Immunostimulatory Activity Research Articles

Structural characterization of polysaccharides from Dictyophora rubrovolvata mycelium and their immunostimulatory activity in RAW264.7 cells

In this study, a homogeneous polysaccharide, named, DRMP-1 was isolated from the mycelium of Dictyophora rubrovolvata by water extraction and alcohol precipitation, and its structure and activity were studied. The molecular weight of DRMP-1 is 2.33 × 106 Da, and it does not contain protein and nucleic acid. DRMP-1 is composed of mannose, glucose and galactose with a molar ratio of 0.04: 1: 0.04. Through methylation and NMR analysis, DRMP-1 is mainly composed of 1 → 6)-α-Galp-( 1 →, T-α-D-Glcp, →1)-α-D-Manp, → 6)-β-D-Glcp-( 1 →, → 4, 6)- α-D-Manp-( 1 →, → 4)- α-D-Glcp-( 1 →, → 3)-β-D-Glcp-( 1 → and → 4, 6)- α-D-Glcp-( 1 →. In addition, the immunomodulative activity of DRMP-1 was evaluated by establishing the RAW264.7 cell model. The results showed that DRMP-1 could significantly increase phagocytosis activity and NO production more than twice as much as that of the blank group, and could increase intracellular ACP, LSZ and SOD activities. Therefore, it is concluded that the polysaccharide of Dictyophora rubrovolvata mycelium may be a potential immunomodulator.

The Eleutherococcus senticosus fruits' intractum affects changes in the transepithelial electric potential in the distal section of the rabbit's large intestine and inhibits hyaluronidase

Ethnopharmacological relevanceEleutherococcus senticosus (Rupr. et Maxim.) Maxim. has been used in traditional Russian medicine due to its recognized immunostimulant and anti-inflammatory activities. Compounds present in the fruits have demonstrated the capability to modulate the activity of enzymes such as hyaluronidase, suggesting their potential value in the development of effective therapies for various conditions where anti-inflammatory properties are beneficial, such as gastrointestinal diseases and tumor growth. Aim of the studyIn order to support the use of the fruits in folk medicine, this study is aimed to evaluate, post-mortem, the impact of E. senticosus fruits intractum (40 % extract made from fresh fruits) on the transepithelial electrogenic transport of sodium ions in the colon. The objective of this study was also to examine the impact of the intractum on proinflammatory serum hyaluronidase in children diagnosed with acute leukemia. MethodsThe study employed the Ussing technique to examine electrophysiological characteristics of isolated epithelial tissue, using the distal colon wall isolated from 10 New Zealand white male rabbits. The effect of the intractum on the inhibition of human serum hyaluronidase was examined with turbidimetric screening methods, using the blood samples collected from patients diagnosed with acute leukemia. ResultsFor the first time, we discovered that the intractum used in the stimulation fluid, caused hyperpolarization reactions in colon tissue. Statistical analysis showed that these reactions were significantly different in relation to the control. The intractum significantly inhibited hyaluronidase activity with the mean value by group of 60 %, and 40 % for aescin used as a control. ConclusionThe results support the traditional use of the fruits in inflammatory-related diseases. The use of intractum of E. senticosus on the distal colon wall demonstrates its protective effect on the wall integrity and in a relation to hyaluronidase inhibition may additionally indicate its anti-inflammatory property. Thus, the results mean that the intractum may be used in colon-related diseases.

Immunostimulant and Pharmacological Activities, of Cucurbita Maxima Seeds on Humoral and Cellular Immunity Cells and their Functions

