Activity Of Murine Peritoneal Macrophages Research Articles

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.

Anti-inflammatory effects of the protein hydrolysate and peptide fractions isolated from Salvia hispanica L. seeds

ABSTRACTThe aim of this work was to evaluate the potential anti-inflammatory effect of protein hydrolysate and peptide fractions from Salvia hispanica L. seeds. Protein isolate was obtained using defatted flour of mucilage-free seeds. Hydrolysis process was conducted by enzymatic digestion. The hydrolysate was fractionated using ultrafiltration membranes to obtain the peptide fractions (<1, 1-3, 3-5, 5-10, and >10 kDa). Protein derivatives were evaluated by in vitro activation of murine peritoneal macrophages. The anti-inflammatory activity was determined as NO production, H2O2 release and pro- and anti-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-10) production. All the peptides exerted an anti-inflammatory activity, but peptide fraction between 1 - 3 kDa showed the highest anti-inflammatory effect. This fraction was evaluated on in vivo murine models of TPA-induced ear edema and DNFB-induced delayed-type hypersensitivity, exhibiting inhibitory effects. Hence, the results demonstrated that protein derivatives from S. hispanica L. seeds have in vitro and in vivo anti-inflammatory effects.

Influence of carrageenan on cytokine production and cellular activity of mouse peritoneal macrophages and its effect on experimental endotoxemia.

The in vivo effect of κ/β-carrageenan isolated from the red alga Tichocarpus crinitus on cytokine synthesis and cellular activity of murine peritoneal macrophages and also the protective effect of polysaccharides in LPS-induced endotoxemia in mice was studied. It was established that κ/β-carrageenan given orally at a dose of 100 mg/kg stimulates the induction of anti-inflammatory cytokines (IL-10) in mouse blood cells by more than 2.5-fold compared with control, with no effect on pro-inflammatory cytokine (TNF-α) production. Pretreating mice with carrageenan once a day before injecting LPS increased the levels of IL-10 by 2.5-fold and reduced TNF-α production by 2-fold compared with control. So, κ/β-carrageenan alone and in combination with LPS enhanced the cellular activity and mobility of peritoneal macrophages by increasing cell adhesion and migration compared with control. LPS activated cells intensively, sometimes resulting in their destruction by necrosis; carrageenan pretreatment reduced the excessive inflammatory cell activation caused by LPS. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1549-1557, 2017.

Plant lectins ConBr and CFL modulate expression toll-like receptors, pro-inflammatory cytokines and reduce the bacterial burden in macrophages infected with Salmonella enterica serovar Typhimurium

BackgroundPlant lectins have long been used in biomedical research as immunomodulators against tumor cells and microbial infections. PurposeTo test the ability of plant lectins ConBr (Canavalia brasiliensis) and CFL (Cratylia argentea) to activate antimicrobial and immunomodulatory activities of murine peritoneal macrophages (pMØ) infected with a virulent strain of Salmonella enterica serovar Typhimurium (STm). MethodsWe incubated pMØ with non-toxic amounts of ConBr and CFL either before (preventive schedule) or after (curative schedule) exposure to STm. ResultsIn uninfected pMØ, ConBr and CFL greatly increased levels of mRNA transcripts for IL-1β, TNF-α and IL-6 and the inducible nitric oxide synthase (iNOs), but not IL-10 and IL-12. Exposure to naïve splenocytes of culture supernatants of pMØ previously stimulated with CFL resulted in expression of IL-12 and IFN-γ. Both preventive and curative treatment schedules significantly reduced the intracellular load of Salmonella. Experiments in infected macrophages exposed to lectins in the preventive schedule showed that mRNA transcripts for IL-6 and TNF-α were increased by CFL, whereas ConBr enhanced IL-12 (subunit p40). In the curative schedule, CFL induced significant expression of IL-12 (p40) whereas ConBr enhanced expression IL-1β and TNF-α genes. The lectin treatments did not influence on iNOs expression in pMØ infected with STm C5 regardless of the treatment schedule. Curative treatments with CFL increased approximately 130-fold expression of TLR-4 whist expression of TLR-9 was increased by treatments with ConBr. ConclusionWe conclude that lectins ConBr and CFL have immunomodulatory properties that are beneficial on control of cells infected by Salmonella.

