Phagocytic Activity Of Peritoneal Macrophages Research Articles

Study of the immunotoxic properties of pegylated hyaluronidase

The development of safe drugs occupies a special place in the pharmaceutical industry. One of the main tasks of its preclinical phase is to evaluate possible toxic effects of the developed drug on the body and on various systems, including the immune system. The aim of our work was to study immunotoxic properties of pegylated hyaluronidase (PEG- HYAL). Material and methods. Mice F1(CBA/C57Bl/6) were divided into subgroups which were intragastric and intraperitoneally administered with PEG-HYAL in different dosages (50, 500, 1250, 2500 and 5000 U/kg). The number of antibody-producing cells in the spleen, the mass and cellularity of central and peripheral immune organs, phagocytic activity of peritoneal macrophages and neutrophils, delayed hypersensitivity reaction (DHR), level of hemagglutinin to sheep erythrocytes, spontaneous and mitogen-induced splenocyte proliferation were determined. Results. PEG-HYAL did not induce DHR, did not suppress phagocyte activity of peritoneal macrophages, at a dose of 50 ED/kg did not significantly affect the hemagglutinin content to erythrocytes, but at a dose of 500 ED/kg did statistically significantly reduce the titer of specific antibodies. When experimental animals were exposed to PEG-HYAL at doses of 50 and 500 U/kg, spontaneous and mitogen-induced proliferation of splenocytes decreased. Conclusions. The PEG-HYAL trial produced results that can be used to substantiate the administration of the PEG-HYAL-based medication.

ОЦЕНКА ИММУНОМОДУЛИРУЮЩЕЙ АКТИВНОСТИ СУХОГО ЭКСТРАКТА ТРАВЫ ТОПИНАМБУРА

. The purpose of the study was to evaluate the immunomodulatory properties of the dry extract of Jerusalem artichoke herb in terms of weight (thymus, spleen), number of cells (thymus, spleen, bone marrow) of the immune system organs, indicators of cellular and humoral immunity in mice on a model of experimental immunodeficiency induced by the cytostatic methotrexate. Materials and methods. The study of the immunomodulatory activity of the dry extract of Jerusalem artichoke herb was evaluated by the ability to restore the mass (thymus, spleen), the number of cells (thymus, spleen, bone marrow) of the immune system organs, indicators of cellular, humoral immunity and phagocytic activity of peritoneal macrophages of outbred mice under conditions of immunosuppression induced by methotrexate, to the level of intact mice, in vivo. Results. Analysis of the effect of the dry extract of Jerusalem artichoke herb in in vivo systems found that the dry extract of the herb Helianthus tuberosum L. had no effect on the mass (thymus, spleen) and the number of cells (thymus, spleen, bone marrow) of the immune system organs, the severity of the delayed-type hypersensitivity reaction, antibody production, phagocytic activity of peritoneal macrophages of intact mice and restored these parameters of the immune system in animals treated with the cytostatic methotrexate to the level of intact animals, which was comparable to the effect of the reference drug "Immunal". Conclusion. The dry extract of Jerusalem artichoke herb can be considered as a promising agent that restores the parameters of cellular, humoral immunity, the phagocytic activity of peritoneal macrophages and the cellular composition of the immune system under conditions of immunosuppression induced by methotrexate, to the level of intact animals, the effectiveness of which is comparable to the effect of the comparator drug "Immunal

Bacteriocin-Producing Lactiplantibacillus plantarum YRL45 Enhances Intestinal Immunity and Regulates Gut Microbiota in Mice.