Background Natural resources are key for discovering medicinal compounds. Hence, the objective of this study was to evaluate the immunomodulatory activities of Cucurbita maxima (pumpkin) seeds, a component of Moroccan folk medicine believed to enhance immunity. Methods We produced three distinct extracts employing ethanol, ethyl acetate, and water as solvents. The immunomodulatory effects of these extracts were assessed on rabbit immune cell proliferation and their functions, including IgG production, cytotoxicity, and phagocytosis. Results The study findings reveal a significant stimulation in thymocyte proliferation, yielding a remarkable 285.7% response, along with a 146.5% increase in cytotoxicity (MLR) in response to the aqueous extract. Furthermore, there was a notable enhancement in complement activity by 119.9% in response to the aqueous extract, implying the potential of C. maxima to modulate both humoral and cellular immunity. Additionally, both EtOAc and aqueous extracts led to a reduction in macrophage phagocytic function. The assessment of the antibacterial properties of ethanol extract showed greater effectiveness against Bacillus, E. Coli, and Staphylococcus. In a separate analysis, antioxidant activity was gauged through DPPH, FRAP, and ABTS methods. These tests exhibited robust antioxidative effects across all extracts, characterized by higher levels of phenolic and flavonoid content. The FT-IR spectrum indicated the presence of compounds such as triglycerides, sterol esters, phospholipids, and unsaturated fatty acids in the extracts. Conclusion Collectively, these outcomes prompt the consideration of utilizing the aqueous extract of Cucurbita maxima to strengthen humoral and cellular immunity, as well as lymphocyte toxicity, in various ailments.

Complete biosynthesis of the potent vaccine adjuvant QS-21

QS-21 is a potent vaccine adjuvant currently sourced by extraction from the Chilean soapbark tree. It is a key component of human vaccines for shingles, malaria, coronavirus disease 2019 and others under development. The structure of QS-21 consists of a glycosylated triterpene scaffold coupled to a complex glycosylated 18-carbon acyl chain that is critical for immunostimulant activity. We previously identified the early pathway steps needed to make the triterpene glycoside scaffold; however, the biosynthetic route to the acyl chain, which is needed for stimulation of T cell proliferation, was unknown. Here, we report the biogenic origin of the acyl chain, characterize the series of enzymes required for its synthesis and addition and reconstitute the entire 20-step pathway in tobacco, thereby demonstrating the production of QS-21 in a heterologous expression system. This advance opens up unprecedented opportunities for bioengineering of vaccine adjuvants, investigating structure–activity relationships and understanding the mechanisms by which these compounds promote the human immune response.

Polymeric nanocapsules loaded with poly(I:C) and resiquimod to reprogram tumor-associated macrophages for the treatment of solid tumors.

In the tumor microenvironment (TME), tumor-associated macrophages (TAMs) play a key immunosuppressive role that limits the ability of the immune system to fight cancer. Toll-like receptors (TLRs) ligands, such as poly(I:C) or resiquimod (R848) are able to reprogram TAMs towards M1-like antitumor effector cells. The objective of our work has been to develop and evaluate polymeric nanocapsules (NCs) loaded with poly(I:C)+R848, to improve drug stability and systemic toxicity, and evaluate their targeting and therapeutic activity towards TAMs in the TME of solid tumors. NCs were developed by the solvent displacement and layer-by-layer methodologies and characterized by dynamic light scattering and nanoparticle tracking analysis. Hyaluronic acid (HA) was chemically functionalized with mannose for the coating of the NCs to target TAMs. NCs loaded with TLR ligands were evaluated in vitro for toxicity and immunostimulatory activity by Alamar Blue, ELISA and flow cytometry, using primary human monocyte-derived macrophages. For in vivo experiments, the CMT167 lung cancer model and the MN/MCA1 fibrosarcoma model metastasizing to lungs were used; tumor-infiltrating leukocytes were evaluated by flow cytometry and multispectral immunophenotyping. We have developed polymeric NCs loaded with poly(I:C)+R848. Among a series of 5 lead prototypes, protamine-NCs were selected based on their physicochemical properties (size, charge, stability) and in vitro characterization, showing good biocompatibility on primary macrophages and ability to stimulate their production of T-cell attracting chemokines (CXCL10, CCL5) and to induce M1-like macrophages cytotoxicity towards tumor cells. In mouse tumor models, the intratumoral injection of poly(I:C)+R848-protamine-NCs significantly prevented tumor growth and lung metastasis. In an orthotopic murine lung cancer model, the intravenous administration of poly(I:C)+R848-prot-NCs, coated with an additional layer of HA-mannose to improve TAM-targeting, resulted in good antitumoral efficacy with no apparent systemic toxicity. While no significant alterations were observed in T cell numbers (CD8, CD4 or Treg), TAM-reprogramming in treated mice was confirmed by the relative decrease of interstitial versus alveolar macrophages, having higher CD86 expression but lower CD206 and Arg1 expression in the same cells, in treated mice. Mannose-HA-protamine-NCs loaded with poly(I:C)+R848 successfully reprogram TAMs in vivo, and reduce tumor progression and metastasis spread in mouse tumors.

