Immunomodulatory Activity Research Articles

Structural Characterization, and Antioxidant, Hypoglycemic and Immunomodulatory Activity of Exopolysaccharide from Sanghuangporus sanghuang JM-1

Sanghuang as a medicinal fungus in China has a history of more than 2000 years, and is known as the “forest gold”. Most notably, the polysaccharides of Sanghuangporus sp. have attracted widespread attention due to their significant bioactivity in recent years. At present, extensive studies are being carried out on the extraction methods, structural characterization, and activity evaluation of polysaccharides. Here, we aimed to evaluate the structure and bioactivity of LEPS-1, an exopolysaccharide derived from the S. sanghuang JM-1 strain. The structure was elucidated by chromatography/spectral methods and hydrolyzation, and the solubility, the antioxidant activity, hypoglycemic activity and immunomodulatory activity were investigated. Results showed that LEPS-1 contained a →2)-α-Manp(1→6)-α-Galp(1→[2)-α-Manp(1→]n→2,6)-α-Manp(1→6,2)-α-Manp(1→3)-α-Manp(1→ backbone substituted at the O-6 and O-2 positions with side chains. These two branching fragments were β-Manp(1→. The molecular weight of LEPS-1 is 36.131 kDa. The results of biological activity analysis suggested that LEPS-1 was easily soluble in water, with reducing capability and DPPH radical scavenging capability. Furthermore, the IC50 values of LEPS-1 against α-amylase and α-glucosidase were 0.96 mg/mL and 1.92 mg/mL. LEPS-1 stimulated RAW264.7 cells to release NO, TNF-α and IL-6 with no cytotoxicity, showing potent potential for immunomodulatory activity. These findings describe a potential natural exopolysaccharide with medicinal value and a basis for the development of S. sanghuang exopolysaccharides.

High dose cotrimoxazole treatment in patients with severe COVID-19: A randomised controlled trial

Background: Cotrimoxazole for severe COVID-19 may have a better prognosis because of its antibacterial, immunomodulatory, and anti-inflammatory activities. We examined the efficacy of high-dose cotrimoxazole therapy in terms of duration of hospital stays and reduction in mortality in severe COVID-19 infections. Methods: From May to November 2021, we conducted a double blind randomised controlled trial with two parallel groups in the COVID units of Bangabandhu Sheikh Mujib Medical University and Anwar Khan Modern Medical College Hospital, Dhaka. One group received standard therapy in addition to an oral dose of 960 mg of cotrimoxazole twice daily for 7 days (intervention group), while the other group received standard treatment and a placebo (standard group). Pre-protocol analysis was done. Results: A total of 188 patients were enrolled, but 166 completed the study. Of them, 93 were in the intervention group and 73 were in standard group. The mean ages of the groups were similar (intervention, 56.2 and standard, 59.2 years) (P=0.10). The mortality at 28 days between groups was also similar (11.8% in the intervention group and 15.0% in the standard group) (P=0.56). The hospital stay was 13.7 days for the intervention group and 13.5 days for the standard group (P=0.86). However, the reduction in C-reactive protein was statistically significant in the intervention group, with a mean decline of 23.6 mg/L (95% confidence interval, 0.5–46.7 mg/L). Conclusion: High-dose cotrimoxazole did not benefit in shortening in-hospital stay or reducing mortality at day 28 in patients with severe COVID-19. However, the decline in the C-reactive protein level was significant, necessitating further research.

