Search Results (228)

Background: Tuber indicum is a rare edible and medicinal ectomycorrhizal fungus, and the polysaccharide fractions have attracted extensive research attention owing to their potent immunomodulatory potential. Objective/Methods: To systematically characterize the structural features and gut microbiota-mediated immunoregulatory mechanisms of water-extracted polysaccharide (TIWP) and alkali-extracted polysaccharide (TIAP) from T. indicum, two polysaccharide fractions were prepared and comprehensively structurally characterized in this study. Results: The maximum molecular weight of TIWP reached 2.65 × 10⁷ Da, which was dominated by glycosidic linkages of →3)-Glcp-(1→, →2,4)-Glcp-(1→ and →3,6)-Glcp-(1→. TIAP possessed a higher molecular weight up to 3.87 × 10⁷ Da, with predominant glycosidic bonds of →3,6)-Glcp-(1→, →4)-Glcp-(1→ and →2,4)-Glcp-(1→. In vivo bioactivity evaluation results demonstrated that both TIWP and TIAP significantly restored the proportions of CD3⁺ T and CD19⁺ B lymphocyte subsets in peripheral blood and spleen, and alleviated pathological damage in splenic and colonic tissues. Distinct regulatory patterns of gut microbiota were observed between the two polysaccharides: TIWP markedly enriched the genera Lactobacillus and Ruminiclostridium, whereas TIAP elevated the relative abundances of Lactobacillus and Alloprevotella. Untargeted metabolomics combined with KEGG pathway enrichment analysis revealed that TIWP and TIAP activated the pantothenic acid and coenzyme A biosynthesis pathway and the linoleic acid metabolism pathway. Conclusions: Collectively, TIWP and TIAP alleviated CTX-triggered immunosuppression through distinct “gut microbiota-metabolite-immunity” regulatory networks due to their structural disparities. This study provides a theoretical basis for the development of gut microecology-targeted functional foods with immunomodulatory functions. Full article

Background: Non-small-cell lung cancer (NSCLC) is the leading cause of cancer-related mortality worldwide. Although immune checkpoint inhibitors (ICIs) have significantly improved clinical outcomes, most patients do not respond to treatment or fail to achieve durable responses. Histone deacetylase inhibitors (HDACi) have emerged as promising immunomodulatory agents, with the potential to sensitize tumors to ICIs. We investigated the immunomodulatory effects of entinostat, a class I HDACi, in combination with dual ICI (anti-PD-1 and anti-CTLA-4) as well as with T-cell receptor (TCR) engineered T cells in preclinical NSCLC models. Methods: We employed human NSCLC cell lines and the immunogenic KRASG12D/p53-mutant KPN1.1 murine NSCLC cell line. In vitro, we assessed entinostat-induced changes in MHC class I and PD-L1 expression. In addition, we evaluated the effects of entinostat on a KRASG12D-specific TCR. In vivo, therapeutic efficacy and immune modulation were assessed by transplanting KPN1.1 cells subcutaneously and orthotopically into immunocompetent mice, followed by treatment with dual ICI, with or without entinostat. Immune populations in the spleen and blood were subsequently analyzed. Results: In vitro, entinostat induced the upregulation of MHC-I and PD-L1 expression in both human and murine NSCLC cell lines. In addition, entinostat treatment significantly enhanced antigen-specific tumor recognition and killing by T cells engineered to express a KRASG12D-specific TCR. In vivo, the addition of entinostat to dual immune checkpoint inhibition showed an incremental trend toward improved tumor growth control. Notably, entinostat plus dual ICI enhanced systemic immune activation, increasing circulating and splenic T-cell populations and significantly expanding both antigen-specific and overall effector CD8⁺ T cells. Consistently, the ex vivo co-culture of splenocytes from KPN1.1-bearing mice with KPN1.1 tumor cells demonstrated enhanced CD8⁺ antigen-specific T-cell recognition. Conclusions: In human and murine NSCLC models, entinostat potentiates TCR- and ICI-mediated tumor recognition through tumor-intrinsic and systemic immune modulation. These effects were reflected by increased MHC-I expression, expansion of antigen-specific effector CD8⁺ T cells, and enhanced CD8⁺ T-cell tumor recognition. These findings support a further evaluation of entinostat as a strategy to improve immunotherapy efficacy in NSCLC. Full article

