Macrophage Phagocytosis Activity Research Articles

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.

The Activities of Pegagan Embun (Hydrocotyle sibthorpioides Lam.) on TNF-α, Macrophages and Leukocytes Male White Mice Exposed by H5N1 Virus Antigens

Introduction: Pegagan embun (Hydrocotyle sibthorpioides Lam.) has been known to have immunostimulatory activity, it can increase the activity and capacity of mice macrophage phagocytosis at optimum dose of 200 mg/kgbw. Aim: This study aims to determine the activity of the ethanol extract of pegagan embun on TNF-α levels, total macrophages, total leukocytes, and percentage of leukocytes types. Methods: Mice were divided into four groups, one group as control was given 0.5% Na CMC suspension, three groups were given pegagan embun extract at a dose of 10 mg/kgbw, 50 mg/kgbw and 200 mg/kgbw orally for 7 days. On the 8th day, all mice were induced with 0.3 mL H5N1 vaccine subcutaneously, then left for 24 hours. After 24 hours, the mice were sacrificed and then TNF-α levels, total macrophages, total leukocytes, and percentage of leukocytes types were determined. The research data were analyzed using one-way ANOVA (significance was taken at p<0.05) and continued with the Duncan test. Results: The results showed that pegagan embun extract significantly reduce TNF-α levels and total macrophages, also significantly increase total leukocytes and percentage of leukocyte types in mice (p<0.05). Significant increases and decreases occurred on average at doses of 50 mg/kgbw and 200 mg/kgbw. Conclusion: Based on these results, it can be concluded that pegagan embun extract have immunomodulatory activity.

Identification and characterization of immunomodulatory peptides from pepsin\u2013soy protein hydrolysates

To obtain immunomodulatory peptides from isolated soy protein (ISP), pepsin was selected to prepare hydrolysates and 4-h treatment (Pepsin-ISPH4h) showed the highest yield and immunomodulatory activities. The Pepsin-ISPH4h was sequentially fractionated by 30, 10 and 1-kDa molecular weight cut-off (MWCO) membranes, in which 1-kDa MWCO permeate (1P) exhibited the most significant enhancement of phagocytosis activity without causing excessive inflammation as compared with Pepsin-ISPH4h. To further purify and enhance the immunomodulatory activity, 1P was distinct by high-performance liquid chromatography equipped with a reverse-phase column and in vivo immunomodulatory activity of fractions was examined in mice. Fraction 1 (F1) significantly elevated phagocytosis activity of mice spleen macrophages and neutrophils. However, increase of phagocytosis activity did not result from the induction of macrophages M1 or M2 polarization. The immunomodulatory peptide sequence, EKPQQQSSRRGS, from F1 was identified by LC–MS/MS. Phagocytosis activity and macrophage M1 polarization were elevated by synthetic peptide treatment. Hence, our results indicated that isolated soy protein hydrolysates prepared by pepsin could provide a source of peptides with immunomodulatory effects.Graphical

Assessment of the immune responses of macrophages in selective breeding pangasius families (Pangasianodon hypophthalmus) resistant to enteric septicemia of catfish (ESC)

This study aimed to evaluate the non-specific immune responses of macrophages in two selective breeding pangasius family groups: high disease-resistant (A) and low disease-resistant (B) to Enteric Septicemia of Catfish (ESC) caused by Edwardsiella ictaluri. Fish were sampled at five different time points after infection with the bacteria E. ictaluri including before the infection process, 24hpi, 48hpi, 264hpi and 312hpi. There were 372 samples including 192 samples of fish in group A and 180 samples in group B. All of them were used to analyze the amount of melano-macrophage centers in the tissues of liver, kidney and spleen; phagocytosis activity (PA) and phagocytosis index (PI) of the anterior kidney macrophages. Results showed that (1) the number of melano-macrophages of group A fish was higher than that of group B at survey points during the challenge and the difference was significant at 48hpi; (2) PA and PI of group A were higher than that of group B in the period from 48 to 264hpi, specifically, the PI of group A was higher than that of group B, which was statistically significant at 48hpi. The results of this study showed that the differences in non-specific immune responses through melano-macrophages can improve the survival rate and life span of the high disease-resistant family compared to the low disease-resistant family after the challenge with Edwardsiella ictaluri.

