Activity Of Peritoneal Macrophages Research Articles

Crotoxin modulates metabolism and secretory activity of peritoneal macrophages from Walker 256 tumor-bearing rats

Crotoxin (CTX), the major toxin of Crotalus durissus terrificus snake venom, induces an inhibitory effect on tumor development and modulates the functions of macrophages (MØs), which play a key role as a defense mechanism against tumor growth. In early tumor progression stage, MØs are avidly phagocytic (inflammatory cell), releasing reactive nitrogen intermediates-RNI/ROI and cytokines TNF-α, IL-1β, and IL-6. However, when the tumor has been developed, tumor-associated MØ (angiogenic cell) presents a decrease in the mentioned activities. We reported that CTX stimulates H2O2 release, NO production and secretion of cytokines by peritoneal MØs obtained from non-tumor-bearing rats. Considering that the mentioned mediators control tumor growth, it is mandatory to investigate whether CTX stimulates the production of these mediators by MØs obtained from tumor-bearing animals. The aim of this work was then to evaluate the CTX effect on metabolism and functions of peritoneal MØs obtained from Walker 256 tumor-bearing rats. For this purpose, male Wistar rats were subcutaneously inoculated in the right flank with 1 mL sterile suspension of 2 × 107 Walker 256 tumor cells. CTX (18 μg per animal) was subcutaneously administered in two protocols: a) on the 1st day of tumor cell injection and b) on the 4th day of tumor cell inoculation. In both protocols, MØs were obtaining on the 14th day of tumor cell inoculation to evaluate the release of H2O2, NO, and pro-inflammatory cytokines (IL-1β, TNFα, and IL-6); maximal activity of hexokinase, glucose-6-phosphate dehydrogenase, citrate synthase, and 14CO2 production from [U–14C]-glucose and [U–14C]-glutamine. The treatment with CTX stimulated the release of NO, H2O2, and cytokines, and glucose and glutamine metabolism. Metabolic and functional changes induced by CTX were accompanied by a decrease of tumor growth as indicated by tumor fresh weight and diameter. These results indicate CTX not only as a scientific tool to investigate changes in metabolism and functions of peritoneal MØs but also for a better understanding of the mechanisms involved in tumor growth.

CD146 Defines a Mesenchymal Stromal Cell Subpopulation with Enhanced Suppressive Properties.

Mesenchymal stromal cells (MSCs) are largely known for their immune-suppressive capacity, hence, their common use in the control of unwanted inflammation. However, novel concepts related to their biology, combined with the urgent need to identify MSC subpopulations with enhanced suppressive properties, drive the search for isolation protocols optimized for clinical applications. We show, in this study, that MSCs expressing high CD146 levels exhibit altered surface expression profiles of CD44 and secrete elevated levels of interleukin (IL)-6, amongst other factors. In addition, CD146hi MSCs surpass the polyclonal parental populations in inhibiting alloreactive T cells in vitro, in both a soluble- and cell-contact-dependent manner. Despite the lack of CD146hi MSC-mediated activation of peritoneal macrophages to release the suppressive factor IL-10 in vitro, their administration in animals with graft-versus-host disease alleviates inflammation and leads to 40% survival rate up to 7 weeks post-transplantation. This pronounced inhibitory property is driven by CD146-mediated in situ efferocytosis by myeloid cells. Altogether, this study provides the impetus to adopt an isolation protocol for MSCs based on a CD146 expression profile before their therapeutic use and suggests a major role played by CD146 as a novel “eat-me” signal, capable of enhancing MSC uptake by competent phagocytes.

Low-dose mycophenolate mofetil improves survival in a murine model of Staphylococcus aureus sepsis by increasing bacterial clearance and phagocyte function.