Exploration of the profiles of steroidal saponins from Rhizoma Paridis and their metabolites in rats by UPLC-Q-TOF-MS/MS.

Steroidal saponins characterised by intricate chemical structures are the main active components of a well-known traditional Chinese medicine (TCM) Rhizoma Paridis. The metabolic profiles of steroidal saponins in vivo remain largely unexplored, despite their renowned antitumor, immunostimulating, and haemostatic activity. To perform a comprehensive analysis of the chemical constituents of Rhizoma Paridis total saponins (RPTS) and their metabolites in rats after oral administration. The chemical constituents of RPTS and their metabolites were analysed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS). A reliable UPLC-Q-TOF-MS/MS method was established, and a total of 142 compounds were identified in RPTS. Specifically, diosgenin-type saponins showed the diagnostic ions at m/z 415.32, 397.31, 283.25, 271.21, and 253.20, whereas pennogenin-type saponins exhibited the diagnostic ions at m/z 413.31, 395.30, and 251.20. Based on the characteristic fragments and standard substances, 15 specific metabolites were further identified in the faeces, urine, plasma, and bile of rats. The metabolic pathways of RPTS, including phase I reactions (de-glycosylation and oxidation) and phase II reactions (glucuronidation), were explored and summarised, and the enrichment of metabolites was characterised by multivariate statistical analysis. The intricate RPTS could be transformed into relatively simple metabolites in rats through de-glycosylation, which provides a reference for further metabolic studies and screening of active ingredients for TCM.

Combination Therapy with a TLR7 Agonist and a BRD4 Inhibitor Suppresses Tumor Growth via Enhanced Immunomodulation.

JQ-1 is a typical BRD4 inhibitor with the ability to directly fight tumor cells and evoke antitumor immunity via reducing the expression of PD-L1. However, problems arise with the development of JQ-1 in clinical trials, such as marked lymphoid and hematopoietic toxicity, leading to the investigation of combination therapy. SZU-101 is a TLR7 agonist designed and synthesized by our group with potent immunostimulatory activity. Therefore, we hypothesized that combination therapy of SZU-101 and JQ-1 would target innate immunity and adaptive immunity simultaneously, to achieve a better antitumor efficacy than monotherapy. In this study, the repressive effects of the combination administration on tumor growth and metastasis were demonstrated in both murine breast cancer and melanoma models. In 4T1 tumor-bearing mice, i.t. treatment with SZU-101 in combination with i.p. treatment with JQ-1 suppressed the growth of tumors at both injected and uninjected sites. Combination therapy increased M1/M2 ratio in TAMs, decreased PD-L1 expression and promoted the recruitment of activated CD8+ T cells in the TME. In summary, the improved therapeutic efficacy of the novel combination therapy appears to be feasible for the treatment of a diversity of cancers.

Structure and immunostimulatory activity studies on two novel Flammulina velutipes polysaccharides: revealing potential impacts of →6)-α-D-Glcp(1→ on the TLR-4/MyD88/NF-κB pathway.

Two novel Flammulina velutipes (F. velutipes) polysaccharides, FVPH1 and FVPH2, were isolated and purified after hot water extraction. The structural characterization revealed that the backbone of FVPH1 consisted mainly of →6)-α-D-Glcp(1→, →3,4)-α-D-Galp(1→, →4)-α-L-Fucp(1→, and →4)-β-D-Manp(1→, while the backbone of FVPH2 consisted of →3)-α-D-Galp(1→, →3,4)-α-D-Manp(1→,→6)-α-D-Glcp(1→. The branches of FVPH1 contained →6)-α-D-Glcp(1→ and α-D-Glcp(1→ and the branches of FVPH2 consisted of →3)-α-D-Galp(1→, →6)-α-D-Glcp(1→, and β-L-Fucp(1→. FVPH2 exhibited significantly better immunostimulatory activity than FVPH1 (P < 0.05), as evidenced by the increased expression of NO, IL-1β, IL-6, and TNF-α and pinocytic activity of RAW264.7 cells. As the most abundant structure in the polysaccharides of F. velutipes, the content of →6)-α-D-Glcp(1→ might play a crucial role in influencing the immunostimulatory activity of F. velutipes polysaccharides. The F. velutipes polysaccharide with a lower content of →6)-α-D-Glcp(1→ and a higher branching degree could significantly enhance the immunostimulatory activity of F. velutipes polysaccharides via activating the TLR-4/MyD88/NF-κB pathway more effectively.