Crocodylus siamensis Liver Extracts Exhibit Antimicrobial Activity and Induce Phagocytic Activity in Macrophage Cells

Antimicrobial peptides are documented as having a crucial role in innate immune response. Many reports have studied antimicrobial peptides from the crocodile. Thus, the antimicrobial activity of liver protein extract from Crocodylus siamensis was examined. Our results show that P50 and P80, partially purified crocodile liver protein extract at 45 mg/ml, can inhibit gram-positive bacteria with the highest percentages of inhibition at 48.22 % and 37.54 % respectively, while gram-negative bacteria could be inhibited at 47.31 % and 35.32 %, respectively. However, only P80 was selected for further phagocytosis activity investigation since it showed low cytotoxicity in the macrophage cell line. The results show that P80 can kill bacteria by an intracellular killing process in phagocytic cells of macrophages in a dose-dependent manner. Moreover, the internalization of phagocytosis activity of macrophages was assayed using fluorescein isothiocyanate-labeled Escherichia coli and flow cytometry. The results reveal that P80 can degrade bacteria by inducing phagosome formation in a time and dose-dependent manner (200-800 μg/ml). In these findings, the liver protein extract of Crocodylus siamensis had an enhancing effect on macrophage RAW 264.7 cells in the phagocytosis process.

Immunomodulatory Properties of Polysaccharides from the Coral Pseudopterogorgia americana in Macrophages.

Polysaccharides from marine organisms produce an important regulatory effect on the mammalian immune system. In this study, the immunomodulatory properties of a polysaccharide that was isolated from the coral Pseudopterogorgia americana (PPA) were investigated. PPA increased the expression levels of tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2), but not inducible nitric oxide synthase and nitric oxide, in macrophages. A mechanistic study revealed that PPA activated macrophages through the toll-like receptor-4 and induced the generation of reactive oxygen species (ROS), increased the phosphorylation levels of protein kinase C (PKC)-α, PKC-δ and mitogen-activated protein kinases (MAPK), and activated NF-κB. The inhibition of ROS and knockdown of PKC-α reduced PPA-mediated TNF-α and IL-6 expression; however, the knockdown of PKC-δ significantly increased PPA-mediated TNF-α expression. In addition, the inhibition of c-Jun N-terminal kinase-1/2 and NF-κB reduced PPA-mediated TNF-α, IL-6 and COX-2 expression. Furthermore, the inhibition of ROS, MAPK and PKC-α/δ reduced PPA-mediated NF-κB activation, indicating that ROS, MAPK and PKC-α/δ function as upstream signals of NF-κB. Finally, PPA treatment decreased the phagocytosis activity of macrophages and reduced cytokine expression in bacteria-infected macrophages. Taken together, our current findings suggest that PPA can potentially play a role in the development of immune modulators in the future.

Robust polymeric scaffold from 3D soft confinement self-assembly of polycondensation aromatic polymer

• A homo-polyarylene ether nitrile (homo-PEN) was synthesized via polycondensation. • The homo-PEN was converted to microparticles via emulsion confinement self-assembly. • The obtained microparticles can be hydrolyzed to present surface carboxyl groups. • Protein A was modified on the carboxylated microparticles for antibody separation. • QD embedded hybrid microparticles were used for assessment of phagocytosis activity. Polymeric microparticles obtained via the emulsion confinement self-assembly of block copolymers have been used as the effective scaffold for construction of bioactive agents for various biomedicine applications, however the used block copolymers are normally synthesize via the controlled/living polymerization that involves tedious workload. Herein, we have discovered that a homo-polyarylene ether nitrile (abbreviated as homo-PEN), obtained via the industrially scalable step-growth nucleophilic substitution polycondensation, can be transformed into sub-micrometer sized polymeric microparticles via emulsion confined self-assembly. We found that the size of obtained microparticles was dependent on the molecular weight of homo-PEN or added ethanol content during self-assembly. More interestingly, the surface cyano of as-obtained polymeric microparticles can be partially converted into carboxyl groups after hydrolysis in alkaline aqueous solution without any morphology destruction. Consequently, the protein A was further covalently grafted on the carboxylated PEN microparticles to obtain the so-called AP beads that showed comparable antibody enrichment as that of commercial beads. Meanwhile, the co-assembly of homo-PEN with red-emitting fluorescent quantum dots (QD) leads to generation of the Q-beads exhibiting stable fluorescence both in aqueous solution and after internalized into macrophage cells, which can be explored for quantitative evaluation of macrophage phagocytosis activity via flow cytometry. In short, this work opens new way for fabrication of multifunctional bioactive microparticles using easily synthesized polyarylene ethers.