Structural characterization and immunoregulatory mechanism of a low-molecular-weight polysaccharide from lotus root

The extraction of polysaccharide from lotus root was highly homogenized, and the structure of the polysaccharide was not clear. Herein, we report a hot water method combined with α-amylase that was applied to extract lotus root polysaccharide. After purified, a lotus root polysaccharide fraction LP60-a with high purity and low molecule weight was obtained. Systematic characterization of the structure of LP60-a was achieved by monosaccharide composition, methylation and NMR analysis, showing that LP60-a was composed of α-1,6-glucan linked with a small amount of arabinogalactan. Conformational determination showed that LP60-a was a three-helix polysaccharide with random coil conformation. Furtherly, the immunomodulatory activities of LP60-a were investigated in RAW264.7 macrophages. The data indicated that LP60-a could enhance the proliferation and phagocytosis of macrophages significantly, and induce the expression of NO and TNF-α in macrophages without causing inflammation. Moreover, LP60-a promoted the phosphorylation of MAPK p38 and JNK, as well as NF-κB p65, indicating that LP60-a could activate RAW264.7 cells through MAPK and NF-κB signaling pathways. In conclusion, the results imply that LP60-a could enhance the immune function of macrophages, presenting a possibility to play a role as an immunomodulatory agent in dietary supplements.

New Insights into the Structure-Activity Relationship of a Novel Immunomodulatory Peptide (HPHPHLSF) from Casein Hydrolyzed by Kluyveromyces marxianus JY-1: Molecular Docking, Interaction Evaluation, and HOMO Analysis.

Casein is rich in immunomodulatory peptides. In this study, the release of casein-derived immunomodulatory peptides by Kluyveromyces marxianus JY-1 was investigated for the first time, and an immunosuppressive mouse model was used to evaluate the immunomodulatory activity in the casein hydrolysate. The results showed that the cellular and humoral immunity of immunosuppressed mice could be significantly enhanced by casein hydrolysate. Peptide HPHPHLSF with high immunomodulatory activity from casein hydrolysate was screened using the virtual screening technique. HPHPHLSF possessed strong immunomodulatory activity and significantly upregulated the expression of IL-6, IL-1β, and TNF-α. Next, the interaction of HPHPHLSF with TLR2/4 on the cell surface of RAW264.7 cells was further elucidated by molecular docking and combined analysis of double-stranded small interfering RNA and receptor inhibitors. Further, the results of the highest occupied molecular orbital energy distribution elucidated that the histidine active site C48═O49 played an important role in the immunomodulatory activity of HPHPHLSF. This study confirmed that casein hydrolyzed by K. marxianus JY-1 was a natural immunomodulator, while the structure-activity relationship analysis provided new theoretical and technical support for the targeted preparation and screening of casein-derived immunomodulatory peptides.

Supercritical CO2 extracts of propolis inhibits tumor proliferation and Enhances the immunomodulatory activity via activating the TLR4-MAPK/NF-κB signaling pathway

Propolis is a natural immunomodulator with anticancer activity. This study investigated the immunomodulatory mechanism and anti-tumor activity of supercritical CO2 extracts of propolis (SEP) in tumor-bearing immunosuppression mice. We used cyclophosphamide (CTX) to construct the immunosuppressive mice model and then inoculated them with CT26 cells to build the CT26 tumor-bearing immunosuppression mice model. Upon treatment with SEP, tumor proliferation in mice was markedly suppressed, with tumor volumes decreasing from 1881.43 mm3 to 1049.95 mm3 and weights reducing from 2.07 g to 1.13 g. Concurrently, the immune system recovered well, and the spleen and thymus indexes increased significantly. The total T lymphocyte (CD3+ T cell) contents in the spleen and blood recovered from 11.88 % to 21.19 % and 15.32 % to 22.19 %, respectively. In addition, the CD4+ /CD8+ ratio has returned to a healthy level, 3.12 in the spleen and 5.42 in the blood. The levels of IL-1β, IL-6, and TNF-α were increased by 2.17, 2.76, and 7.15 times in the spleen, 2.76, 1.92, and 3.02 times in the serum. Moreover, the western blot results showed that SEP treatment increased the expression of toll-like receptor 4 (TLR4) and the phosphorylation of p38, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p65. These results indicated that SEP activated the immune activity of RAW 264.7 macrophages through the TLR4-mitogen-activated protein kinase (MAPK)/nuclear factor kappa B (NF-κB) signaling pathway to exert immunomodulatory function and inhibit tumor proliferation. This study facilitated the further application of SEP as a potential immunomodulatory and anti-tumor functional food.