Regulators of TLRs signaling pathways play an important role in the control of the pro-inflammatory response that contributes to sepsis-induced tissue injury. Mycophenolate mofetil, an immunosuppressive drug inhibiting lymphocyte proliferation, has been reported to be a regulator of TLRs signaling pathways. Whether MMF used at infra-immunosuppressive doses has an impact on survival and on innate immune response in sepsis is unknown.C57BL/6J mice were infected intraperitoneally with 108 CFU Staphylococcus aureus, and treated or not with low-dose of MMF (20mg/kg/day during 4 days). Survival rate and bacterial clearance were compared. Cytokine levels, quantitative and qualitative cellular responses were assessed. S. aureus – infected mice treated with MMF exhibited improved survival compared to non-treated ones (48% vs 10%, p<0.001). With the dose used for all experiments, MMF did not show any effect on lymphocyte proliferation. MMF treatment also improved local and systemic bacterial clearance, improved phagocytosis activity of peritoneal macrophages resulting in decreased inflammatory cytokines secretion. MMF-treated mice showed enhanced activation of NF-κB seemed with a suspected TLR4-dependent mechanism. These results suggest that infra-immunosuppressive doses of MMF improve host defense during S. aureus sepsis and protects infected mice from fatal outcome by regulating innate immune responses. The signaling pathways involved could be TLR4-dependent. This work brings new perspectives in pathogenesis and therapeutic approaches of severe infections.

Functional Phenotypes of Peritoneal Macrophages Upon AMD3100 Treatment During Colitis-Associated Tumorigenesis.

CXCL12 and its receptor CXCR4 are independent prognostic factors in colorectal cancer. AMD3100 is the most frequently used FDA-approved antagonist that targets the CXCL12-CXCR4 axis in clinical trials. We aimed to explore the role of AMD3100 and its effect on peritoneal macrophages' functional phenotypes during colitis-associated tumorigenesis. We treated AMD3100 in a colitis-associated colon cancer mouse model and evaluated its effect on tumorigenesis. The phagocytosis activities of peritoneal macrophages were measured by flow cytometry. The proportions of macrophages and M1/M2 subpopulations were investigated by flow cytometry, ELISA, and immunochemistry. Serum levels of pro-inflammatory and anti-inflammatory cytokines were measured by LEGENDplex™ kits. Transwell assay and qRT-PCR were performed to investigate the direct effect of CXCL12 on macrophages in vitro. We demonstrated that AMD3100 treatment reduced the inflammatory damages in the colonic mucosal and ameliorated tumor development in experimental mice. We found that the phagocytosis activities of peritoneal macrophages fluctuated during colitis-associated tumorigenesis. The proportions of peritoneal macrophages and M1/M2 subpopulations, together with their metabolite and cytokines, changed dynamically in the process. Moreover, AMD3100 regulated the functional phenotypes of macrophages, including reducing the recruiting activity, promoting polarization to the M1 subpopulation, and reducing IL-12 and IL-23 levels in serum. Our study contributes to understanding dynamic changes of peritoneal macrophages upon AMD3100 treatment during tumorigenesis and sheds light on the potential therapeutic target of AMD3100 and peritoneal macrophages against colitis-associated colon cancer.

Abstract 421: Hematopoietic Deficiency In NLRP3 And AIM2 Inflammasomes Or Gasdermin D Fails To Prevent Atherosclerosis Progression In Mouse Models Of Diabetes