Suppression of Extracellular Vesicle CD47 Induces Systemic Anti-DLBCL Immunity

Purpose:Tumor cells evade the immune surveillance by up-regulating surface expression of CD47, which interacts with SIRPa on macrophages to elicit the immune checkpoint response. Anti-CD47 antibody (IBI188) has shown promise in treating tumors, including Diffuse large B cell lymphoma (DLBCL). The objective response rate among DLBCL was 73% (ClinicalTrials.gov number, NCT02953509.). The basis of differential therapeutic success between patients remains unknown. Extracellular vesicles (EVs) carry bioactive molecules that influence the immune system. CD47 has be found on surface of EVs. The purpose of this study is to explore whether EV CD47 contributes to immunosuppression and is associated with anti-CD47 response in DLBCL.Methods: 1.Kaplan-Meier curves and Cox regression models were used to analyze PFS and OS. The area under the ROC curve (AUC) was used to assess the predicted validity of the National Comprehensive Cancer Network-International Prognostic Index (NCCN-IPI) model and the NCCN-IPI+ CD47 model.2.Immunofluorescence experiment was used to detect the infiltration of M1 and M2 subtype macrophages in DLBCL tumor tissue. M1 and M2 macrophages were generated from peripheral blood mononuclear cells obtained from healthy donors. Immunoelectron microscopy, flow cytometry and western blot were performed to detect CD47 on the surface of the EVs derived from DLBCL cells. The activity of macrophage phagocytosis of DLBCL cells was detected by confocal-based phagocytosis assay and flow cytometry-based phagocytosis assay.3.In DLBCL xenografts, the IBI188 antibody was a macrophage immune checkpoint inhibitor blocking CD47 that induces tumor-cell phagocytosis.4.Peripheral blood mononuclear cells from DLBCL patients and healthy donors was collected for EVs purification and subsequent detection of human CD47 proteins by ELISA.Results 1.The elevated expression of CD47 in DLBCL is significantly correlated with poor PFS and OS in a univariate analysis, and is statistically significant after adjusting for the NCCN-IPI in the univariate and multivariate analysis (Figure 1A, B). AUC analysis with cross-validation showed that the combination of the CD47 signature and NCCN-IPI had a better prediction of PFS and OS than without the CD47 signature (Figure 1C). Thus, CD47 in DLBCL can be used as a biomarker of prognosis and developed the prognostic stratification of DLBCL patients.2.EV CD47 derived from DLBCL cells has the same membrane topology as the cell surface CD47, with its extracellular domain exposed on the surface of the EVs (Figure 1D-G). Moreover, EV CD47 could function similarly to cell-surface CD47 in the suppression of macrophage phagocytosis of DLBCL cells (Figure 1H, I). DLBCL cells that had higher levels of cellular CD47 protein packaged greater amounts of CD47 into EVs, and these EVs displayed an increased binding to SIRPa of macrophages, thus inhibiting macrophage-mediated phagocytosis of DLBCL cells (Figure 1J-O).3.Antibodies against human CD47 specifically identified human CD47 on the circulating exosomes from mice bearing DLBCL xenografts but not the control mice. The level of circulating EV CD47 positively correlated with tumor size. Injection of EVs derived from DLBCL cells promoted the growth of tumors, whereas pre-treatment of the EVs with anti-CD47 antibodies (IBI188), but not IgG isotype or CD63-blocking antibodies, inhibited the effect (Figure 1P, Q).4.The presence of CD47 in the EVs isolated from the plasma of DLBCL patients, and the level of CD47 on the circulating EVs was significantly higher in DLBCL patients than that in healthy donors (Figure 1R, S). The receiver ROC curve shows that the level of circulating EV CD47 could distinguish DLBCL patients from healthy donors.ConclusionOur studies suggest that the EV CD47-SIRPa interaction may represent a critical mechanism by which DLBCL cells escape immune-mediated clearance. Our study also raises the possibility that disrupting the interaction between the EV CD47 and macrophage SIRPa is a mechanism in the CD47-SIRPa blockade-based therapies. Moreover, high levels of circulating EV CD47 would follow and correlate positively with the phagocytic activity of macrophages, and reflect the presence of a successful anti-tumor immunity elicited by the anti-CD47 therapy. Together, these findings show that EV CD47 represents an unexplored therapeutic target, which could overcome resistance to current CD47 antibody approaches. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Immunostimulatory effects of asiatic acid and madecassoside on the phagocytosis activities of macrophages cell line (J774A.1)