Immunomodulatory Activity and Inhibitory Effects of Viscum album on Cancer Cells, Its Safety Profiles and Recent Nanotechnology Development.

Viscum album has been employed traditionally to treat various ailments including as add-on therapy for cancer treatment. V.album formulations have been employed as adjuvants in cancer treatment due to their immunomodulatory activities as well as to alleviate the side effects of conventional cancer therapies. The present review provides updated information from the past 10 years on the immunomodulatory activity and inhibitory effects of V.album on cancer cells, its safety profile, and recent nanotechnology development. V.album extracts and their bioactive phytochemicals, particularly lectins, viscotoxins, and polyphenols, have demonstrated immunomodulatory activity and inhibitory effects against various types of cancer, with low cytotoxicity and side effects, in experimental studies and demonstrated promising anticancer activity in clinical studies in cancer patients. V.album extracts have been shown to enhance immune function by promoting cytokine secretion and inducing both innate and adaptive immune responses, which can help improve immune surveillance against cancer cells. The development of V.album nanoparticles has boosted their biological activities, including inhibitory activity on cancer cells, and could possibly reduce undesired side effects of the plant. Further prospective studies on the plant as a source of new medicinal agents for use as an adjuvant in the treatment of cancer must be performed to provide sufficient efficacy and safety data.

Unlocking the Potential of Food Waste: A Review of Multifunctional Pectins.

This review comprehensively explores the multifunctional applications of pectins derived from food waste and by-products, emphasizing their role as versatile biomaterials in the medical-related sectors. Pectins, known for their polyelectrolytic nature and ability to form hydrogels, influence the chemical composition, sensory properties, and overall acceptability of food and pharmaceutical products. The study presents an in-depth analysis of molecular parameters and structural features of pectins, such as the degree of esterification (DE), monosaccharide composition, galacturonic acid (GalA) content, and relative amounts of homogalacturonan (HG) and rhamnogalacturonan I (RG-I), which are critical for their technofunctional properties and biological activity. Emphasis is placed on pectins obtained from various waste sources, including fruits, vegetables, herbs, and nuts. The review also highlights the importance of structure-function relationships, especially with respect to the interfacial properties and rheological behavior of pectin solutions and gels. Biological applications, including antioxidant, immunomodulatory, anticancer, and antimicrobial activities, are also discussed, positioning pectins as promising biomaterials for various functional and therapeutic applications. Recalled pectins can also support the growth of probiotic bacteria, thus increasing the health benefits of the final product. This detailed review highlights the potential of using pectins from food waste to develop advanced and sustainable biopolymer-based products.

The role of quercetin in NLRP3-associated inflammation.

Quercetin is a natural flavonoid that is widely found in fruits and vegetables. As an important flavonoid, it exhibits a wide range of biological activities, including antioxidant, anti-inflammatory, antiviral, immunomodulatory, and analgesic activities. Quercetin exerts powerful antioxidant activity by regulating glutathione, enzyme activity, and the production of reactive oxygen species (ROS). Quercetin exerts powerful anti-inflammatory effects by acting on the Nod-likereceptorprotein3 (NLRP3) inflammasome. In diabetes, quercetin has been shown to improve insulin sensitivity and reduce high blood sugar level, while, in neurological diseases, it potentially prevents neuronal degeneration and cognitive decline by regulating neuroinflammation. In addition, in liver diseases, quercetin may improve liver inflammation and fibrosis by regulating the NLRP3 activity. In addition, quercetin may improve inflammation in other diseases based on the NLRP3 inflammasome. With this background, in this review, we have discussed the progress in the study on the mechanism of quercetin toward improving inflammation via NLRP3 inflammasome in the past decade. In addition, from the perspective of quercetin glycoside derivatives, the anti-inflammatory mechanism of hyperoside, rutin, and isoquercetin based on NLRP3 inflammasome has been discussed. Moreover, we have discussed the pharmacokinetics of quercetin and its nanoformulation application, with the aim to provide new ideas for further research on the anti-inflammatory effect of quercetin and its glycoside derivatives based on NLRP3 inflammasome, as well as in drug development and application.