Immunomodulatory and cellular anti-oxidant activities of caffeic, ferulic, and p-coumaric phenolic acids: a structure–activity relationship study

Many studies have been performed to assess the potential utility of natural products as immunomodulatory agents to enhance host responses and to reduce damage to the human body. To determine whether phenolic compounds (caffeic, ferulic, and p-coumaric acids) have immunomodulatory effects and clarify which types of immune effector cells are stimulated in vitro, we evaluated their effect on splenocyte proliferation and lysosomal enzyme activity. We also investigated the activity of natural killer (NK) cells and cytotoxic T lymphocytes (CTL). In addition, induction of the cellular antioxidant activity in splenocytes, macrophages, and red blood cells was determined by measuring the fluorescence of the DCF product. The study first results indicated that caffeic, ferulic, and p-coumaric acids significantly promote LPS-stimulated splenocyte proliferation, suggesting a potential activation of B cells, and enhanced humoral immune response in hosts treated by the tested natural products. Phenolic acids significantly enhanced the killing activity of isolated NK and CTL cells but had negligible effects on mitogen-induced proliferation of splenic T cells. We showed that caffeic acid enhances lysosomal enzyme activity in murine peritoneal macrophages, suggesting a potential role in activating such cells. Immunomodulatory activity was concomitant with the cellular antioxidant effect in macrophages and splenocytes of caffeic and ferulic acids. We conclude from this study that caffeic, ferulic, and p-coumaric acids exhibited an immunomodulatory effect which could be ascribed, in part, to their cytoprotective effect via their antioxidant capacity. Furthermore, these results suggest that these natural products could be potentially used to modulate immune cell functions in physiological and pathological conditions.

Mycobacterium tuberculosis antigens MPT63 and MPT83 increase phagocytic activity of murine peritoneal macrophages.

Macrophages (MΦ) are the most described and characterized target and host of mycobacteria. Like other cells of innate immunity MΦ have a wide range of receptor molecules which interact with different pathogen associated molecular patterns (PAMPs). Immunodominant proteins MPT63 and MPT83 that are synthesized in abundance by Mycobacterium bovis or Mycobacterium tuberculosis strains could be involved in development of tuberculosis infection. The aim of this study was to search for effects of these mycobacterial antigens on target cells. For this aim full-sized sequences of MPT83 (rMPT83full) and MPT63 antigens were cloned into plasmid pET24a(+). The increase of phagocytic activity of murine peritoneal macrophages was demonstrated, but not of macrophage-like cells from J774 cell line, which were treated by rMPT63 and rMPT83full proteins for 24 h. This effect of such antigens can be considered as a way to facilitate the consumption of mycobacterial cells by macrophages to avoid other effector mechanisms of innate and adaptive immunity.

Investigation of immunomodulatory and anti-inflammatory effects of eriodictyol through its cellular anti-oxidant activity.

Many studies have been performed to assess the potential utility of natural products as immunomodulatory agents to enhance host responses against infection or to ameliorate immune-based pathologies. To determine whether eriodictyol has immunomodulatory effects and clarify which types of immune effector cells are stimulated in vitro, we investigated the stimulatory effect of eriodictyol on spleen cells isolated from BALB/c mice. Eriodictyol significantly stimulated splenocyte proliferation. However, only B lymphocytes (not T lymphocytes) could be stimulated by eriodictyol in a dose-related manner. Studies assessing potential effect of eriodictyol on innate immunity reported that eriodictyol enhanced significantly the killing activity of natural killer (NK) cells, T lymphocytes, and macrophages. We also demonstrated that eriodictyol inhibited nitric oxide (NO) production and lysosomal enzyme activity in murine peritoneal macrophages cultured ex-vivo, suggesting a potential anti-inflammatory effect in situ. Eriodictyol revealed also a cellular anti-oxidant activity in splenocytes and macrophages. Furthermore, eriodictyol increased catalase activity in spleen cells. From this data, it can be concluded that eriodictyol exhibited an immunomodulatory effect that could be ascribed in part to a cytoprotective effect related to its anti-oxidant activity.