Macrophages play a vital role in primary immune responses mainly by carrying out phagocytosis that involves the production of nitric oxide (NO). NO is synthesized by inducible nitric oxide synthase (iNOS) to combat aetiological agents that are able to cause infectious diseases (IDs). Asiatic acid (AA) and madecassoside (MA) are two pentacyclic triterpene compounds derived from Centella asiatica (Linn.) Urban that involved in many pharmacological activities of this plant. However, there is still limited study using a normal immune model that resembles a healthy community to test the immunomodulatory effect of these compounds. Thus, this study used unstimulated macrophages as an in vitro normal model to investigate the effect of both compounds on phagocytosis activity. To achieve the aim of this study, the effects of AA and MA, alone and in combination, on the phagocytosis uptake, NO production, and iNOS expression of macrophages after 24 hours of treatment were measured. Briefly, macrophages (J774A.1) were cultured in five groups, namely untreated macrophages, macrophages treated with lipopolysaccharides (LPS) as a positive control, and macrophages treated with AA and MA alone and in combination (AA + MA). The investigation on phagocytosis uptake, NO production, and iNOS expression were conducted by using microscopic analysis on Giemsa-stained slides, the Griess assay, and Western blot, respectively. The results presented that the combination (AA + MA) treatment enhanced phagocytosis uptake, NO production, and iNOS expression of macrophages when compared to the untreated macrophages as well as macrophages treated with AA and MA, respectively. In conclusion, the combination (AA + MA) treatment is able to stimulate phagocytosis activity of macrophages and this new finding provides initial knowledge for the development of a natural product-based preventive agent against IDs.

Taurolithocholic acid but not tauroursodeoxycholic acid rescues phagocytosis activity of bone marrow-derived macrophages under inflammatory stress.

Spinal cord injury (SCI) causes cell death and consequently the breakdown of axons and myelin. The accumulation of myelin debris at the lesion site induces inflammation and blocks axonal regeneration. Hematogenous macrophages contribute to the removal of myelin debris. In this study, we asked how the inflammatory state of macrophages affects their ability to phagocytose myelin. Bone marrow‐derived macrophages (BMDM) and Raw264.7 cells were stimulated with lipopolysaccharides (LPS) or interferon gamma (IFNγ), which induce inflammatory stress, and the endocytosis of myelin was examined. We found that activation of the TLR4‐NFκB pathway reduced myelin uptake by BMDM, while IFNγ‐Jak/STAT1 signaling did not. Since bile acids regulate lipid metabolism and in some cases reduce inflammation, our second objective was to investigate whether myelin clearance could be improved with taurolithocholic acid (TLCA), tauroursodeoxycholic acid or hyodeoxycholic acid. In BMDM only TLCA rescued myelin phagocytosis, when this activity was suppressed by LPS. Inhibition of protein kinase A blocked the effect of TLCA, while an agonist of the farnesoid X receptor did not rescue phagocytosis, implicating TGR5‐PKA signaling in the effect of TLCA. To shed light on the mechanism, we measured whether TLCA affected the expression of CD36, triggering receptor on myeloid cells‐2 (TREM2), and Gas6, which are known to be involved in phagocytosis and affected by inflammatory stimuli. Concomitant with an increase in expression of tumour necrosis factor alpha, LPS reduced expression of TREM2 and Gas6 in BMDM, and TLCA significantly diminished this downregulation. These findings suggest that activation of bile acid receptors may be used to improve myelin clearance in neuropathologies.

Study on the Enhancement of Immune Function of Astaxanthin from Haematococcus pluvialis.

This study was aimed at investigating the effect of astaxanthin on the immune function and its safety in mice. It was administered once daily at low, medium and high doses (4.2, 8.35, 16.70 mg/kg BW) to mice for 30 days. Subsequently, the spleen and thymus index, spleen lymphocyte transformation activity, delayed allergy reaction, amounts of antibody-producing cells, half-hemolytic value HC50, carbon particle clearance rate, macrophage phagocytosis, and natural killer cell (NK) activity were determined. Acute oral toxicity and genotoxicity tests were conducted to evaluate the safety of astaxanthin. Compared with the control group, medium and high doses of astaxanthin significantly increased the proliferation and transformation activities of spleen lymphocytes, activities of antibody-producing cells, serum hemolysin levels, and carbon particle clearance rate in mice (phagocytic index). High doses significantly improved delayed allergy reaction and NK cell activity. Results of acute oral toxicity and genotoxicity tests were negative. Gross anatomical observations and histopathological examination showed no abnormal changes associated with the treatments. In the article, it is confirmed that astaxanthin treatments significantly improve immune functions and show no toxic effects in the experimental doses.