Peronema canescens as a Source of Immunomodulatory Agents: A New Opportunity and Perspective.

Immunomodulators are pivotal in managing various health conditions by regulating the immune response by either enhancing or suppressing it to maintain homeostasis. The growing interest in natural sources of immunomodulatory agents has spurred the investigation of numerous medicinal plants, including Peronema canescens, commonly known in Asia as sungkai. Traditionally used for its medicinal properties in Southeast Asia, Peronema canescens belongs to the Verbenaceae family and has garnered significant attention. This review discusses the immunomodulatory activity of the active compounds in Peronema canescens and explores the potential directions for future research.

Immunoprognostic analysis of indoleamine 2,3-dioxygenase 1 in patients with cervical cancer.

The incidence of cervical cancer is increasing. Immunotherapies show better patient outcomes than monotherapies; however, the mainstay treatment for cervical cancer remains surgery and chemotherapy. Indoleamine 2,3-dioxygenase 1 (IDO1) acts on multiple tryptophan substrates, exhibiting antitumor, immunomodulatory, and antioxidant activities. Despite the association of elevated IDO1 expression with unfavorable outcomes in various cancers, its precise function in cervical cancer remains ambiguous. Here, we explored the prognostic significance of IDO1 in cervical carcinoma. Gene expression datasets were obtained from The Cancer Genome Atlas. Gene Expression Omnibus datasets were used for differential expression and functional correlation analyses. Using Human Protein Atlas alongside Tumor-Immune System Interaction Database, we assessed the association of IDO1 with survival rates. Given the link between cervical cancer prognosis and immune invasion, CIBERSORT was used to assess the connection between immune cells and IDO1, while the percentage of tumor-penetrating immune cells based on IDO1 expression in cervical cancer patients was analyzed using Tumor-Immune System Interaction Database. Incorporating a clinicopathological characteristic-based risk score model with IDO1 risk score, we devised a nomogram to predict cervical cancer patient survival. The effects of IDO1 in immune regulation and its prognostic significance were validated using data from patients with cervical cancer obtained from The Cancer Imaging Archive database. Compared with that in normal cervical tissues, IDO1 expression was significantly upregulated in cervical cancer tissues and significantly correlated with cervical cancer progression and prognosis. IDO1 expression showed a positive association with monocyte and macrophage abundance, while exhibiting a negative correlation with that of endothelial cells and eosinophils. Cox regression analyses highlighted IDO1 as the core immune gene implicated in cervical cancer. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed an association of IDO1 with the metabolic pathways of tryptophan, phenylalanine, and tyrosine. Univariate and multivariate analyses revealed that elevated IDO1 expression correlates markedly with cervical cancer outcomes, suggesting it as a promising therapeutic target. The Cancer Imaging Archive data analysis revealed that the impact of anti-PD1 and CTLA4 therapy is more pronounced in cervical cancer patients exhibiting elevated IDO1 expression. IDO1 is a potential target for immunotherapy for cervical cancer.

Functional Characterization of an Arylsulfonamide-Based Small-Molecule Inhibitor of the NLRP3 Inflammasome.

Considerable evidence indicates that the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome plays key roles in human pathophysiology, suggesting it as a potential drug target. Currently, studies have yet to develop compounds that are promising therapeutics in the clinic by targeting the NLRP3 inflammasome. Herein, we aim to further biologically characterize a previously identified small-molecule inhibitor of the NLRP3 inflammasome from our group, YM-I-26, to confirm its functional activities. We showed that YM-I-26 is highly selective toward the NLRP3 inflammasome and binds to NLRP3 directly. A systemic analysis revealed YM-I-26 with inflammation-related and immunomodulatory activities by the Eurofins BioMAP Diversity PLUS panel. In addition, studies using the mouse microglia BV2 cell model demonstrated that YM-I-26 is not cytotoxic, improved the phagocytotic functions of BV2 cells toward beta-amyloid, and suppressed the production of cytokines of IL-1β and IL-10 upon the activation of the NLRP3 inflammasome. Collectively, our studies support the functional activities of YM-I-26 as a NLRP3 inhibitor in physiologically relevant cell models, and warrant future studies of YM-I-26 and its analogs to advance the drug development as potential therapeutics.