Bacteriocins production is one of important beneficial characteristics of probiotics, which has antibacterial property against intestinal pathogens and is helpful for regulating intestinal flora. To investigate the impact of bacteriocin-producing probiotics on gut microecology, bacteriocin-producing Lactiplantibacillus plantarum YRL45 was orally administered to mice. The results revealed that it promoted the release of cytokines and improved the phagocytic activity of peritoneal macrophages to activate the immune regulation system. L. plantarum YRL45 was conducive to maintaining the morphology of colon tissue without inflammation and increasing the ratio of villus height to crypt depth in the ileum. The gene expression levels of Muc2, ZO-1 and JAM-1 were significantly up-regulated in the ileum and colon, and the gene expression of Cramp presented an upward trend with L. plantarum YRL45 intervention. Moreover, L. plantarum YRL45 remarkably enhanced the levels of immunoglobulins sIgA, IgA and IgG in the intestine of mice. The 16S rRNA gene analysis suggested that L. plantarum YRL45 administration up-regulated the relative abundance of the beneficial bacteria Muribaculaceae and Akkermansia, down-regulated the abundance of the pathogenic bacteria Lachnoclostridium, and promoted the production of acetic acid, propionic acid and total short-chain fatty acids (SCFAs) in mice feces. Our findings indicated that L. plantarum YRL45 had the potential to be developed as a novel probiotic to regulate the intestinal barrier by altering gut microbiota to enhance intestinal immunity and ameliorate intestinal flora balance.

Effect of Momordicacharantia polysaccharide on immunomodulatory activity in mice.

Momordica charantia polysaccharides (MCPs) have been reported to exert beneficial roles, such as disease healing, in medicine and pharmacy. However, little is known about their effects on immunomodulation. The present study aimed to explore the possible effects of Momordica charantia polysaccharide (MCP) on the immunomodulatory activity of mice lymphocytes. To this aim, male BALB/c mice aged 6-8 weeks were assigned to the following six experimental groups: i) Normal (NG); ii) model (MG); iii) positive (PG); iv) MCP low-dose (MLG); v) MCP medium-dose (MMG); and vi) MCP high-dose (MHG). An immunosuppressive model was established by the intraperitoneal injection of cyclophosphamide in all groups apart from NG. The NG and MG mice were fed with distilled water, whereas the PG mice were administered with levamisole and the MLG, MMG and MHG mice were fed on low, medium and high (100, 200 and 300 mg/kg, respectively) doses of MCP for 21 consecutive days. Subsequently, the mice underwent surgical procedure and were analysed using a range of procedures, including measurement of the thymus index (TI) and spleen index (SI), assessment of the lymphocyte proliferation rate and cell phagocytosis of peritoneal macrophages, lymphocyte proliferation, secretion and mRNA expression of cytokines IFN-γ, IL-6 and IL-12. The mice divided into six groups as mentioned above and treated for 7 days, in the first 6 days, except NG group, mice in each group were desiccated in the abdominal cavity and sensitized by 1% dinitrofluorobenzene (DNFB). On day 6, mice were sensitized with 20 µl DNFB/acetone/olive oil solution behind the right ear and in front of the right ear. Compared with those in the NG mice (not injected with 80 mg/kg cyclophosphamide), the TIs and SIs of the PG, MLG, MMG and MHG mice were increased. In addition, the inhibitory rate of ear swelling and the phagocytic activity of peritoneal macrophages in the PG, MLG, MMG and MHG mice were increased compared with those of MG. Furthermore, the lymphocyte proliferation rate, the secretion and relative mRNA expression levels of cytokines IFN-γ, IL-6 and IL-12 were significantly increased in the PG, MMG and MHG mice compared with those in the NG mice. The results from the present study suggest that treatment with MCP led to an upregulation of the organ indices of immunosuppressed mice, reduced their delayed allergic reaction indicated by the differential cytokine levels, improved the phagocytic activity of peritoneal macrophages, enhanced the proliferation rate of lymphocytes, increased the secretion and expression of IFN-γ, IL-6 and IL-12. Therefore, MCP may improve the immune function of the immunosuppressed mice.

Evaluation of the immunotoxicity and allergenicity of a new intranasal influenza vector vaccine against tuberculosis carrying TB10.4 and HspX antigens.