Diabetes accelerates atherosclerosis progression in part by increasing lesional necrotic core expansion, but the underlying mechanisms remain elusive. Previous studies have shown that systemic inhibition of the NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome prevents atherosclerosis progression in a mouse model of diabetes. Gasdermin D (GSDMD), which acts downstream of inflammasome activation, contributes to pyroptosis and IL-1β and IL-18 release. To explore if hematopoietic inflammasome activation mediates diabetes-related necrotic core expansion, we first generated hematopoietic GSDMD-deficient chimeras in an LDL receptor-deficient ( Ldlr -/- ) virally-induced mouse model of type 1 diabetes (T1D)-accelerated atherosclerosis. Mice with T1D demonstrated elevated plasma glucose, triglycerides, cholesterol, and plasma IL-18 (non-diabetes [ND]=180.4 pg/ml, diabetes [D]=316.6, N=17-18 per group). However, diabetic mice with hematopoietic GSDMD-deficiency had no reduction in plasma IL-18 (D-KO=437.5 pg/ml, p=0.29, N=17). Moreover, we found little evidence of inflammasome activation in peritoneal and splenic macrophages from diabetic mice. A 4-week duration of diabetes significantly increased necrotic core area measured at three different levels in the aortic sinus (p&lt;0.01, N=21-24 per group) in pre-existing lesions without increasing lesion area. Hematopoietic GSDMD-deficiency did not affect the necrotic core area. To further interrogate the relationship between hematopoietic inflammasome activation and diabetes-related necrotic core expansion, we generated Ldlr -/- hematopoietic NLRP3 and absent in melanoma 2 (AIM2) inflammasome double knockout (DKO) chimeras in a streptozotocin mouse model of diabetes. Mice with diabetes demonstrated hyperglycemia and hypercholesterolemia. However, diabetic hematopoietic DKO chimeras did not exhibit reduced aortic sinus necrotic core area, as compared with diabetic wildtype controls. Our findings suggest that the increased plasma IL-18 levels in diabetic mice are not dependent on hematopoietic GSDMD, and that diabetes-mediated necrotic core expansion is independent of hematopoietic NLRP3 and AIM2 inflammasome and pyroptosis pathways.

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.

OI inhibits development of ovarian cancer by blocking crosstalk between cancer cells and macrophages via HIF-1α pathway

Intraperitoneal implantation of ovarian cancer (OC) decreases the survival rate in clinical practice. However, there are still great deficiencies in the therapeutic strategies of inhibiting the metastasis of OC. Here, we showed that 4-octyl itaconate (OI, a cell-permeable itaconate derivative) treatment didn't influence the development of OC in vitro. Instead, OI treatment repressed the activation of peritoneal macrophages and downregulated the expression of proinflammatory factors in mice bearing OC. Besides, OI inhibited the angiogenesis of OC tissues by downregulating HIF-1α of OC that was induced by proinflammatory factors from activated macrophages. In conclusion, our findings demonstrate that OI suppresses metastasis of ovarian cancer by blocking crosstalk between cancer cells and macrophages via HIF-1α pathway, providing a potential therapeutic strategy for metastasis of OC.

Ligstroside aglycon, an extra virgin olive oil secoiridoid, prevents inflammation by regulation of MAPKs, JAK/STAT, NF-κB, Nrf2/HO-1, and NLRP3 inflammasome signaling pathways in LPS-stimulated murine peritoneal macrophages.

Ligstroside aglycon (LA) is one of the main polyphenols in extra virgin olive oil (EVOO); nevertheless, it is scarcely investigated. The aim of this study was to evaluate the immunomodulatory and anti-inflammatory effects of LA on lipopolysaccharide (LPS)-stimulated murine peritoneal macrophages, as well as the potential signaling pathways involved. Isolated macrophages were treated with LA (50, 25, and 12.5 μM) in the presence or absence of LPS (5 μg ml-1) for 18 h. Cell viability was determined using the sulforhodamine B (SRB) assay. Nitric oxide (NO) and pro-inflammatory cytokine production was analyzed by the Griess method and enzyme-linked immunosorbent assay (ELISA), respectively. Protein expression of pro-inflammatory markers and signaling pathways were evaluated by western blot analysis. LA showed significant antioxidant and anti-inflammatory effects through decreasing oxidative stress markers such as NO production, inducible nitric oxide synthase (iNOS) and NADPH oxidase-1 (NOX-1) protein expression. Besides, LA was able to reduce pro-inflammatory cytokine levels and modulate cyclo-oxygenase-2 (COX-2), and microsomal prostaglandin E synthase-1 (mPGEs-1) protein overexpression. The mechanisms underlying these protective effects could be related via activation of nuclear factor (erythroid-derived 2)-like (Nrf2)/heme oxygenase 1 (HO-1) and inhibition of nuclear factor kappa-B (NF-κB), mitogen-activated protein kinases (MAPKs), and Janus kinase/signal transducer and activation of transcription (JAK2/STAT3) signaling pathways. In addition, LA inhibited non-canonical and canonical activation of a nucleotide-binding (NOD)-like receptor (NLRP3) inflammasome. We conclude that LA showed significant antioxidant and anti-inflammatory activities in LPS-stimulated murine peritoneal macrophages. However, further in vivo studies are warranted to further investigate the bioactivity of this interesting compound that might be a promising natural agent for the treatment of immune-inflammatory diseases.