High productivity of immunostimulatory membrane vesicles of Limosilactobacillus antri using glycine.

Nanosized membrane vesicles (MVs) released by bacteria play important roles in both bacteria-bacteria and bacteria-host interactions. Some gram-positive lactic acid bacteria produce MVs exhibiting immunoregulatory activity in the host. We found that both bacterial cells and MVs of Limosilactobacillus antri JCM 15950, isolated from the human stomach mucosa, enhance immunoglobulin A production by murine Peyer's patch cells. However, the thick cell walls of gram-positive bacteria resulted in low MV production, limiting experiments and applications using MVs. In this study, we evaluated the effects of glycine, which inhibits cell wall synthesis, on the immunostimulatory MV productivity of L. antri. Glycine inhibited bacterial growth while increasing MV production, with 20 g/L glycine increasing MV production approximately 12-fold. Glycine was most effective at increasing MV production when added in the early exponential phase, which indicated that cell division in the presence of glycine increased MV production. Finally, glycine increased MV productivity approximately 16-fold. Furthermore, glycine-induced MVs promoted interleukin-6 production by macrophage-like J774.1 cells, and the immunostimulatory activity was comparable to that of spontaneously produced MVs. Our results indicate that glycine is an effective agent for improving the production of MVs with immunostimulatory activity in gram-positive lactic acid bacteria, which can be applied as mucosal adjuvants and functional foods.

Stability of shark (Centrophorus sp.) liver oil rich in squalene as an immunostimulant

Shark liver oil has been widely produced in Indonesia; however, many shark liver oil products still do not meet the established standards. This research involves the addition of antioxidants to stabilize the oil as an effort to extend its shelf life. The objectives of this study were to determine the immunostimulant activity, assessed the synergistic effect of vitamin E in shark liver oil rich in squalene, and identified the optimum concentration of the antioxidant vitamin E in maintaining its stability. Immunostimulant activity was tested by observing spleenocyte proliferation, phagocytic index, and capacity in the peritoneal cavity cells of mice. The stability of the oil was examined by an accelerated method using an oven, what was called the schaal oven test at 40 °C. The research results indicate that shark liver oil possesses immunostimulant effects; however, with an increasing dosage of fish oil, there is no significant improvement. The presence of vitamin E in cod liver oil exhibited a synergistic effect on spleenocyte proliferation for all dosages, while the phagocytic index and capacity at a dosage of 100 μg/mL showed synergy. However, at a dosage of 50 μg/mL, the phagocytic index and capacity did not exhibit synergy with the immunostimulant. The optimal concentration of the antioxidant vitamin E in maintaining oil stability was found to be 100 ppm.

Algae in Biomedicine.

Algae, which live in marine or freshwater, are photosynthetic organisms. They vary greatly in size, morphology, and degree of complexity of their body structures. Algae are generally divided into two main groups, microalgae, which are small in size, and macroalgae, which are larger in size. These aquatic organisms have rich and valuable compounds including sterols, polysaccharides, pigments, fatty acids, proteins, enzymes, minerals, and vitamins that could be used in different application fields due to their bioactive functions. In recent years, algae and their components have attracted interest in biomedicine and health applications as their bioactive components could show antioxidant, anticancer, anti-inflammatory, antiviral, antiangiogenic, antidiabetic, antiobesity, immunostimulatory, vaccine adjuvant, and hypolipidemic activities. In this chapter, these activities and bioactive components underlying these properties are reviewed.

Emerging nanoparticle platforms for CpG oligonucleotide delivery.