Onion peel extract (OPE) is rich in polyphenolic compounds with antimicrobial potential. Salmonella Enteritidis (SE) infection in young broiler chicks causes morbidity, reduced growth, and contributes to human gastroenteritis through contaminated poultry products. The spleen is a key secondary lymphoid organ coordinating systemic responses to pathogens in chicken. This study evaluated how dietary OPE influences spleen metabolic profiles during SE infection. Day-old Ross 708 male chicks (n = 128) were assigned to four treatments: CON, CON-SE, OPE (6 g/kg), and OPE-SE. Chicks in CON and OPE received sterile broth, whereas CON-SE and OPE-SE received 2.25 × 10⁸ CFU/mL SE at 2 d of age. At 5 and 12 dpi, spleens from six chicks per treatment were collected for untargeted HPLC-MS metabolomics. A total of 857 metabolites were identified and analyzed using MetaboAnalyst 6.0 (p < 0.05; fold change ≥ 2.0; VIP score > 1.0). In CON-SE chicks, energy generating metabolites (6-phosphogluconic acid, methylmalonic acid, propionic acid) increased, while 13,14-dihydro-15-keto-prostaglandin D2 and kynurenic acid decreased. Dietary OPE elevated several dipeptides (L-Val-Gly, L-Leu-Gly, Gly-Gly-Leu, L-Val-L-Met) and reduced ATP linked metabolites (3,6-di-O-methyl-beta-D-glucose and 3-O-beta-D-galactosyl-sn-glycerol). Enrichment analysis showed that SE infection altered valine, leucine, and isoleucine degradation and aromatic amino acid biosynthesis, whereas OPE enriched galactose and biotin metabolism in uninfected chicks, but enriched tryptophan, taurine and hypotaurine metabolism in SE-infected chicks. Overall, dietary OPE optimized response of metabolic pathways associated with immune activation, unlike corresponding pathways in CON-SE birds. Full article

The current use of artificial light during the natural dark phase has acquired contaminant dimensions, known as “light pollution”. It is well known that exposure to dim light at night (dLAN) during the postnatal period severely impairs the immune system and related organs, but few reports have demonstrated the effects of dLAN during the fetal period. This study, therefore, examines whether exposure to dim light at night during two critical developmental windows (i.e., prenatal and postnatal periods) leads to long-lasting dysregulation of circadian, behavioral, and immune organization, as well as spleen immune responses, in early adulthood. To address this question, these outcomes were assessed using two defined sampling time points. To answer this question, we exposed two groups of C57BL/6J male mice to dim night light during the gestational and postnatal periods and compared them with control groups that were exposed to light–dark conditions (12 h each, LD). Parametric and non-parametric activity/rest values were analyzed with circular statistics. Compared to their controls, we found differences in alpha, onset, offset, M10, and L5 start time in dLAN groups. We also assessed the transcript levels of clock genes and inflammatory mediators in spleen tissue and found a dampening of daily variation in mRNA expression in both experimental groups. Finally, we used an ovalbumin (OVA) allergy challenge to test the B-cell response in the spleen and found a significantly higher cell recruitment to the spleen and more anti-OVA IgE. Together, these results clearly show that dLAN, at two ZT sampling points, affects peripheral molecular clocks and responses in the spleen, and that these effects are independent of the life stage at which exposure to dim light at night occurs. Full article

The Syrian golden hamster (Mesocricetus auratus) is a universally accepted model for visceral leishmaniasis (VL) due to its ability to mimic human disease pathology. Mus musculus (BALB/c) is preferred for evaluating pharmaceutical and immunological responses. This study focuses on the precise role of gene signatures in L. donovani-infected M. auratus and M. musculus, using transcriptomic analysis. Principal component analysis (PCA) revealed distinct clustering among the four groups (uninfected vs. infected spleen samples from M. auratus and M. musculus). After differential expression analysis, 2054 genes in M. auratus and 1108 in M. musculus were found to be differentially expressed, with 153 genes common to both species. Except for 31 genes, most of the commonly dysregulated genes show a similar expression pattern. Although Th1-mediated immune signaling was observed in both cases, the overexpression of LAG3 in both infected groups underscores the important role of T cell exhaustion. Immunological responses against parasite infection in M. auratus appear to be more aggressive, while M. musculus seems more intense. Interestingly, only the M. musculus-infected group shows overexpression of IL-10. Without a definitive role for IL-10, the overexpression of Tgm2, Clec7a, and Adora2b in both species may drive disease outcome. These findings elucidate the immunological mechanisms driving the pathogenesis of VL in rodent models. Full article