Impact of neutrophil‐activating protein conservation on diagnostic tests and vaccine design

AbstractBACKGROUNDThe neutrophil activating protein (NAP) is a highly immunogenic and virulence factor of Helicobacter pylori, presenting inflammatory and immunomodulatory activity. Consequently, NAP has been explored as a diagnostic and therapeutic target. However, when evaluating a target protein to design diagnostic methods or vaccines, it is critical to determine the protein conservation among the bacterial population, as well the impact of alterations of amino acid residues on the protein antigenic profile.RESULTSIn the present work, NAP conservation and theoretical antigenicity were determined among 51 sequences from H. pylori isolated from patients worldwide. A high NAP conservation (83%) was observed, where 17 amino acid residues, among the 144 residues of the protein, were polymorphic. Alterations at these polymorphic sites had a theoretically low impact on predicted antigenicity, where only 5 NAPs out of 51 NAPs presented a slightly different antigenic profile in relation to the consensus sequence. According to that, it was possible to recognize in western blotting 93% of NAP from different bacteria (n = 15) using polyclonal antibodies developed against a specific NAP.CONCLUSIONSIt was predicted that when working with polyclonal antibodies or large NAP fragments for diagnostic and vaccine design, slight variation in protein sequence will have a minimal impact on NAP recognition. However, if a NAP monoclonal antibody or small NAP epitopes are considered, it is critical to select the most conserved and antigenic NAP regions, to maximize the coverage of NAP variants. © 2024 Society of Chemical Industry (SCI).

Characterization of salt brine sulfated polysaccharides: immunomodulatory activity based on gut microbiota

A sulfated polysaccharide consisting of two components with molecular weights of 439 kDa and 16 kDa was extracted from the salt brine. The structural properties, immunomodulatory activity, in vitro fermentation behaviors, and effects of SP on regulating the gut microbiota were investigated. The chemical composition and monosaccharide composition analysis showed that the neutral sugar, protein, uronic acid, and sulfated group contents of SP were 60.42 ± 0.04%, 2.90 ± 0.01%, 13.34 ± 0.01% and 10.51 ± 0.01%, respectively, containing arabinose, galactose, glucose, rhamnose, xylose, mannose, and glucuronic acid in a molar ratio of 33.24:19.18:16.64:13.25:8.31:4.11:5.27. Results from the macrophage cell model showed that SP intervention improved the proliferation activity, phagocytosis of neutral red, and production of IL-6 and TNF-α in RAW 264.7. Furthermore, in vitro fermentation of SP by gut microbiota showed that SCFA production in all treatment groups was significantly higher than that of the blank control group after 48 h of fermentation, especially butyric acid which was 1.70 folds that of the control group. Moreover, long-term fermentation (48 h) of SP improved the diversity of microbiota, decreased the F/B ratio (30.75 at 0 h vs. 1.22 at 48 h), and promoted the growth of probiotics (Parabacteroides, Bacteroidetes, Ruminococcaceae, and Phascolarctobacterium). The positive regulatory effect of SP on the gut microbiota and its metabolites is considered a potential target for its immunomodulatory activity.Graphical abstract

Immunomodulatory activity of red algal galactans and their partially depolymerized derivatives in RAW264.7 macrophages