A new vaccine candidate TB/FLU-05E has been developed at the Smorodintsev Research Institute of Influenza (Russia). The vaccine is based on the attenuated influenza strain A/PR8/NS124-TB10.4-2A-HspX that expresses mycobacterial antigens TB10.4 and HspX. This article describes the results of preclinical immunotoxicity and allergenicity studies of the new vector vaccine TB/FLU-05E against tuberculosis. The experiments were conducted on male CBA mice, С57/black/6 mice, and guinea pigs. The vaccine candidate was administered intranasally (7.7 lg TCID50/animal and 8.0 lg TCID50/animal) twice at a 21-day interval. The immunotoxic properties of the vaccine were assessed in mice according to the following parameters: spleen and thymus weight and their organ-to-body weight ratio, splenic and thymic cellularity, hemagglutination titer assay, delayed-type hypersensitivity test, and phagocytic activity of peritoneal macrophages. Histological examination of the thymus and spleen and white blood cell counts were also performed. Allergenicity of the vaccine was assessed in guinea pigs using conjunctival and general anaphylaxis reaction tests. The results showed that double immunization with the TB/FLU-05E vaccine did not affect the phagocytic activity of peritoneal macrophages, cellular and humoral immunity after immunization with a heterologous antigen (sheep red blood cells), or the organ-to-body weight ratio of immunocompetent organs (thymus and spleen). The vaccine candidate demonstrated no allergenic properties. According to the results of this study, the TB/FLU-05E vaccine is well-tolerated by the immune system and demonstrates no immunotoxicity or allergenicity.

Effect of individual substances isolated from Silene jeniseensis Willd on the state of the main links of immunity at experimental immune deficiency

Introduction. The search, development and introduction of new drugs with an immunotropic effect are one of the priority tasks of modern immunopharmacology. Numerous studies have proven the immunotropic activity of individual substances isolated from medicinal plants (flavonoids, polysaccharides, ecdysteroids, terpenoids, etc.). In the present study, it is of interest to determine the immunomodulatory effect of individual substances isolated from Silene jeniseensis Willd.The aim of the study. Determination of the immunomodulatory activity of individual substances isolated from Silene jeniseensis: flavonoid isoorientin-2”-O-rhamnoside, polysaccharide arabino-3.6-galactan and ecdysteroid 20-hydroxyecdysone under conditions of cyclophosphamide induced experimental immunosuppression.Methods. Experiments were carried out on F1 (CBAxC57Bl/6) mice. Immunodeficiency was modeled by intraperitoneal administration of cyclophosphamide to control group animals in the dose 250 mg/kg once. Experimental groups of mice received the test substances intragastrically once a day for 14 days against the background of immunosuppression in the following doses: isoorientin-2”-O-rhamnoside – 10 mg/ kg, arabino-3.6-galactan – 3 mg/kg, 20-hydroxyecdysone – 3 mg/kg. The effect of substances on cellular immunity was determined in a delayed hypersensitivity reaction, humoral immunity was determined in an antibody formation reaction by local hemolysis according to A.J. Cunningham. The phagocytic activity of peritoneal macrophages was studied in relation to colloidal ink particles.Results. With the introduction of isoorientin-2”-O-rhamnoside, arabino-3.6-galactan and 20-hydroxyecdysone in experimental animals, there was an increase in the index of delayed-type hypersensitivity reaction by 1.3–1.4 times, the absolute and relative number of antibody-forming cells by 1.4–1.7 times, phagocytic index by 1.2–1.5 times compared with the data in the control group, which indicates the leveling of the suppressive effect of cyclophosphamide on cell-mediated immune response, antibody genesis and phagocytosis of macrophages.Conclusion. Isoorientin-2”-O-rhamnoside and arabino-3.6-galactan have the most pronounced immunomodulatory effect. The obtained data allow us to consider the studied substances as promising plant immunomodulators.

The effect of enzymatic hydrolysate of lymphoid tissue of broiler chickens on the morphofunctional state of macrophages

The goal of studying the mechanisms of action of cells that structure the immune system is to obtain additional data about the influence of immunomodulating peptides of natural origin on the possibility of regulating immune processes. The effect of enzymatic hydrolysate of lymphoid tissue of broiler chickens (EHLTBC) on the viability of culture of peritoneal and alveolar macrophages, on the morphological characteristics and functional activity of macrophages has been studied. The effect of EHLTBC on the viability of culture of peritoneal and alveolar macrophages has been investigated. Morphometric characteristics of macrophages under the influence of EHLTBC have been given. The functional activity of peritoneal and alveolar macrophages has been assessed against the background of the introduction of EHLTBC. Enzymatic hydrolysate of lymphoid tissue of broiler chickens at concentrations of 1, 5 and 10 % does not have a toxic effect on the culture of alveolar and peritoneal macrophages. EHLTBC enhances the phagocytic activity of peritoneal macrophages, which enhances their role in the chemotaxis and immune response. The stimulating effect of EHLTBC of poultry on the ability of macrophages to adhere has been revealed.