Unmethylated cytosine-phosphate-guanine (CpG) oligodeoxynucleotides (ODNs), which were therapeutic DNA with high immunostimulatory activity, have been applied in widespread applications from basic research to clinics as therapeutic agents for cancer immunotherapy, viral infection, allergic diseases and asthma since their discovery in 1995. The major factors to consider for clinical translation using CpG motifs are the protection of CpG ODNs from DNase degradation and the delivery of CpG ODNs to the Toll-like receptor-9 expressed human B-cells and plasmacytoid dendritic cells. Therefore, great efforts have been devoted to the advances of efficient delivery systems for CpG ODNs. In this review, we outline new horizons and recent developments in this field, providing a comprehensive summary of the nanoparticle-based CpG delivery systems developed to improve the efficacy of CpG-mediated immune responses, including DNA nanostructures, inorganic nanoparticles, polymer nanoparticles, metal-organic-frameworks, lipid-based nanosystems, proteins and peptides, as well as exosomes and cell membrane nanoparticles. Moreover, future challenges in the establishment of CpG delivery systems for immunotherapeutic applications are discussed. We expect that the continuously growing interest in the development of CpG-based immunotherapy will certainly fuel the excitement and stimulation in medicine research.

Glucosylation endows nanoparticles with TLR4 agonist capability to trigger macrophage polarization and augment antitumor immunity

Carbohydrates have emerged as promising candidates for immunomodulation, however, how to present them to immune cells and achieve potent immunostimulatory efficacy remains challenging. Here, we proposed and established an effective way of designing unique glyconanoparticles that can amplify macrophage-mediated immune responses through structural mimicry and multiple stimulation. We demonstrate that surface modification with glucose can greatly augment the immunostimulatory efficacy of nanoparticles, comparing to mannose and galactose. In vitro studies show that glucosylation improved the pro-inflammatory efficacy of iron oxide nanoparticles (IONPs) by up to 300-fold, with the immunostimulatory activity of glucosylated IONPs even surpassing that of LPS under certain conditions. In vivo investigation show that glucosylated IONPs elicited increased antitumor immunity and achieved favorable therapeutic outcomes in multiple murine tumor models. Mechanistically, we proposed that glucosylation potentiated the immunostimulatory effect of IONPs by amplifying toll-like receptors 4 (TLR4) activation. Specifically, glucosylated IONPs directly interacted with the TLR4-MD2 complex, resulting in M1 macrophage polarization and enhanced antitumor immunity via activation of NF-κB, MAPK, and STAT1 signaling pathways. Our work provides a simple modification strategy to endow nanoparticles with potent TLR4 agonist effects, which may shed new light on the development of artificial immune modulators for cancer immunotherapy.

Identification of intracellular activation mechanism of rhamnogalacturonan-I type polysaccharide purified from Panax ginseng leaves in macrophages and roles of component sugar chains on activity.

This study aimed to investigate the mechanisms underlying intracellular signaling pathways in macrophages in relation to the structural features of rhamnogalacturonan (RG) I-type polysaccharide (PGEP-I) purified from Panax ginseng leaves. For this investigation, we used several specific inhibitors and antibodies against mitogen-activated protein kinase (MAPK), nuclear factor-kappa B (NF-κB), and pattern recognition receptors (PRRs). Furthermore, we investigated the roles of component sugar chains on immunostimulating activity through a sequential enzymatic and chemical degradation steps. We found that PGEP-I effectively induced the phosphorylation of several MAPK- and NF-κB-related proteins, such as p38, cJun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p65. Particularly, immunocytochemistry analysis confirmed the PGEP-I-induced translocation of p65 into the nucleus. Furthermore, the breakdown of PGEP-I side chains and main chain during sequential enzymatic and chemical degradation reduced the PGEP-I-induced macrophage cytokine secretion activity. IL-6, TNF-α, and NO secreted by macrophages are associated with several signaling pathway proteins such as ERK, JNK, and NF-κB and several PRRs such as dectin-1, CD11b, CD14, TLR2, TLR4, and SR. Thus, these findings suggest that PGEP-I exerts potent macrophage-activating effects, which can be attributed to its typical RG-I structure comprising arabinan, type II arabinogalactan, and rhamnose-galacturonic acid repeating units in the main chain.