Funoran and furcellaran were isolated through a successive cascade extraction process, followed by the depolymerization of extracted polymers via an auto-hydrolysis process. The molecular weight and structural peculiarities of both native and partially depolymerized polysaccharides were investigated using size-exclusion chromatography (SEC), FTIR, and NMR spectroscopy. Immunotropic effects of the native and partially depolymerized polysaccharides were explored through various in vitro assays. Although both higher and lower molecular weight funoran exhibited anti-inflammatory activity on LPS-stimulated RAW264.7 cells by significantly downregulating iNOS and COX-2 gene expression, as well as the secretion of pro-inflammatory cytokines, native funoran performed slightly better. Conversely, higher molecular weight furcellaran remarkably activated RAW264.7 cells compared to the non-treated control by inducing inflammatory mediators and pro-inflammatory cytokines, including the anti-inflammatory cytokine IL-10. Lower molecular weight furcellaran was unable to activate the macrophages, showing a similar behavior pattern to funoran samples in LPS-treated cells. Furthermore, the TLR4/NF-κB signaling pathway appears to be modulated by sulfated polysaccharides, leading to both anti-inflammatory and immunostimulatory responses in RAW264.7 cells through blocking and activating mechanisms. These findings indicate that sulfated polysaccharides could be promising therapeutic agents, and indeed, the molecular weight of polysaccharides plays a crucial role in the immune response of RAW264.7 macrophages.

Soluble PD-L1 shows no association to relapse and overall survival in early stage non-small cell lung cancer (NSCLC)

BackgroundCancer immune evasion is critical in non-small cell lung cancer (NSCLC) and has been targeted by immunotherapy. High soluble (s)PD-L1 is associated with reduced survival and treatment failure in advanced stages. Here we evaluated the effects of sPD-L1 on T cells, relapse free survival, and overall survival in early stage NSCLC. MethodsIn vitro T cell stimulation was performed in the presence of sPD-L1 to evaluate its immunomodulatory activity. Data from The Cancer Genome Atlas (TCGA) were investigated for PD-L1 splice variants and enzymes involved in proteolytic cleavage (i.e. ADAM10). Plasma from 74 NSCLC (stage IA-IIIB), as well as an additional 73 (control cohort) patients was collected prior to curative surgery. Thereafter sPD-L1 levels from an immunosorbent assay were correlated with patient outcome. ResultsIn vitro sPD-L1 inhibited IFN-γ production and proliferation of T cells and induced a terminal effector CD4 T cell subtype expressing CD27. Data from the TCGA demonstrated that elevated mRNA levels of ADAM10 is a negative predictor of outcome in NSCLC patients. To investigate the clinical relevance of these in vitro and TCGA findings, we quantified sPD-L1 in the plasma of early-stage NSCLC patients. In the first cohort we found significantly higher sPD-L1 levels in relapsing NSCLC patients, with a multivariate analysis revealing high sPD-L1 (>1000 pg/mL) as an independent predictor of survival. However, these findings could not be validated in two independent control cohorts. DiscussionAlthough in vitro and TCGA data support the suppressive effect of sPD-L1 we were unable to translate this in our clinical setting. These results may be due to the small patient number and their heterogeneity as well as the lack of a standardized sPD-L1 ELISA. Our inconclusive results regarding the value of sPD-L1 in early stage NSCLC warrant assay validation and further investigation in larger (neo-)adjuvant trials.

Colon Cancer Cells Treated with Lacticaseibacillus casei Undergo Apoptosis and Release DAMPs Indicative of Immunogenic Cell Death.

Probiotic bacteria, and especially lactic acid bacteria, have long been known to wield a variety of health-beneficial effects, including antioxidant, antimicrobial, anti-inflammatory, immunomodulatory, and anticancer activities. However, our understanding of the mechanisms involved in these activities remains incomplete. In this study, we wished to investigate the processes that give rise to the anticancer activity of Lacticaseibacillus casei ATCC393 and the possibility that immunogenic cell death of cancer cells can be induced following treatment with this probiotic. In both cell lines that we have examined, we detected notable pro-apoptotic signaling, including the upregulation of death receptors, that culminated in the activation of caspase 3, the endpoint and most characteristic effector molecule of all pro-apoptotic cascades. In addition, we identified damage-associated molecular patterns associated with immunogenic cell death. Calreticulin exposure on the outer cell membrane, HMGB1 translocation outside the nucleus and depletion of intracellular ATP was evident in both cancer cell lines treated with the probiotic, while expression of type I interferons was upregulated in CT26 cells. Our findings suggest that treatment with the probiotic induced apoptosis in cancer cells, mediated by extrinsic death receptor signaling. Moreover, it resulted in the release of molecular signals related with immunogenic cell death and induction of cancer cell-specific adaptive immune responses.