Comparative phytochemical profiling and effects of Nerium oleander extracts on the activities of murine peritoneal macrophages

Nerium oleander is a medicinal plant. Apart from its ethnopharmacological uses, pharmacognostic studies have revealed several of its bioactivities. Previously we demonstrated that the phenolic and flavonoid rich extracts of oleander leaf, stem and root possess potent antioxidant and free radical scavenging activities. Moreover, the leaf extract actively modulates the Th1/Th2 cytokine balance and exerts anti-inflammatory activities on murine splenic lymphocytes. Therefore, the present study was designed to evaluate the effect of oleander leaf, stem and root extracts on phagocytosis and the free radical-related activities of murine peritoneal macrophages. In addition, phytochemical profiling was performed using gas chromatography-mass spectrometry (GC-MS). The results demonstrated that the increase in phagocytosis and decrease in myeloperoxidase (MPO) were in the order of leaf&gt;root&gt;stem. The inhibition of cell adhesion, nitric oxide (NO) and elevation of respiratory burst activity was in the order of leaf&gt;stem&gt;root. However, the bioactivities of the leaf extract were much high than those of the stem and root extracts. Phytochemical analysis also revealed the presence of several bioactive constituents in oleander extracts. Therefore, the present study demonstrated that oleander possesses the capacity to modulate macrophage activities and the bioactivities are attributed to the numerous phytochemicals identified in oleander extracts.

Influence of tetrapeptide tuftsin on intracellular pH of mouse peritoneal macrophages

It has been shown by the method of probe microfluorimetry of single cells that tuftsin at concentrations of 0.1 and 1.0 μg/mL, increasing the phagocytic activity of murine peritoneal macrophages, causes biphasic changes in the intracellular pH (pHi) over time. First there was a decrease in pHi, reaching the limit in 5 min of incubation. Then, the pHi value increased until reaching the maximum 30 min after the interaction between the cells and the agent. Afterwards, the observed parameter (pHi) did not vary up to minute 55. It was shown that there was no increase in intracellular pH on the addition of the Na+/H+-exchange blocker ethylisopropylamiloride to the incubation medium in the presence of tuftsin. This fact suggests that the observed increase in pHi during tuftsin treatment in the second phase of cell response is associated with the Na+/H+ exchange system.

The MSHA strain of Pseudomonas aeruginosa (PA-MSHA) inhibits gastric carcinoma progression by inducing M1 macrophage polarization.

Macrophages play crucial roles in promoting tumor development and progression. In the present study, we found that the mannose-sensitive hemagglutination pilus strain of Pseudomonas aeruginosa (PA-MSHA) was efficient in inducing M1 macrophage polarization. PA-MSHA treatment increases expression of M1-related cytokines and promotes activation of murine peritoneal macrophages (MPM). Interestingly, PA-MSHA inhibits cell proliferation and migration and induces the apoptosis of gastric carcinoma cells. These effects of PA-MSHA on M1 polarization were associated with activation of NF-κB expression. Thus, inducing polarization of M1 by PA-MSHA may be one potential strategy for inhibiting gastric carcinoma progression in mice.

Activation effect of Ganoderma lucidum polysaccharides liposomes on murine peritoneal macrophages

The activation of murine peritoneal macrophages by Ganoderma lucidum polysaccharides liposomes (GLPL) was investigated in vitro. After treatment with GLPL, the changes of the nitric oxide (NO) secretion and iNOS (inducible nitric oxide synthase) activity were evaluated. The results showed that NO production and iNOS activity of macrophages were enhanced compared to GLP and BL group. In addition, both the phagocytic activity and levels of cytokines IL-1β, TNF-α and IFN-γ were enhanced in the peritoneal macrophages of mice by stimulation of GLPL. The expression of the major histocompatibility complex class II molecule (MHC II) on the surface of peritoneal macrophages significantly increased. These indicated that GLPL could enhance the activation of peritoneal macrophages and their potential for use as a delivery system of GLP.