CYTOKININ FRACTION OF THE Hericium coralloides INCREASES OXIDATIVE METABOLISM OF MURINE PERITONEAL MACROPHAGIES

Aim. This study was aimed to examine influence of сytokinin fraction of basidiomycete Hericium coralloides on the spontaneous and induced phagocytic activity of murine peritoneal macrophages. Materials and methods. Mononuclear phagocyte (macrophage) fraction of peritoneal exudate of C57BL/6 mice was used. Macrophages were incubated under standard conditions at 37 °C, 100% humidity and 5% CO2 for 4 hours. Phorbol 12-myristate 13-acetate (PMA) was added to part of the samples to activate oxidative metabolism. Hericium coralloides was added in two concentrations that were 5 and 10 times lower than the IC50, defined as an antiproliferative effect on colon cancer cells. Incubation with samples was carried out for 2 hours. Results. When adding PMA, Hericium coralloides (0.017 and 0.035 μg/ml) and in the combination of PMA with Hericium coralloides, activation of reactive oxygen species (ROS) in peritoneal macrophages was revealed by 1.37-1.7 times, compared to the spontaneous activity of phagocytes. Conclusions. Thus, the effect of the cytokinin e.tract of the basidium fungus Hericium coralloides was manifested by an increase in the phagocytic activity of peritoneal macrophages as one of the possible mechanisms of immunomodulatory action.

The Role of β-Adrenergic Receptors in the Regulation of the Functions of Innate Immune Cells during Cold Stress In Vivo.

It was found that 10-min cold stress enhanced stimulated production of ROS, while 60-min cold stress increased both spontaneous and stimulated ROS production by peritoneal macrophages. β-Adrenergic receptor blockade leveled the effect of 10-min stress in stimulated cultures and the effect of 60-min stress in spontaneous cultures. None variants of cold stress affected spontaneous and stimulated production of IL-1β. We observed an increase in the production of IL-1β in stimulated cultures from animals subjected to 10- and 60-min stress against the background of propranolol. At the same time, both variants of cold exposure, irrespective of β-adrenergic receptor blockade, stimulated IL-10 synthesis in spontaneous and activated samples. None of the used models of cold exposure affected the phagocytic activity of peritoneal macrophages. Thus, β-adrenergic receptors are directly involved in the regulation of cytokine production and microbicidal potential of macrophages in acute cold stress.

Mesenchymal stromal cells application for experimental genital tuberculosis combination therapy

BACKGROUND: Cell therapy is a promising trend in treating characterized by inflammation and sclerosis diseases such as genital tuberculosis, marked by pelvic inflammatory disease and uterine tube occlusion.
 AIM: Efficacy evaluation of combined mesenchymal stem cells and specific polychemotherapy treatment of the experimental genital tuberculosis.
 MATERIALS AND METHODS: Chinchilla rabbits (n = 27) were divided on 4 groups. Group 1 (n = 6) comprised control group, healthy animals. Group 2 (n = 7) infected with genital tuberculosis, without treatment. Group 3 (n = 7) infected with genital tuberculosis, treated with anti-tuberculous drugs only. Group 4 (n = 7) infected with genital tuberculosis, treated with anti-tuberculous drugs and mesenchymal stem cells. Culture of M. tuberculosis Erdman in dose 107 CFU/0.2 ml was injected under the left uterine tube serosa for the genital tuberculosis modelling. For treatment efficacy evaluation following tests were used: full blood count, blood chemistry, hysterosalpingography, diagnostic laparoscopy, peritoneal macrophage phagocytic activity assessment.
 RESULTS: Infected with genital tuberculosis animals had leukocytosis, elevated C-reactive protein, swelled and occluded uterine tubes. Treated with anti-tuberculous drugs and mesenchymal stem cells rabbits had lower white blood cell count (8.18 1.39 109/L vs 9.32 1.36 109/L, р 0.05) and C-reactive protein (1.1 0.8 mg/L vs 2.2 1.2 mg/L, р 0.01) compared to animal treated with anti-tuberculous drugs only. Animals in group 4 had also increased peritoneal macrophage phagocytic activity. Treated with mesenchymal stem cells animals had unobstructed uterine tubes, stabilized adhesive process within small pelvis with reparative process prevalence.
 CONCLUSIONS: Mesenchymal stem cells combined with anti-tuberculous drugs therapy favors reparative process in uterine tubes in genital tuberculosis.