Purified L-glutaminase effects against multidrug-resistant Pseudomonas aeruginosa in experimental vaginosis model: An immunological and histopathological observation

The antimicrobial activity of crude and purified L-glutaminase (EC 3.5.1.2), obtained from Lactobacillus gasseri, was evaluated against multidrug-resistant Pseudomonas aeruginosa in the in vivo vaginosis condition. The L-glutaminase possessed significant antimicrobial and anti-biofilm formation activity against multi-drug resistance P. aeruginosa, which were confirmed in the BALBc rat vaginosis model, together with its effects on the immunological and histopathological aspects. The untreated animals showed heavy vaginitis, characterized by sub-epithelial edema and infiltration of mononuclear leukocytes, perivascular heavy inflammatory cells infiltration in the vaginal tissue, and moderate stromal edema. However, the L-glutaminase treatment exhibited no changes in vaginal tissue structure with normal appearance of the epithelium and lamina propria with marked repair of the vaginal section when compared with normal, uninfected, control group A. The immunomodulatory actions of the L-glutaminase were confirmed by observance of higher concentrations of tumor necrosis factor-γ (TNF-γ), and interleukin −12 (IL-12) in treated animals, while the interleukin-10 (IL-10) was higher in the infected, untreated animals' sera samples. Therefore, the L-glutaminase showed corrective and healing actions, which were observed through histopathological observations of the vaginal tissue. The investigations led to imply that L-glutaminase may have the potential to be an effective antimicrobial agent for preventing and inhibiting bacterial growth, as well as inhibiting the biofilm formation in the P. aeruginosa-originated vaginosis. The observations may be of promising value for future clinical use.

Anti-Inflammatory And Immunomodulating Activity of the 1(10) Β-Epoxy-5,7α,6β(Н)-Guai-3(4),11(13)-Dien-6,12-Olide

BACKGROUND: The article discusses the results of studies of the anti-inflammatory and immunomodulatory activity of the sesquiterpene lactone 1(10)β-epoxy-5,7α,6β(Н)-guai-3(4),11(13)-diene-6,12-olide (hereinafter referred to as epoxyguaianolide). AIM: The article discusses the results of studies of the anti-inflammatory and immunomodulatory activity of the sesquiterpene lactone 1(10)β-epoxy-5,7α,6β(Н)-guai-3(4),11(13)-diene-6,12-olide. MATERIALS AND METHODS: A non-steroidal anti-inflammatory drug diclofenac sodium at a dose of 25 mg/kg (ampoules, 2 mL each, manufactured by Germany) was used as a reference drug in models of acute aseptic inflammation and cotton ball granuloma. On the model of cyclophosphamide immunosuppression, the immunomodulator levamisole at a dose of 3 mg/kg, was used as a reference drug. The immunomodulating effect of the test samples was studied on outbred adult male rats and outbred mice. We calculated the arithmetic mean, the mean square error of the arithmetic mean, the significance of the difference between the mean Рt according to the student’s t-test (TTEST), and the significance of the difference in Pf variances according to the Fisher f-test. RESULTS: The results of the experiments indicate the presence of a pronounced immunomodulatory activity in epoxyguaianolide. When interacting with cells of the immune system, epoxyguaianolide activated the studied parameters in animals with immunosuppression, whereas in intact animals, it did not affect the above parameters. On the model of cyclophosphamide immunosuppression, it was found that epoxyguaianolide has a pronounced immunomodulatory effect, manifested in an increase in the number of T-lymphocytes and a subpopulation of theophylline-resistant T-lymphocytes, an increase in the number of theophylline-sensitive T- and B-lymphocytes, activation of phagocytosis processes, and increased cell migration. This action of epoxyguaianolide eliminates the minimal possibility of hyperactivation of the immune system, which is an important condition for its use as an immunotropic agent. CONCLUSIONS: The conducted experiments made it possible to recommend epoxyguaianolide for clinical trials as an immunomodulating agent in the treatment of chronic inflammatory diseases accompanied by secondary immunodeficiencies.