The effect of water-soluble pristine C60 fullerene on 6-OHDA-induced Parkinson’s disease in rats

Oxidative stress is thought to be one of the mechanisms that leads to the dysfunction and degeneration of dopaminergic neurons in Parkinson’s disease pathogenesis and presumed to be underway during the prodromal phase. Therefore, therapy, which is effective against pre-motor symptoms, might be effective in preventing or delaying the development and progression of Parkinson’s disease. The aim of our study was to investigate the therapeutic efficiency of pristine C60 fullerene aqueous solution (C60FAS) during Parkinson’s disease in rats. The unilateral dopamine deficiency was induced in male Wistar rats (220–250 g) by stereotaxic microinjection of neurotoxin 6-hydroxydopamine (6-OHDA, 12 μg). C60FAS was injected to rats intraperitoneally daily for 10 days (0.65 mg/kg per day). The percentage of destroyed dopaminergic neurons was determined by the apomorphine test and by IHC staining of tyrosine hydroxylase-positive neurons in substantia nigra. We evaluated the rat body weight, the water and food intake, Open Field behavioural test, the level of biochemical antioxidant system, the activity of peritoneal macrophages. Levels of spontaneous and carbachol-stimulated colon motility were estimated by ballonographic method in vivo. C60FAS showed a positive tendency to increase the number of tyrosine hydroxylase-positive cells in the midbrain, which was associated with more profound improvement in apomorphine-rotation behaviour and slight relief of the anxiety level in Open Field test. Furthermore, C60FAS treatment increased the index of stimulated distal colon motor activity while it did not have a significant effect on water content in feces and total gastrointestinal transit time. C60FAS treatment did not affect water intake behaviour or body weight changes while it induced an increase of glutathione level and decrease activity of glutathione peroxidase in the brain as well as an increase in activity of peritoneal macrophages in 6-OHDA-Parkinson’s disease rats. These findings confirmed the potential therapeutic effectiveness of water-soluble pristine C60 fullerene in Parkinson’s disease pathogenesis, though there is ground for caution because of its systemic mild toxic effect.

Comparison of the effects of co-transplantation of bone marrow hematopoietic stem cells and thymic multipotent stromal cells on the immune system of mice depending on methods

Physical interaction of multipotent stromal cells (MSCs) and hematopoietic stem cells (HSCs) is a modern approach to effective and focused changes in the properties of HSCs. Resulting of those contact interaction is significant activation of cells with following immune system restoration.&#x0D; The purpose of the study is to investigate the effect of co-transplantation of bone marrow hematopoietic stem cells (HSCs) and thymic multipotent stromal cells (MSCs) separately and as a union of cells on regeneration of the murine immune system, damaged by cyclophosphamide.&#x0D; MSCs were obtained from thymuses of C57BL mice using explant technique. Bone marrow cells (BMCs) were obtained by flushing out the femur with a nutrient medium. BMCs were cocultivated for 2 hours on the monolayer of thymus-derived MSCs. The immune deficiency of mice was modelled by the treatment with cyclophosphamide (CP). After that, the cells were co-transplanted in two methods (separately into different the retroorbital sinus and as a union after co-cultivation) and the parameters of the immune system were evaluated. It was shown, that separate co-transplantation of BMCs and thymus-derived MSCs is associated with the restoration of the number of bone marrow cells, thymus, spleen and lymph nodes with an increase in the proliferation index of lymph node cells by 1.4 times compared to control. It normalized the previous reduced concentration of hemoglobin and hematocrit in the blood. Co-transplantation had a suppressive effect on the blast transformation reaction, induced by phytohemagglutinin, by 4.3 times, but showed a stimulating effect on DTHR response by 1.6 times compared to control.&#x0D; Co-transplantation of the union of BMCs and MSCs is associated with the restoration of the number of bone marrow cells, spleen and lymph nodes. The level of spontaneous apoptosis of lymph node cells significantly increased by 3.3 times compared to control. It had not effect on hematological parameters, but is activated to impact the immune system. Thus, as a result of cells union administration showed normalization of the bactericidal activity of peritoneal macrophages, unlike the separate co-transplantation. This cells graft had a suppressive effect on the number of antibody-producing cells in the spleen by 4.2 times compared to control.&#x0D; Previous co-cultivation and contact interaction of cells change the properties of cell graft. The effect of co-transplantation of BMCs and thymic MSCs is not a simple additive effect of cells. It is acquiring the features typical to certain cell types, and the expression of new characteristics. We assume this phenomenon as a result development of complex cells cooperative processes in vivo and in vitro

