-mediated immune response in fish due to supplemented feed.ConclusionPlant-infused feed with leaves of E. crassipes can be recommended as a regular feed supplement to enhance fish immunity and disease resistance against the V. harveyi infection.