In Vitro and In Vivo Study of Pandanus conoideus Oil Extract in the Maturation of Mouse Peritoneal Macrophages

Background: Pandanus conoideus (PC) is a traditional medicinal plant that has an interesting effect on inflammatory pathways, especially in the maturation of peritoneal macrophage (PM). PM involve in the pathogenesis of several types of human inflammatory diseases and abdominal cancers which can be targeted as a potential therapeutic strategy. We aim to explore the role of PC oil extract (PCOE) in the maturation of PM which may influence the effectiveness of inflammation.
 Method: This study is a true-experimental in vitro and in vivo laboratory study using CH3 mice. To explore the role of PCOE on PM maturation, experiments were carried out in vitro (detection of nitrite oxide levels, detection of IL-1β levels, analysis of PM phagocytosis and proliferation, as well as flow cytometry analysis of MHC I, MHC II, CD14, and CD68 expression) and in vivo (analysis of PM phagocytosis and flow cytometry analysis of CD14 and CD68 expression). Data were analyzed using analysis of variance (ANOVA) then followed by Bonferroni Post Hoc test to compare the differences.
 Result: PCOE substantially enhanced NO and IL-1β production from mice PM in a dose-dependent manner (p<0.05), upregulates CD68 and CD14 expression on mice and promotes mouse PM phagocytic activity in vitro and in vivo (p<0.01). However, PCOE does not affect the proliferation and MHC I and MHC II expression of mouse PM.
 Conclusion: PCOE has a function in PM maturation by increasing the production of NO and IL-1β, enhancing the expression of CD14 and CD68, as well as promoting PM phagocytic activity.

Immunomodulatory Effects of Extract of Lingzhi or Reishi Medicinal Mushroom Ganoderma lucidum (Agaricomycetes) Basidiocarps Cultivated on Alternative Substrate.

Ganoderma lucidum is a medicinal mushroom exhibiting numerous health benefits primarily based on strong immunostimulatory effects. The study aimed to investigate if there were differences in effects of extracts of commercially (GC) and alternatively (wheat straw) (GA) cultivated G. lucidum basidiocarps on properties of peritoneal macrophages (PM) and monocyte-derived dendritic cells (MoDCs). Differences in immunomodulatory effects of GC/GA extracts were studied. The viability of treated PMs, their adhesive and phagocytic capability, and their capacity to produce reactive oxygen species (ROS) and NO were tested. Immature MoDCs generated from human monocytes were treated with poly I:C (10.0 μg/ml) and loxoribine (34.0 μg/ml), a selective TLR3 and TLR7 agonists, respectively, and with/without GC/GA extract (100.0 μg/ml). The effect of each combination on phenotypic properties, cytokines production by MoDCs, and their proliferation and Th polarizing capacity was studied. GA extract stimulated the metabolic and phagocytic activity of PMs, their adhesion capability, and ability to produce ROS and NO more strongly compared to GC. Both tested extracts significantly increased allostimulatory and Th1 polarization capacity of simultaneous TLR3 and TLR7-activated MoDCs, but GA extract was more effective. The extract of alternatively cultivated G. lucidum basidiocarps increased production of ROS and NO by TLR4 stimulated PMs and upregulated production of certain cytokines as well as allostimulatory and Th1 polarization capacity of MoDCs. GA extract could be a potent immunostimulatory agent for activation of MoDCs with the simultaneous engagement of TLRs, which seems to be a promising strategy for the preparation of DC-based anti-tumor vaccines.