Background/Objectives: Chronic pain is a significant clinical challenge, in part due to the absence of reliable objective biomarkers for its evaluation and treatment. Growing evidence indicates that immune cells, including natural killer (NK) cells, are involved in the regulation of pain processes. NK cells are innate cytotoxic lymphocytes whose functional status may mirror underlying pathological pain states. In this study, we investigated μ-opioid receptor (MOR) expression and functional alterations of NK cells in a murine model of neuropathic pain induced by chronic constriction injury (CCI). Methods: Mice were divided into three groups: Sham (sciatic nerve exposure without ligation), CCI 14-day, and CCI 21-day groups. At the respective time points, animals were sacrificed and spleens were collected for analysis. Splenocytes were isolated by mechanical dissociation followed by centrifugation and erythrocyte lysis. Lymphocytes were analyzed by flow cytometry to evaluate MOR expression in NK cells and their degranulation activity (CD107a assay). Cells were incubated with fluorochrome-conjugated antibodies against NK cell markers (NK1.1, CD3, Ly49A, Ly49C/I) in combination with anti-MOR and anti-Interferon γ antibody (IFN-γ). Immunofluorescence and confocal microscopy analyses were performed to assess MOR localization and granzyme localization, supporting CD107a-mediated degranulation. Results: Flow cytometry analysis revealed a significant reduction in surface MOR expression on total NK cells from CCI mice compared with sham controls at 14 and 21 days post-injury, a finding corroborated by immunofluorescence evidence of MOR cellular internalization. Functionally, CCI induced a marked decrease in CD107a expression and impaired IFN-γ production both under basal conditions and following PMA/ionomycin stimulation, indicating a hyporesponsive state of NK cells. Consistently, confocal microscopy revealed extracellular release of Granzyme A following CCI, suggesting dysregulated degranulation. Conclusions: Neuropathic pain is associated with a remodeling of NK cell phenotype and effector functions, characterized by impaired cytotoxic activity and cytokine production, along with modulation of inhibitory receptor expression. Notably, MOR-reduced surface expression in NK cells emerges as a potential biomarker of neuropathic pain. Further studies are needed to elucidate the molecular mechanisms regulating MOR expression and its relationship with NK cell hyporesponsiveness and degranulation in chronic pain conditions. Full article

Background/Objectives: Immunodeficiency can be induced by a variety of factors, such as aging, stress and poor nutrition, and leads to increased susceptibility to infection and disease. The current research was conducted to determine the immunoenhancing potential of yam and its underlying mechanism in a murine model of cyclophosphamide (CTX)-induced immunosuppression. Methods: The gut microbial community and generation of short-chain fatty acids (SCFAs) in response to yam were analyzed by 16S rRNA sequencing and GC-MS. The immune cells in the spleen were analyzed using flow cytometry. GPR41/GPR43/GPR109A triple-knockout mice were used to demonstrate the critical involvement of SCFAs in mediating the protective effect of yam, and RNA-sequencing technology was applied to investigate the potential mechanism by which yam orchestrated the observed metabolic, immune and reparative responses. Results: Yam alleviated the decline in spleen and thymus indices and modulated the frequency of B cells and CD4⁺ and CD8⁺ T cells and promoted the production of IgA, IgG and IgM. Yam increased the secretion of cytokines in the intestine and upregulated the levels of claudin and ZO-1. Yam also increased the content of SCFAs and induced beneficial modifications to the gut microbiota composition. The immune-enhancing activity of yam was confirmed, as evidenced by a notable decrease in viral load in immunosuppressed mice inoculated with influenza virus and its capacity to mitigate inflammatory response in pulmonary tissues. Conclusions: This study suggests that yam enhances immunity by synergistically regulating the gut–immune axis, supporting its development as a functional food intervention in managing immunodeficiency conditions. Full article