Novel insights into the immune regulatory effects of Megalobrama amblycephala intelectin on the phagocytosis and killing activity of macrophages

Previous studies have found that the expression level of Megalobrama amblycephala intelectin (MaINTL) increased significantly post Aeromonas hydrophila infection, and recombinant MaINTL (rMaINTL) protein could activate macrophages and enhance the phagocytosis and killing activity of macrophages. In order to reveal the immune regulatory mechanisms of MaINTL, primary M. amblycephala macrophages were treated with endotoxin-removed rMaINTL and GST-tag proteins, then total RNA were extracted and used for comparative Digital Gene Expression Profiling (DGE). 1247 differentially expressed genes were identified by comparing rMaINTL and GST-tag treated macrophage groups, including 482 up-regulated unigenes and 765 down-regulated unigenes. In addition, eleven randomly selected differentially expressed genes were verified by qRT-PCR, and most of them shared the similar expression patterns as that of DGE results. GO enrichment revealed that the differentially expressed genes were mainly concentrated in the membrane part and cytoskeleton of cellular component, the binding and signal transducer activity of molecular function, the cellular process, regulation of biological process, signaling and localization of biological process, most of which might related with the phagocytosis and killing activity of macrophages. KEGG analysis revealed the activation and involvement of differentially expressed genes in immune related pathways, such as Tumor necrosis factor (TNF) signaling pathway, Interleukin 17 (IL-17) signaling pathway, Toll-like receptor signaling pathway, and NOD like receptor signaling pathway, etc. In these pathways, TNF-ɑ, Activator protein-1 (AP-1), Myeloid differentiation primary response protein MyD88 (MyD88), NF-kappa-B inhibitor alpha (ikBɑ) and other key signaling factors were significantly up-regulated. These results will be helpful to clarify the immune regulatory mechanisms of fish intelectin on macrophages, thus providing a theoretical basis for the prevention and control of fish bacterial diseases.

Notch Signaling Ligand Jagged1 Enhances Macrophage-Mediated Response to Helicobacter pylori.

Helicobacter pylori (H. pylori) is one of the gram-negative bacteria that mainly colonize the stomach mucosa and cause many gastrointestinal diseases, such as gastritis, peptic ulcer, and gastric cancer. Macrophages play a key role in eradicating H. pylori. Recent data have shown that Notch signaling could modulate the activation and bactericidal activities of macrophages. However, the role of Notch signaling in macrophages against H. pylori remains unclear. In the present study, in the co-culture model of macrophages with H. pylori, the inhibition of Notch signaling using γ-secretase decreased the expression of inducible nitric oxide synthase (iNOS) and its product, nitric oxide (NO), and downregulated the secretion of pro-inflammatory cytokine and attenuated phagocytosis and bactericidal activities of macrophages to H. pylori. Furthermore, we identified that Jagged1, one of Notch signaling ligands, was both upregulated in mRNA and protein level in activated macrophages induced by H. pylori. Clinical specimens showed that the number of Jagged1+ macrophages in the stomach mucosa from H. pylori-infected patients was significantly higher than that in healthy control. The overexpression of Jagged1 promoted bactericidal activities of macrophages against H. pylori and siRNA-Jagged1 presented the opposite effect. Besides, the addition of exogenous rJagged1 facilitated the pro-inflammatory mediators of macrophages against H. pylori, but the treatment of anti-Jagged1 neutralizing antibody attenuated it. Taken together, these results suggest that Jagged1 is a promoting molecule for macrophages against H. pylori, which will provide insight for exploring Jagged1 as a novel therapeutic target for the control of H. pylori infection.