녹차 효소 처리 다당의 화학적 특성 및 면역증진 활성

In order to develop new immuno-stimulating ingredients from mature leaves of green tea, crude polysaccharides were isolated from pectinase digests of tea leaves (green tea enzyme digestion, GTE-0), after which their immuno-stimulating activities and chemical properties were examined. GTE-0 mainly contained neutral sugars (54.9%) such as glucose (14.2%), arabinose (12.2%), rhamnose (11.1%), and galacturonic acid (45.1%), which are characteristic of pectic polysaccharides. The anti-complementary activity of GTE-0 was similar to that of polysaccharide K (used as positive control). Number of morphologically activated macrophages was significantly increased in the GTE-0-treated group. GTE-0 significantly augmented H₂O₂ and reactive oxygen species production by murine peritoneal macrophage cells in a dose-dependent manner, whereas production of nitric oxide showed the highest activity at a dose of 100 μg/mL among all tested concentrations. Murine peritoneal macrophages stimulated with GTE-0 showed enhanced production of various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factors-α in a dose-dependent manner. Further, GTE-0 induced higher phagocytic activity in a dose-dependent manner. In ex vivo assay for cytolytic activity of murine peritoneal macrophages, GTE-0-treated group showed significantly higher activity compared to the untreated group at an effector-to-target cell ratio of 20. The above results lead us to conclude that polysaccharides from leaves of green tea have a potent immuno-stimulating effect on murine peritoneal macrophage cells.

In-vitro antimicrobial activity and complement/macrophage stimulating effects of a hot-water extract from mycelium of the oyster mushroom Pleurotus sp.

Abstract The study examined the in vitro antimicrobial and the complement/macrophage stimulating effects of a hot-water extract from mycelium of the oyster mushroom Pleurotus sp. The extract activated the microbial autolytic system of eight strains: seven autolyzing strains with intensity values (IS) ranging from 2.7% in Candida sp. to 36.1% in Saccharomyces cerevisiae, while autolysis was of 1.8% in one non-autolyzing strain (Bacillus cereus). The activation of the alternative pathway of the complement (APC) was dose and time dependent as judged by the lysis of rabbit red blood cells. Two main polysaccharide fractions of the extract significantly bind human immunoglobulin G (IgG) which could result in activation of the classical pathway of the complement (CPC). The extract (5–100 μg/well) enhanced the acid phosphatase activity in murine peritoneal macrophages to 133–184% compared to controls. The findings introduce a novel “bifunctional” approach (antimicrobial-immunomodulatory) to the nutraceutical potential of the Pleurotus hot-water mycelial extract. Industrial relevance At present, between 80% and 85% of all edible-medicinal mushroom products are derived from the fruiting bodies and only 15% are based on extracts from mycelia. The present study suggests that not only Pleurotus mushrooms but also their mycelia may be a good renewable and easily accessible resource for developing functional foods/nutraceuticals or even pharmaceutical agents with antimicrobial and immunomodulatory effects. Additionally, the application of the extract as food bioingredient could represent an innovative strategy for preventing and/or reducing the negative effects of food microbial spoilage. Hence the hot-water extract from mycelia of the oyster mushroom Pleurotus sp. is considerably relevant to the food and pharmaceutical industries.