Sulfated polysaccharide from brown algae Dictyota caribaea stimulates macrophages to an antitumoral phenotype

Fucoidans are sulfated polysaccharides capable of exerting biological activities such as antitumor and immunomodulatory effects. Previous studies demonstrated the antitumor activity of non-cytotoxic fucoidans from the seaweed Dictyota caribaea (Dc-SP) in vivo. Macrophages (Mfs) are innate immune cells capable of promoting or inhibiting tumor growth depending on the stimulus. This study aimed to evaluate the immunostimulant activity of Dc-SP on a murine Mfs cell line (RAW 264.7) in vitro. Dc-SP was assessed for its ability to modify cell viability and stimulate the production of antitumor markers on RAW 264.7 cells. Dc-SP induced an increase (p < 0.05) in the production of NO and cytokines TNF-α, IL-1β, and IL-10. The exposure of RAW 264.7 to Dc-SP also increased (p < 0.05) the expression of M1 phenotype markers such as iNOS, CD86, and MHC II. Dc-SP did not exhibit direct cytotoxicity on a murine melanoma cell line (B16–F10). However, the conditioned medium (CM) obtained from RAW 264.7 previously stimulated with Dc-SP (CM-Dc-SP) showed antiproliferative activity on tumor cells. B16–F10 incubated with CM-Dc-SP showed a cytostatic profile, tumor cells did not alter membrane integrity, however, they suffered morphological changes such as cell shrinkage and high granularity. In conclusion, Dc-SP stimulated RAW 264.7 to an antitumor phenotype.

Immunomodulatory Activity Analyses of Cell-Free Supernatant of Lactobacillus plantarum LP299v Strain in RAW 264.7 Macrophage Cells

The human immune system is essential for defending the body against harmful internal and external elements; immunity includes innate and acquired immunity. Macrophages, the innate immune system's key components, are crucial for the clearance of dead cells and tumor cells, as well as foreign substances by triggering phagocytosis. They also play a role in the adaptive response with the cytokines and mediator molecules they secrete. Lactic acid bacteria (LAB), an important probiotics class, have a strong potential to improve host health and can be used as a functional food. There have been reports of certain LAB strains having immunostimulating effects. However, the effects of cell-free supernatants (CFS) gathered from some LAB strains on macrophage activation have become an important research area in recent years. This study's main objective was to characterize the immunostimulatory activities of Lactobacillus plantarum LP299v in the RAW 264.7 macrophage cell line. For this purpose, the immunomodulatory activity of CFS of the related strain was evaluated by MTT, neutral red assay, and Griess reaction respectively, in terms of proliferation, phagocytosis ability, and nitric oxide (NO) production parameters using the macrophage cell line. Studies have shown that this strain significantly increased proliferation, phagocytosis, and NO levels in RAW264.7 macrophage cells. When considered, these results suggest that the cell-free supernatant, obtained from Lactobacillus plantarum LP299v selected in this study, may be helpful for candidate compounds with immunostimulatory activity.

Enhancement of Immune Functions by Limosilactobacillus reuteri KBL346: In Vitro and In Vivo Studies.

Lactobacilli have been widely used as probiotics because of their benefits for intestinal health and physiological functions. Among a variety of Lactobacillus genera, Limosilactobacillus reuteri has been studied for its ability to exert anti-inflammatory functions and its role in controlling metabolic disorders, as well as the production of the antimicrobial compound reuterin. However, the effects and mechanisms of L. reuteri on enhancing immune responses in the immunosuppressed states have been relatively understudied. In this study, we isolated an immunomodulatory strain, namely, L. reuteri KBL346 (KBL346), from a fecal sample of a 3-month-old infant in Korea. We evaluated the immunostimulatory activity and hematopoietic function of KBL346 in macrophages and cyclophosphamide (CPA)-induced immunosuppressed mice. KBL346 increased the phagocytic activity against Candida albicans MYA-4788 in macrophages, and as biomarkers for this, increased secretions of nitric oxide (NO) and prostaglandin E2 (PGE2) were confirmed. Also, the secretions of innate cytokines (TNF-α, IL-1β, and IL-6) were increased. In CPA-induced immunosuppressed mice, KBL346 at a dosage of 1010 CFU/kg protected against spleen injury and suppressed levels of immune-associated parameters, including NK cell activity, T and B lymphocyte proliferation, CD4+ and CD8+ T cell abundance, cytokines, and immunoglobulins in vivo. The effects were comparable or superior to those in the Korean red ginseng positive control group. Furthermore, the safety assessment of KBL346 as a probiotic was conducted by evaluating its antibiotic resistance, hemolytic activity, cytotoxicity, and metabolic characteristics. This study demonstrated the efficacy and safety of KBL346, which could potentially be used as a supplement to enhance the immune system.