Cytotoxicity and Anti-Inflammatory Effects of Polyherbal Formulations, Joint Pain Spl and Rumalaya Forte on Lipopolysaccharide Induced Inflammation in IC-21 Macrophages.

To test the effectiveness of marketed polyherbal formulations on lipopolysaccharide-induced inflammatory conditions in macrophages. Usage of herbal compounds among patients suffering from arthritis and cancer is increasing every year. Many anti-inflammatory herbal products available in the market should be screened thoroughly for their possible mechanism of action. Joint Pain Spl (JPS) is a polyherbal dietary food supplement composed of 13 herbal plants, and Rumalaya Forte (RF) is a polyherbal formulation comprising of 6 herbal plants. These were tested for their cytotoxicity, as well as antioxidant and anti-inflammatory activities in LPS treated IC-21 peritoneal macrophages. Commercially available JPS and RF powder was used to prepare the extract. The aqueous and methanol extracts were quantified for the presence of phenolic and flavonoid compounds and confirmed with HPLC. In vitro DPPH free scavenging activity was performed. Cytotoxicity was tested by MTT assay. Anti-inflammatory activity was tested using lipopolysaccharide-stimulated IC-21 peritoneal macrophage cells. The phytochemical screening showed the presence of phenolic and flavonoid compounds in JPS and RF. The aqueous and methanol extracts of JPS and RF possesses significant DPPH free radical scavenging activity. MTT assay revealed that 90.64% (aqueous extract) and 92.21% (methanol extract) of exposed macrophages are viable even after 24h exposure of maximal tested concentrations of herbal formulations. Pre-treatment of JPS and RF on LPS induced IC-21 macrophages showed a reduction in nitric oxide production (maximal 79.95%) and a high level of superoxide anion scavenging activity (maximal 82.5%) over control. The two tested polyherbal formulations, such as JPS and RF possess anti-inflammatory activity by modulating free radical generation in IC-21 macrophages. Thus the presence of the phenolic and flavonoid compounds may contribute to the antioxidant activity.

Ethanolic Extract of Black Cumin Seed Reduced Radical Reactive from Dimethylbenzanthracene Compounds

Dimethylbenzantracene is a compound that is radical reactive, genotoxic, and immunotoxic. Black cumin seeds have antioxidative properties. The study aimed to determine the effect of the black cumin seed ethanolic extract as an antioxidant by observing the secretion activity of ROI and NO peritoneal macrophages in Sprague Dawley (SD) rats induced by DMBA. This controlled experiment used 30 male SD rats. The test animals were randomly divided into six groups. Group I was given standard food and drink. Group II was given DMBA. Groups III, IV, and V were given EEBCS doses of 5, 25, and 125 mg orally for seven weeks, respectively. Group VI was given Imboost. From week 3 to week seven, all test animals were induced 10x20 DMBA, 2x per week.At week 16, all test animals were sacrificed and isolated and cultured peritoneal macrophage cells. The activity of ROI secretion and NO macrophage secretion was tested. Data on ROI secretion and NO levels were tested for ANOVA, followed by LSD test with a confidence level of 95%. The results showed that DMBA induction increased ROI secretion by macrophages and decreased NO.EEBCS secretion decreased ROI secretion and grew NO. The DMBA group had the lowest NO levels. This study concluded that EEBCS at doses 5 and 125 reduced the flow of ROI peritoneal macrophages in SD rats induced by DMBA but reduced NO levels.