Effects of Carboxylated Multiwalled Carbon Nanotubes on the Function of Macrophages

Multiwalled carbon nanotubes (MWCNTs) have tremendous potential in many areas of research and applications. Modification of MWCNTs with carboxyl group is one of the widely used strategies to increase their water dispersibility. However, the effect of carboxylation of MWCNTs on their interaction with macrophages remains unclear. The current study compared the impact of pristine MWCNTs (p-MWCNTs) and carboxylic acid functionalized MWCNTs (MWCNTs-COOH) on RAW264.7 cells by looking at the cell viability, phagocytic activity, production of cytokines (IL-1β, IL-10, IL-12, and TNF-α), and intracellular reactive oxygen species (ROS). It was revealed that exposure to either p-MWCNTs or MWCNTs-COOH induced decreased viability of murine macrophage RAW 264.7 cells and moderately elevated phagocytic activity of murine peritoneal macrophages, but no statistical significance was found between the two groups. Increased production of ROS in macrophages was induced after exposure to either p-MWCNTs or MWCNTs-COOH. However, no significantly elevated production of cytokines (IL-1β, IL-10, IL-12, and TNF-α) was observed from RAW 264.7 cells after exposure to the CNTs. Those data suggested that modification with carboxyl group did not exert obvious impact on the interaction of MWCNTs with macrophages.

Effect of Dexamethasone, LPS, or INFγ on the production of TNFα, IL-12, IL-6, IL-10, CXCL8, CXCL10, and CLC3, by bovine macrophages infected in vitro with Mycobacterium avium paratuberculosis.

Effect of Dexamethasone, LPS, or INFγ on the production of TNFα, IL-12, IL-6, IL-10, CXCL8, CXCL10, and CLC3, by bovine macrophages infected in vitro with Mycobacterium avium paratuberculosis.

Immune stimulating and therapeutic potential of tuftsin-incorporated nystatin liposomes against Cryptococcus neoformans in leukopenic BALB/C mice

Cryptococcus neoformans infection is a common fungal infection in persons infected with human immune deficiency virus (HIV) or those with defective cell-mediated immunity. Since treatment of cryptococcal meningitis poses a big challenge, the present study aimed to develop a novel liposomal therapeutic formulation against cryptococcosis. Treatment with tuftsin-incorporated liposomes increased the anti-cryptococcal activity of murine peritoneal macrophages. Prophylactic treatment of mice with tuftsin-incorporated liposomes reduced the dissemination of C. neoformans to brain tissues. Moreover, the co-administration of tuftsin with nystatin liposomes augmented the anti-cryptococcal activity of nystatin, as mice treated with tuftsin-incorporated nystatin liposomes showed the highest survival and least fungal burden in their brain tissues. The results of the present study favour the use of immune-stimulating molecules along with antifungal agents in the treatment of opportunistic fungal infections.

らい菌貪食に伴うマクロファージの抗&lt;i&gt;Mycobacterium intracellulare&lt;/i&gt; 殺菌能の低下

Patients with highly bacillated lepromatous leprosy (LL) essentially lack T cell-mediated immune responses specific to Mycobacterium leprae (ML) antigens, resulting in severely impaired host resistance to leprosy bacilli. Such type of immune unresponsiveness characteristic of LL patients is mainly attributable to markedly depressed T cell ability to activate/expand in response to ML antigens. In this study, we examined profiles of antimycobacterial activity of macrophages, which phagocytized leprosy bacilli, because there is another possibility that, in LL patients, host macrophages in the leprosy lesions are impaired in their antimicrobial activity due to their interaction with infected leprosy bacilli, particularly cellular events through binding with and/or internalization of the pathogens, thereby causing the reduction in host resistance to ML pathogens. The present study indicated the following. First, the anti-M. avium complex activity of murine peritoneal macrophages was significantly reduced when they had phagocytosed heat-killed leprosy bacilli. Second, infection of macrophages with leprosy bacilli did not affect macrophage-mediated suppressor activity against T cell proliferative response to Concanavalin A. These findings indicate that macrophage's intracellular signaling pathways that are up-regulated in response to phagocytosis of leprosy bacilli are linked to the signaling cascades participating in macrophage antimicrobial functions, but not cross-talk with those allowing the expression of macrophage's suppressor activity against T cell functions.