Viola stocksii: A rich source of antioxidant and anti-inflammatory flavonoid glycosides with converged therapeutic potential against SARS-CoV-2 MPro, spike trimer, and surface glycoproteins

This research presents the first comprehensive investigation into the phytochemical and nutraceutical properties of Viola stocksii. The study aimed to identify the therapeutic efficacy and bioactive constituents of this medicinal herb, traditionally used as an herbal tea for managing respiratory and abdominal issues in remote areas of Pakistan. The therapeutic efficacy of the plant was assessed as an anti-oxidant-rich health tonic coupled with anti-inflammatory and immunomodulatory activities against COVID-19 and its evolving variants. The study involved sequential extraction, bio-assay guided fractionation, isolation, characterization, and biological evaluation. Preliminary in vitro studies have demonstrated that polar fractions of the plant possess significant antioxidant and antiinflammatory potential. A diverse combination of chromatographic techniques (Diaion resin HP-20, Sephadex LH-20, ODS-C18, and RP-HPLC) facilitated the purification of active constituents of the plant. Comprehensive analysis of NMR (1H, 13C, and 2D NMR) and mass (EIMS, HR-EIMS, and FAB-HRMS) spectral data lead to characterize the isolated compounds (1–4) as flavonoid glycosides. Results from in vitro and in-silico evaluations revealed that the isolated glycosides exhibit high antioxidant, antiinflammatory, and immunomodulatory activities, which synergistically show promising potential in combating SARS-CoV-2 variants. Besides MPro, therapeutic potential against two unexplored proteins i.e. the spike trimer (7WZ1) and surface glycoproteins (7QTJ) was studied for the first time. Kaempferol (KamGluαRh) and quercetin (QurGluαRh) carrying rutinoside, were identified as the most active constituents of the plant, with the highest binding affinities in the range of −10 to −13 kcal/mol. Ten distinct active regions were explored on the spike trimer, including strong binding affinity with the essential RBD region, spike head, and tail of the protein. MD simulation validated the stability of Mpro-KamGluαRh and Mpro-QurGluαRh complexes evidenced by an average RMSD of 0.2 nm led to the mechanistic understanding of Mpro inhibition. ADMET and pharmacokinetic studies authenticated the therapeutic ability and drug-likeness profiling of all extracted compounds. Results revealed the beneficial role of glycosides in boosting the medicinal effectiveness of flavonoids and the potential of Viola stocksii as an immunity booster against respiratory infections like COVID-19.

Intestinal probiotic-based nanoparticles for cytotoxic siRNA delivery in immunotherapy against cancer

Immunogene therapy has emerged as strategy against cancer by introducing immune-stimulating components into gene therapy. However, there is still a need for an ideal platform to achieve both immune stimulation and efficient gene delivery. Lactobacillus reuteri has potential immunomodulatory activity owing to its unique antigenicity, which is potentially relevant to cancer progression. Here, we designed a novel non-viral siRNA vector (DMPLAC) by encapsulating Lactobacillus reuteri lysate in DMP. DMPLAC can promote maturation and activation of immune cells, increase infiltration of APC and cytotoxic T cells in tumor microenvironment, and exhibit tumor suppressive effects. Loading of siRNA targeting Stat3, DMPLAC/siStat3 further inhibits tumor in multiple models. We designed a strategy that combines immune activation with Stat3 silencing, triggering an immune response and tumor killing. This dual-functional design provides a new choice in development of effective immunogene therapy.