Ongoing Exposure to Peritoneal Dialysis Fluid Alters Resident Peritoneal Macrophage Phenotype and Activation Propensity.

Peritoneal dialysis (PD) is a more continuous alternative to haemodialysis, for patients with chronic kidney disease, with considerable initial benefits for survival, patient independence and healthcare costs. However, long-term PD is associated with significant pathology, negating the positive effects over haemodialysis. Importantly, peritonitis and activation of macrophages is closely associated with disease progression and treatment failure. However, recent advances in macrophage biology suggest opposite functions for macrophages of different cellular origins. While monocyte-derived macrophages promote disease progression in some models of fibrosis, tissue resident macrophages have rather been associated with protective roles. Thus, we aimed to identify the relative contribution of tissue resident macrophages to PD induced inflammation in mice. Unexpectedly, we found an incremental loss of homeostatic characteristics, anti-inflammatory and efferocytic functionality in peritoneal resident macrophages, accompanied by enhanced inflammatory responses to external stimuli. Moreover, presence of glucose degradation products within the dialysis fluid led to markedly enhanced inflammation and almost complete disappearance of tissue resident cells. Thus, alterations in tissue resident macrophages may render long-term PD patients sensitive to developing peritonitis and consequently fibrosis/sclerosis.

Experimental study on disorders in the activity of macrophages in the acute period of different severities of cerebral stroke

BACKGROUND: The development of immunosuppression in the acute phase of cerebral stroke may lead to infectious and inflammatory complications. However, data on the functional activity of macrophages are insufficient to understand the pathogenesis of stroke.&#x0D; AIM: This study aimed to investigate the functional activity of macrophages in the acute period of experimental cerebral stroke (ECS) of different severities.&#x0D; MATERIALS AND METHODS: In an experimental study on 45 Wistar rats, the functional and spontaneous metabolic activity of macrophages in the spleen and abdominal cavity and the induced metabolic activity of peritoneal macrophages were evaluated through the reduction of nitro blue tetrazolium via spectrophotometry. The spontaneous and induced adhesive activity and the inhibition of the modified adhesive activity of peritoneal macrophages were examined via crystal violet staining by using a colorimetric method. ECS was modeled by the rotational movements of mandrel-knife inserted into the region of capsula interna (c.i.) on the left by guiding a needlecannula with the subsequent damage of vessels and the formation of mild (local incision of vessels in the region of c.i.), moderate (additional introduction of autologous blood in the area of c.i.), and severe (cutting of vessels from c.i. to pia mater with the subsequent introduction of autologous blood into the damaged area) hemorrhage. Data were evaluated in 72 h after different severities of ECS were modeled.&#x0D; RESULTS: In 72 h after ECS with different severities was modeled, the functional and spontaneous metabolic activity of splenic macrophages significantly decreased compared with those in the control group. The functional, spontaneous, and induced metabolic activity of peritoneal macrophages significantly reduced compared with those in the control group. The adhesive activity of peritoneal macrophages significantly increased compared with that of the control group. These disorders exacerbated in animal models with a more severe degree of stroke.&#x0D; CONCLUSION: In the acute period of ECS, the functional activity of macrophages decreased, whereas their adhesive activity increased. These phenomena enhanced in more severe models of stroke.

Modulating effects of the synergistic combination of extracts of herbal drugs on cyclophosphamide-induced immunosuppressed mice.