In vitro activation of murine peritoneal macrophages by recombinant YopJ: Production of nitric oxide, proinflammatory cytokines and chemokines

Recently it was reported that 3 μg/ml of recombinant YopJ induced apoptosis in murine peritoneal macrophages in vitro. However, in this study, we report the activation of murine peritoneal macrophages in vitro on treatment with sub-apoptotic dose of recombinant YopJ protein (1 μg/ml). The activation involves enhanced production of nitric oxide (NO), tumor necrosis factor-α (TNF-α), IL-12, and IL-6. Production of NO and IL-6 was found to peak at 24 h of rYopJ treatment, whereas IL-12 and IFN-γ production peaked at 18 h of rYopJ treatment. Increased mRNAs expression of nitric oxide, IL-12, IL-6 and IFN-γ molecules, was also observed in rYopJ-treated macrophages by RT-PCR. rYopJ induced the enhanced activity of protein tyrosine kinases which was inhibited by pharmacological inhibitor genestein, wortmanin and H-7 suggesting the role of tyrosine kinases, PI3K and PKC in the above process. rYopJ also induced increased enhanced production chemokines MIP-1α, MCP-1, and RANTES in macrophages. Significantly, increased expression of TLR-2, TLR-6, MyD 88 and IRAK-1 was also observed by immunoblotting in rYopJ-treated macrophages. rYopJ induced production of NO, TNF-α and IL-6 was significantly inhibited in macrophages pretreated with pharmacological inhibitor wortmanin, genestein and H-7 demonstrating the probable involvement of protein tyrosine kinases in the above process.

Methylglyoxal induced activation of murine peritoneal macrophages and surface markers of T lymphocytes in Sarcoma-180 bearing mice: Involvement of MAP kinase, NF-κβ signal transduction pathway

Methylglyoxal profoundly stimulates host's immune response against tumor cell by producing reactive oxygen intermediates (ROI's) and reactive nitrogen intermediates (RNI's) [Bhattacharyya, N., Pal, A., Patra, S., Haldar, A.K., Roy, S., Ray, M., 2008. Activation of macrophages and lymphocytes by methylglyoxal against tumor cells in the host. Int. Immunophar. 8 (11), 1503–1512]. Present study indicated that methylglyoxal stimulates iNOS activation by p38 MAPK-NF-κβ dependent pathway and ROS production by ERK and JNK activation in sarcoma-180 tumor bearing mice. Proinflammatory cytokines, for macrophage activation, IL-6 and IL-1β were also increased. Production of TLR 4 and TLR 9, which acts through the same signaling pathway, were also upregulated. Hence, concluded that methylglyoxal augmented the IL-6 and IL-1β, expression of TLR 4 and TLR 9 and produced MAPKs, important regulators of ROIs and RNIs. Methylglyoxal treatment also increased M-CSF, an upregulator of macrophage production. CD8 and CD4 molecules, associated with TC and TH cells respectively, were also increased. Overall methylglyoxal treatment is important for enhancement of macrophages and lymphocyte activation or immunomodulation against sarcoma-180 tumor.

Effect of interferon-gamma priming on the activation of murine peritoneal macrophages to tumouricidal state by cisplatin, IL-1, and tumour necrosis factor (TNF): production of IL-1 and TNF

The effect of interferon-gamma (IFN-gamma) priming of murine peritoneal macrophages on the activation to tumouricidal state by cisplatin, lipopolysaccharide (LPS)-IL-1 and TNF was investigated. Cisplatin-, LPS-, IL-1- or TNF-treated IFN-gamma-primed macrophages showed significantly enhanced tumouricidal activity and binding to tumour cells, compared with unprimed treated or untreated macrophages. Macrophages treated with cisplatin, LPS, IL-1 and TNF produced released and membrane-associated IL-1 and TNF activity which was significantly enhanced after priming with IFN-gamma. These observations suggest the use of IFN-gamma along with these biological response modifiers in designing immunotherapeutic protocols for treatment of malignancy.