IMMUNOSTIMULANT ACTIVITY OF HANTAP (STERCULIA COCCINEA JACK) LEAVES EXTRACT ON NON-SPECIFIC AND SPESIFIC IMMUNE RESPONSES

Objective: This study aims to determine the immunostimulant effect and the effective dose of hantap leaf ethanolic extract against non-specific and specific immune responses.&#x0D; Methods: The immunostimulant activity of the extract was tested by the carbon clearance method (non-specific response). The effect of the extract on delay-type hypersensitivity/DTH (cellular-specific response) was determined by the paw edema method. The number of leukocyte cells and the percentage of leukocyte cell types were also calculated. The antibody titer (humoral-specific response) test was carried out by the hemagglutination method.&#x0D; Results: The results of the non-specific immune response demonstrated that three doses of hantap leaf extract, when compared to the negative control, could accelerate the rate of carbon elimination and increase the phagocytosis index. Hantap leaf extract can provide an immunomodulatory effect by increasing the delayed-type hypersensitivity response by showing a greater volume of leg swelling than the negative control received CMC Na 0.5%, which was significantly different (p&lt;0.05), increasing leukocyte count and leukocyte differential and increasing primary and secondary antibody titers. The effective dose of hantap leaves extract was 200 mg/kg body weight.&#x0D; Conclusion: This study proves that hantap leaf extract has an immunomodulatory effect that increases the immune system (immunostimulant)

Atypical activation of the RNA sensor MDA5 by hepatitis C virus.

Hepatitis C virus (HCV) is a significant human pathogen that can cause a number of serious diseases including chronic inflammation of the liver, cirrhosis, and hepatocellular carcinoma. A key enzyme in the HCV life cycle is the nonstructural protein 5B (NS5B), which functions as an RNA-dependent RNA polymerase (RdRp) responsible for replicating the viral RNA genome. In their recent study, Dansako and colleagues showed that HCV NS5B induces type I interferon via activation of the RNA receptor MDA5, an activity that was dependent on the RdRp enzymatic activity but independent of viral RNA replication. Their data further indicated that the NS5B enzymes of HCV and the related GB virus-B produce cellular double-stranded RNA (dsRNA) species with potential immunostimulatory activity. These findings unveil an unconventional mechanism of activation of MDA5-mediated host immunity by viral RdRp enzymes, which is expected to spur new research directions in viral immunology.

Activation of Immune and Antiviral Effects by Euglena Extracts: A Review.

Influenza is an acute respiratory illness caused by influenza virus infection, which is managed using vaccines and antiviral drugs. Recently, the antiviral effects of plants and foods have gained attention. Euglena is a motile unicellular alga and eukaryotic photosynthetic microorganism. It has secondary chloroplasts and is a mixotroph able to feed by photosynthesis or phagocytosis. This review summarizes the influenza treatment effects of Euglena from the perspective of a functional food that is attracting attention. While it has been reported that Euglena contributes to suppressing blood sugar levels and ameliorates symptoms caused by stress by acting on the autonomic nervous system, the immunostimulatory and antiviral activities of Euglena have also been reported. In this review, I focused on the immunostimulation of antiviral activity via the intestinal environment and the suppression of viral replication in infected cells. The functions of specific components of Euglena, which also serves as the source of a wide range of nutrients such as vitamins, minerals, amino acids, unsaturated fatty acids, and β-1,3-glucan (paramylon), are also reviewed. Euglena has animal and plant properties and natural compounds with a wide range of functions, providing crucial information for improved antiviral strategies.