Evaluation of immunotoxicity of the extended-release form of Afobazol

The research of immunotoxicity of extended-release form of Afobazol was conducted on male CBA, C57BL/6 and F1 hybrids (CBA×C57BL/6) mice. Afobazol was administered per os for 14 days in doses of 12 mg/kg and 120 mg/kg. Control group received a placebo. Weight of thymus, spleen and popliteal lymph nodes was not affected by the extended-release form of Afobazol in doses of 12 mg/kg and 120 mg/kg in F1 hybrids (CBA×C57BL/6) mice compared to the control group (p> 0.05). Cellularity of thymus was significantly increased by the extended-release form of Afobazol in dose of 12 mg/kg (p< 0.01 vs control group). Administration of the extended-release form of Afobazol in doses of 12 mg/kg and 120 mg/kg decreased spontaneous chemiluminescence activity of peripheral blood lymphocytes in 2.0 and 2.2 times, in dose of 120 mg/kg level integral chemiluminescence response S was decreased in 2.4 times (p< 0.05 vs control group). Phagocytic activity of peritoneal macrophages and antibody production in F1 hybrids (CBA×C57BL/6) mice were not affected by administration of the extended-release form of Afobazol in doses of 12 mg/kg and 120 mg/kg (p > 0.05 vs control group). 14 days of the extended-release form of Afobazol in doses of 12 mg/kg and 120 mg/kg did not cause any significant change to intensity of delayed-type hypersensitivity reactions (p> 0.05 vs control group). The results of the study allow us to conclude that administration of the extended-release form Afobazol in the range of studied doses does not induce immunotoxicity.

Effect of oral administration of the fresh juice of Bamboo (Bambusa vulgaris) young shoots on enumeration of bone marrow cells, platelets, splenocyte and phagocytic activity of peritoneal macrophages of rats

BackgroundFresh juice of young shoots of Bamboo is a traditionally acclaimed remedy against immune disorders and febrile illnesses. Though, many other therapeutic activities of Bamboo have been investigated, the immunomodulatory activity align with the traditional claim is yet to be scientifically validated. PurposeThe present study investigates the effect of fresh juice of Bambusa vulgaris (FJBV) young shoots, the Sri Lankan variety, on selected immune responses of rats, accompanied by the toxicological and phytochemical profiles. MethodWistar rats (n = 6/group) were orally administered with 3 doses of FJBV - low dose (LD) 0.25 mL/200 g BW (Body Weight), human equivalent dose (HED) 0.5 mL/200 g BW and high dose (HD) 1 mL/200 g BW - once daily for 2 consecutive days according to the traditional treatment practice. Distilled water (DW) and Cyclophosphamide (2 mg/200 g BW) served as the negative and positive controls, respectively. The effect of FJBV on immune cell enumeration, cytokine levels, and phagocytic activity of peritoneal macrophages and serum antioxidant capacities were investigated using standard procedures. Further, the toxicity of FJBV was investigated to corroborate the safety of oral consumption. Chemical profile was also investigated qualitatively. ResultsOral treatment of FJBV for two consecutive days exhibited significant alterations in immune cell counts, phagocytic activity and serum antioxidant levels. Rat BM (Bone Marrow) counts were significantly reduced by both HED and HD (p < 0.05) while, platelet counts were significantly lowered only by HD of FJBV (p < 0.05). The neutrophil to lymphocyte ratio was significantly reduced by the LD of FJBV (p < 0.05), though the Differential WBC (White Blood Cells) counts were not altered by the FJBV oral treatment (p > 0.05). However, splenocyte (SC) counts were significantly increased by the HD (p < 0.05). The FJBV doses tested herein exhibited non-significant alteration in both serum pro and anti-inflammatory cytokine IFN gamma and IL-10, respectively.The phagocytic activity of peritoneal macrophages was significantly lowered by HED and HD (p < 0.05). The acute oral treatment of HD of FJBV for 14 days was found to be safe in terms of hepatic, renal and immune toxicities. Chemical profile revealed the presence of glycosides, reducing sugars, starch, proteins and saponins which may have led to the bioactivities of FJBV. Conclusion: The present study provides affirmation of the traditional claim of the FJBV as an immunomodulatory preparation. Further studies are warranted to explicate the cytotoxic effect of FJBV, which could be a starting material for immunosuppressive drug development.