ObjectiveTaking leads from the available research, we aimed to develop a synergy-based herbal combination of Tinospora cordifolia (TC), Phyllanthus emblica (PE), and Piper nigrum (PN). Also, evaluating their synergistic effect on CP-induced immunosuppression in mice model and exploring the possible mechanisms involved in reversing the damage.MethodologyThe immunomodulatory activity of combination, of TC stem, PE fruits, and PN dried fruits, was determined by in vitro assays (splenocyte proliferation and pinocytic activity of peritoneal macrophages of mice) and in vivo study using CP-induced immunosuppression model in Swiss Albino mice. The ratio was optimized for combining three by in vitro MTT assay. The combination was further evaluated for anti-oxidant activity by DPPH scavenging method and quantified for its bioactive metabolites by HPTLC. Serum collected on day 0, 4, 7 and 14 was employed for estimation of haematogram (haematocrit, TLC, DLC, and haemoglobin, etc) and immune parameters (IL-10, IL-6 and TNF-α) by ELISA.ResultsThe study demonstrated, that combination of herbal extracts at an intermediate dose could inhibit the proliferation of spleen cells and peritoneal macrophages (P ≤ 0.0001) and induce suppression of pro-inflammatory mediators, and also certified that combination exerts synergized effects. The results showed that the combination possess potential antioxidant activity by DPPH scavenging method (IC50-113.5 µg/ml). It was identified that combination significantly (P ≤ 0.0001) improved the immune markers, haematogram parameters, and histological parameters, with maximum protection offered by an intermediate dose.ConclusionThe results suggested that present combination could be further explored clinically as potent synergy-based therapeutic approach for immune modulation.

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&gt; 0.05). Cellularity of thymus was significantly increased by the extended-release form of Afobazol in dose of 12 mg/kg (p&lt; 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&lt; 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 &gt; 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&gt; 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.

ИЗМЕНЕНИЯ ИММУНОЛОГИЧЕСКОГО ОТВЕТА В РЕАКЦИИ НСТ С ПЕРИТОНЕАЛЬНЫМИ МАКРОФАГАМИ ПОД ДЕЙСТВИЕМ ШИРОКОПОЛОСНОГО ЭЛЕКТРОМАГНИТНОГО ИЗЛУЧЕНИЯ В ЭКСПЕРИМЕНТЕ

Summary The article presents the results of research on the study of the effect of broadband electromagnetic fields on a macroorganism in an experiment. Electromagnetic fields with different characteristics can affect the systems of living organisms in different ways, including the immune system, which is sensitive to electromagnetic influences. Broadband electromagnetic fields have an immunostimulating effect, as evidenced by the increase in the enzymatic activity of peritoneal macrophages in reaction with nitro blue tetrazolium.

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.

Absence of TNF Leads to Alternative Activation in Peritoneal Macrophages in Experimental Listeria Monocytogenes Infection

ABSTRACT Macrophages are crucial effectors of innate immunity against the pathogenic bacterium Listeria monocytogenes. The pro-inflammatory cytokine tumour necrosis factor-α (TNF) has been shown to be crucial for resistance to L. monocytogenes and mice deficient in TNF signalling succumb quickly after infection. However, the mechanisms underlying TNF-mediated defence against L. monocytogenes infection have not been completely elucidated. Here, we demonstrate that TNF concurrently functions to support a pro-inflammatory M1 phenotype while actively blocking macrophage polarization to the M2 phenotype. Compared to WT mice, peritoneal macrophages in TNF-deficient mice inoculated with L. monocytogenes respond with M2 polarization by upregulating Arg1. Consistently, TNF blockade in vitro resulted in M2 polarization in peritoneal macrophages during L. monocytogenes infection. Additionally, TNF promotes the transition from M2 to M1 polarization in peritoneal macrophages. Further investigation of peritoneal macrophage polarization suggested the NF-κB pathway is involved in the TNF-dependent M2 to M1 shift. Conversely, treatment of peritoneal macrophage with a PPARγ agonist blunted the expression of M1 genes induced by TNF and reduced NF-κB signalling pathway activation. Competing signalling mechanisms therefore play an essential role in the ability of peritoneal macrophage to resolve L. monocytogenes infections with TNF playing an essential role in driving M1 polarization. Abbreviations: LPM: large peritoneal macrophage; SPM: small peritoneal macrophage; LLO: listeriolysin O; iNOS: inducible nitric oxide synthase; DCs: dendritic cells