ISOLASI SENYAWA AKTIF DAN UJI AKTIVITAS EKSTRAK JAHE MERAH (ZINGIBER OFFICINALE) SEBAGAI IMUNOMODULATOR

Red ginger (Zingiber officinale) contains many chemical compounds that have many activities. This study aims to determine the active compounds of Zingiber officinale and immunomodulatory activity that can increase the phagocytosis of macrophage cells and lymphocyte cell proliferation. Red ginger extract compounds were identified using GC-MS. The parameters of immunostimulatory activity were macrophage phagocytosis and lymphocyte proliferation, the preparations were made in several extract concentrations. This research is an experimental study in the laboratory. The research included fresh red ginger, made into ethanol extract, dried. The results were made a suspension, the extract was then given to macrophage cells (phagocytosis testing) and lymphocyte cells (proliferation testing) in vitro to test the immunomodulatory effect. The results showed that the most active extract in macrophage phagocytosis activity was the extract concentration of 500 ppm. While the activity of lymphocyte proliferation was shown by the concentration of group A (Cell, 500 ppm extract, MK, Engerik). The active compound contained in the ethanol extract of red ginger is thought to be Geraniol

Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis

Paratuberculosis is a disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Vaccination is the most cost-effective control method. However, despite the fact that macrophages are the main target cells for this pathogen, the precise mechanisms behind the response of the macrophage to Map infection and how it is modified by vaccination are yet poorly understood. The aim of this study was to investigate the effect of Silirum® vaccination in the early immune response of caprine monocyte-derived macrophages (CaMØs). Peripheral blood mononuclear cells (PBMCs) were obtained from vaccinated and non-vaccinated goats, cultured in vitro until differentiation to macrophages and infected with Map. After a 24 h incubation, Map viability and DNA were assessed in culture by viable colony count and real time quantitative polymerase chain reaction (qPCR). In addition, Map phagocytosis and expression of IL-10, IL-12, IFN-γ, TNF-α, IL-17A, IL-1β, iNOS, IL-6 and MIP-1β were also evaluated through immunofluorescence labelling and reverse transcriptase qPCR (RT-qPCR), respectively. A significant reduction of Map viability was observed in both supernatants (P < 0.05) and CaMØs (P < 0.001) from the vaccinated group. Similarly, the percentage of infected CaMØs and the number of internalized Map by CaMØs (P < 0.0001) was higher in the vaccinated group. Finally, iNOS (P < 0.01) and IL-10 were significantly up-regulated in CaMØs from vaccinated goats, whereas only MIP-1β was up-regulated in non-vaccinated animals (P < 0.05). These results show that vaccination modifies the immune response of CaMØs, suggesting that the phagocytosis and microbiocidal activity of macrophages against Map is enhanced after vaccination.

A study of serum phosphate levels and its correlation with curb-65 score in community acquired pneumonia

Background: Community acquired pneumonia (CAP) refers to pneumonia contracted by a person with little or no contact with health care system. Phosphorus is an essential molecule in ATP, playing a central role in energy production. A normal range is 2.5 to 4.5 mg/dl. Phosphate disturbance are noted in patients with pneumonia. Hypophosphatemia, plays a role in impairing chemotaxis, phagocytosis and bactericidal activity of macrophages. Hyperphosphatemia in pneumonia result in hypocalcaemia and pulmonary calcification. Hence study of serum phosphate levels in community acquired pneumonia is of clinical significance.&#x0D; Aims and Objective: The was aimed to estimate the levels of serum phosphate in patients with community acquired pneumonia and to correlate serum phosphate levels with CURB 65 severity score.&#x0D; Materials and Methods: Seventy-five CAP patients admitted as inpatients are included in this study. Chest X ray is obtained in all suspected cases. Serum phosphate levels are determined through blood investigation on Day 1 and Day 3.&#x0D; Results: Serum phosphate levels on Day 1 and Day 3 were significantly associated with CURB65 score. Both high and low phosphate levels were associated with high CURB 65 score. The association between phosphate levels and CURB 65 score was significant in patients who got discharged but not in patients who expired.&#x0D; Conclusions: Present study of serum phosphate levels as biomarkers in CAP showed that both hypophosphatemia and hyperphosphatemia carried poor prognosis which correlated with high CURB65 score.