Role of MP-4 myelopeptide in regulation of antibody production and functional activity of macrophages under stress conditions

Myelopeptides are endogenous low-molecular weight peptides with immunoregulatory activity. Each of these myelopeptides has an individual sequence and plays a role in correction of immune system disorders. In our earlier studies, we have shown immunomodulatory effect of MP-3, MP-5, MP-6 myelopeptides upon functioning of peritoneal macrophages under the conditions of immobilization and cold stresses. The aim of this work was to investigate the effect of MP-4 myelopeptide upon antibody formation, production of reactive oxygen species, and absorptive activity of peritoneal macrophages in mice under the immobilization stress. Materials and methods. The in vivo experiments were carried out in 24 male Swiss mice weighing 17-22 g. Two-or six-hour immobilization stress was produced in the animals. MP-4 was injected intraperitoneally 30 min before the onset of stress, at a dose of 40 μg/kg. The animals were divided into 4 groups: 1, control; 2, stress; 3, stress + MP-4; 4, MP-4 injection. On day +5, cellularity and the number of antibody-forming cells (AOC) in spleen were assessed by the method of local hemolysis in an agarose gel according to Jerne. Production of reactive oxygen species by peritoneal cells was assessed using the luminol-dependent chemiluminescence (LZHL) reaction. The absorption capacity of peritoneal macrophages was assessed using a standard method using a BDFACSCalibur laser flow cytometer. Labeled St. aureus at a final concentration of 108 cells/ml were used as phagocytosis targets. Statistical analysis was performed using the unpaired Student's t-test. Results. It was found that two-hour immobilization stress did not affect the absorption activity of peritoneal leukocytes, whereas six-hour stress suppressed phagocytosis. Injection of MP-4 in stress-exposed animals did not lead to changes in phagocytic activity of peritoneal macrophages. MP-4 did not correct the stress-induced decrease in the number of antibody-forming cells from the spleen during six hours of immobilization, nor stress-induced increase in absolute amounts of AOC under the two-hour immobilization stress. In the group of animals receiving MP-4 and immobilized for two-hours, an increase in the number of nucleated cells was increased, as compared with the control group. Thus, in contrast to the previously studied myelopeptides, MP-4 did not show pronounced immunomodulatory effects upon these parameters.

In Vivo Evaluation of Immune-Enhancing Activity of Red Gamju Fermented by Probiotic Levilactobacillus brevis KU15154 in Mice.

The purpose of this study was to evaluate the immune-enhancing effect of red gamju fermented with Levilactobacillus brevis KU15154, isolated from kimchi, as a biofunctional beverage using mice. Thirty-two mice were used, and after a 2-week feeding, the growth, cytokine and immunoglobulin production, and immune-related cell activation (phagocytes and natural killer [NK] cells) of the mice were evaluated. The red gamju- (SR) and fermented red gamju- (FSR) treated groups had 3.5–4.0-fold greater T-cell proliferation ability than the negative control group. IFN-γ production in the FSR group (15.5 ± 1.2 mg/mL) was significantly higher (p < 0.05) than that in the SR group (12.5 ± 1.8 mg/mL). The FSR group (502.6 ± 25.8 μg/mL) also showed higher IgG production levels than the SR group (412.2 ± 44.8 μg/mL). The activity of NK cells treated with FSR was also greater than that of cells treated with SR but it was not significant (p ≤ 0.05). Further, the phagocytic activity of peritoneal macrophages was higher in both SR and FSR groups than in the control group but was not significantly different (p < 0.05) between the SR and FSR groups. In conclusion, L. brevis KU15154 may be applied in the fermentation of bioactive food products, such as beverages or pharmaceutical industries, to potentially improve immunity.

Immunological and anticancer activities of seleno-ovalbumin (Se-OVA) on H22-bearing mice

Ovalbumin is the main protein component of egg white. Selenium is one of the essential trace elements. In our research, ovalbumin was modified into seleno-ovalbumin. After seleno-modification, the FTIR spectrum of seleno-ovalbumin appeared two new absorption peaks which belonged to the characteristic absorption peaks of Se-O and SeO. Seleno-ovalbumin could reduce the damage of cancer to immune organs, improve the proliferation capacities of T and B lymphocytes, enhance the NK cells cytotoxicity and increase the phagocytic activity of peritoneal macrophages of H22-bearing mice. Besides, Se-OVA could block the cell cycle of solid tumors cells in G0/G1 phase and accelerate the apoptosis of solid tumors cells through mitochondrial pathway.