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

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

Human Amnion-Derived Mesenchymal Stromal Cells in Cirrhotic Patients with Refractory Ascites: A Possible Anti-Inflammatory Therapy for Preventing Spontaneous Bacterial Peritonitis

Cirrhosis is associated with dysregulated immune cell activation and immune dysfunction. These conditions modify gut flora, facilitate bacterial translocation, and increase susceptibility to bacterial peritonitis and consequent systemic infections by dramatically affecting long-term patient survival. Human amnion-derived mesenchymal stromal cells (hA-MSCs) exert immunomodulatory potential benefit, and have the ability to modulate their actions, especially in situations requiring immune activation through mechanisms not fully understood. In this study, we aimed to investigate, in vitro, the immunostimulant or immunosuppressive effects of hA-MSCs on cellular components of ascitic fluid obtained from cirrhotic patients with refractory ascites. We found that hA-MSCs viability is not affected by ascitic fluid and, interestingly, hA-MSCs diminished the pro-inflammatory cytokine production, and promoted anti-inflammatory M2 macrophage polarization. Moreover, we found that there was no simultaneous significant decrease in the M1-like component, allowing a continual phagocytosis activity of macrophages and NK cells to restore a physiological condition. These data highlight the plasticity of hA-MSCs’ immunomodulatory capacity, and pave the way to further understanding their role in conditions such as spontaneous bacterial peritonitis.Graphical abstract

In vitro Immunomodulatory Effect from Edible Green Seaweed of Caulerpa lentillifera Extracts on Nitric Oxide Production and Phagocytosis Activity of RAW 264.7 Murine Macrophage Cells

<strong>Background: </strong><em>Caulerpa lentillifera</em> is edible macroalgae which has several biological properties. This marine plant also contains beneficial secondary metabolites that can be used for treatment and prevent several deficiencies. <strong>Objectives: </strong>This research was conducted to evaluate immunomodulatory effect on nitric oxide production and phagocytosis of RAW 264.7 murine macrophage cells from edible green seaweed of <em>C. lentillifera</em> extract. <strong>Methods: </strong>Extraction was applied to obtained ethanol extract, water extract and insoluble fraction. Alamar blue inclusion method was used to determine cytotoxicity and proliferation of RAW 264.7 after treated with sample. Griess assay using colourimetric commercial kits was selected to evaluate NO production and phagocytosis effect of samples was determined using phagocytic kit. <strong>Results:</strong><em> C. lentillifera </em>extracts did not show any cytotoxic effect against RAW 264.7 macrophage. One hundred &mu;g/mL of water extract increased the production of nitrite oxide. This extract revealed suspected-giant cells formation as a results of phagocytosis activity. The extract of <em>C. lentillifera </em>also enhanced phagocytosis activity of murine macrophage against <em>E. coli. C. lentillifera</em> extract enhanced activity of RAW 264.7 murine macrophage by increasing NO production and phagocytosis activity. This extract also caused suspected-giants&rsquo; cells as indicator of phagocytosis effect. <strong>Conclusion: </strong>This result provides an insight that the potency of<em> C. lentillifera </em>extract as immunostimulant. <p style="text-align: justify;"><strong>Key words: </strong>Caulerpa lentillifera, Immunomodulatory, Macrophage, Nitric oxide, Phagocytosis.

The immunomodulatory effects of Carapax Trionycis ultrafine powder on cyclophosphamide-induced immunosuppression in Balb/c mice.

There are abundant resources of Carapax Trionycis from soft-shelled turtle processing wastes each year in China. Our preliminary work showed that Carapax Trionycis ultrafine powder (CTUP) obtained using ball-milling with a particle size of 2.24 μm (D0.025) contained more active ingredients. The CTUP D0.025 has a good bioaccessibility, but there has been no report about the immunomodulatory function of CTUP. Therefore, using a cyclophosphamide-induced immunosuppression mice model, we investigated the immunomodulatory effects of CTUP D0.025. The results indicated that CTUP D0.025 administration significantly improved the immune organ (bone marrow, thymus and spleen) indices, ameliorated spleen tissue morphology and increased the capacity of splenocyte proliferation and the activity of macrophage phagocytosis. CTUP D0.025 also significantly promoted the secretion of cytokines (IL-2, IL-4, IL-10, IFN-γ and TNF-α), improved the related mRNA expression levels of IL2, IFN-γ, T-bet and GATA3 in immunosuppressed mice and increased the production of serum hemolysin and the levels of IgG, IgM as well as complement C3 . Moreover, CTUP D0.025 administration enhanced the antioxidant capacity of mice, exhibited a moderating effect on the damage of bone and skeletal muscle and improved the recovery of bone mineral density and calcium metabolism. These findings demonstrated that CTUP D0.025 had an effective immune-enhancing function in immunosuppressive Balb/c mice and also exhibited anti-osteoporosis properties. © 2020 Society of Chemical Industry.