Chitosan and chitosan nanoparticles as adjuvant in local Rift Valley Fever inactivated vaccine.

The present study aimed to improve the potency of inactivated Rift Valley Fever Virus (RVFV) vaccine using chitosan (CS) or chitosan nanoparticles (CNP) as adjuvants. Chitosan nanoparticles were prepared by ionic gelation method. Rift Valley Fever Virus (RVFV) inactivated antigen was loaded on CS and CNP to form two vaccine formulations, RVFV-chitosan nanoparticles based vaccine (RVFV-CNP) and RVFV chitosan based vaccine (RVFV-CS). Five groups of mice were used in this study, each group was injected with one of the following: phosphate buffer saline (group1 G1), RVFV-CNP (G2), (RVF-CS) (G3), RVFV-Alum based vaccine (RVFV-Alum) (G4) and adjuvant free RVFV inactivated antigen (RVFV-Ag) (G5). The immunization was performed twice with 2 weeks interval. The results showed that, RVFV-CNP vaccine enhanced strongly the phagocytic activity of peritoneal macrophage (PM), neutralization antibodies titer against RVFV and IgG values against RVFV nucleoprotein than other vaccine formulations did. In addition, the RVFV-CNP and RVF-CS vaccines upregulate the gene expression of IL-2, IFN-γ (which promote cell mediated immunity) and IL-4 (which promote humeral immunity), while RVFV-Alum vaccine upregulate the gene expression of IL-4 only. These findings indicated that CS and CNP were comparable to the alum as adjuvant in efficacy but superior to it in inducing cell-mediated immune response and might be a candidate adjuvant for inactivated RVFV vaccine.

Enhancement of Macrophage Function by the Antimicrobial Peptide Sublancin Protects Mice from Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is the major pathogen responsible for community and hospital bacterial infections. Sublancin, a glucosylated antimicrobial peptide isolated from Bacillus subtilis 168, possesses antibacterial infective effects. In this study, we investigated the role and anti-infection mechanism of sublancin in a mouse model of MRSA-induced sublethal infection. Sublancin could modulate innate immunity by inducing the production of IL-1β, IL-6, TNF-α, and nitric oxide, enhancing phagocytosis and MRSA-killing activity in both RAW264.7 cells and mouse peritoneal macrophages. The enhanced macrophage function by the peptide in vitro correlated with stronger protective activity in vivo in the MRSA-invasive sublethal infection model. Macrophage activation by sublancin was found to be partly dependent on TLR4 and the NF-κB and MAPK signaling pathways. Moreover, oral administration of sublancin increased the frequencies of CD4+ and CD8+ T cells in mesenteric lymph nodes. The protective activity of sublancin was associated with in vivo augmenting phagocytic activity of peritoneal macrophages and partly improving T cell-mediated immunity. Macrophages thus represent a potentially pivotal and novel target for future development of innate defense regulator therapeutics against S. aureus infection.

Haloperidol Reduces the Activity of Complement and Induces the Anti-Inflammatory Transformation of Peritoneal Macrophages in Rats.

It is known that psychotropic substances affect the immune system. Unfortunately, chronic antipsychotic administration causes side toxicological effects, associated with oxidative stress. The mechanisms of these effects are still unclear. We investigated the impact of sub-chronic administration of haloperidol (Hal) on parameters of innate immunity and related systems in healthy rats and compared them with Hal content. Hal administration (0.5mg/kg, 3weeks) resulted in two-fold decrease of the activity of the complement system and hemostasis. Hal content correlated with the activity of the complement (r = -0.71), phagocytic activity of peritoneal macrophages (r = 0.78), leukocyte elastase (r = -0.71) and glutathione-S-transferase activity (r = -0.67). Hal fully blocked in vitro PMA-induced iNOS expression in macrophages and changed their morphology to "anti-inflammatory" phenotype. The comparison of in vivo and in vitro data showed that Hal has a direct effect on phagocytic component of innate immunity and an indirect effect on leukocyte elastase and antioxidant enzymes. The results obtained in the present study indicated that Hal significantly affects homeostasis and causes a number of complex biological transformations. Graphical Abstract.