Background/Objectives: Immune checkpoint inhibitors targeting the PD-1/PD-L1 axis have transformed cancer treatment, yet therapeutic responses remain limited in many solid tumors due to poor and uneven drug distribution within the tumor microenvironment (TME). Here, we evaluated whether co-formulation of an anti-PD-1 antibody (RMP1-14, murine surrogate for pembrolizumab) with Imagent^® microbubble/liposome (MBLP) complexes and ultrasound activation could enhance tumor-specific delivery while reducing systemic exposure. Methods: Immunocompetent MC-38 colorectal tumor-bearing mice (B6(Cg)-Tyrc-2J/J, 7-week-old females) received isotype control, isotype/MBLP/US, RMP1-14 alone, RMP1-14/MBLP, or RMP1-14/MBLP/US. Survival was analyzed by Kaplan–Meier curves, tumor necrosis by H&E staining, antibody biodistribution by immunohistochemistry, and tumor perfusion by laser speckle imaging. Results: No significant differences in tumor size or body weight were observed between groups. Survival analysis showed significant improvements in the RMP1-14 (p = 0.013) and RMP1-14/MBLP/US (p = 0.047) groups versus isotype controls, with the RMP1-14/MBLP/US group achieving the longest mean survival (57.8 days vs. 26.5 days for RMP1-14 alone) and complete tumor regression in 2/8 mice. The RMP1-14/MBLP/US group demonstrated significantly greater tumor necrosis than all other groups. Immunohistochemical analysis confirmed a 6.1-fold increase in intratumoral antibody accumulation with MBLP/US versus RMP1-14 alone (p = 0.0003), alongside significantly reduced off-target exposure in spleen, liver, kidney, and heart. Laser speckle imaging revealed a transient ~30% increase in tumor perfusion during MBLP/US treatment, consistent with cavitation-mediated hemodynamic effects. Conclusions: These findings demonstrate that MBLP/US co-formulation enhances intratumoral delivery of checkpoint inhibitors, improves survival, and reduces systemic organ exposure, representing a promising platform to improve the efficacy and safety profile of antibody-based immunotherapy. Full article

Feed additives play a crucial role in boosting livestock immunity, but their underlying molecular mechanisms are often incompletely understood. This study used integrated proteomics and metabolomics to systematically investigate the immunomodulatory effects of dietary fermented onion (FO) on the spleen of Liangshan Black Sheep. Results showed that FO supplementation significantly improved systemic antioxidant capacity and immune function, indicated by a higher spleen index and increased serum concentrations of SOD, MCP-1, and IL-2 (p < 0.05). Multi-omics profiling of spleen tissues identified 169 differentially expressed proteins and 168 differential metabolites. Integrated pathway enrichment revealed calcium signaling and purine metabolism as potential core regulatory hubs for the observed immunomodulation. This molecular remodeling correlated with key molecules including protein F2R and metabolites adenosine and hypoxanthine, which may form a coordinated regulatory network. Overall, our findings suggest potential pathways linking dietary FO supplementation to enhanced splenic immune function in Liangshan Black Sheep, likely via synergistic regulation of specific signaling pathways. This work supports FO as a promising functional feed additive and provides a molecular framework for developing novel immunomodulatory strategies in livestock production. Full article

Although immunotherapy has shown great promise in treating various types of cancer, advanced tumors are often refractory due to a highly immunosuppressive tumor microenvironment (TME). We previously engineered a cancer therapeutic vaccine platform, µGCVax, by co-loading tumor antigen peptides, STING and TLR9 agonists into porous silicon microparticles. While effective in models with lower disease burden, its efficacy against advanced colorectal cancer (CRC) was less promising due to the accumulation of myeloid-derived suppressor cells (MDSCs) in TMEs. In this study, we investigated whether µGCVax-based immunotherapy in advanced CRCs could be potentiated via regulating MDSCs to reprogram the TME. In an advanced CT26 murine CRC model, we assessed µGCVax in combination with oxaliplatin, a standard CRC chemotherapeutic with established immunomodulatory effects. We demonstrated that oxaliplatin was preferentially taken up by monocytic MDSCs (M-MDSCs) and effectively reduced their abundance in the bone marrow, blood, spleen, and tumor. Relief of this immunosuppressive TME increased intratumoral infiltration of antigen-specific CD8⁺ T cells. Ultimately, the combination of oxaliplatin with µGCVax induced robust regression of established CRC tumors. These findings highlight that oxaliplatin synergizes with µGCVax by overcoming MDSC-mediated immunosuppression and enhancing antitumor immunity, representing a promising chemo-immunotherapy strategy for advanced CRC. Full article

Background/Objectives: The IUCN endangered spotted pond turtle (Geoclemys hamiltonii) demonstrates markedly reduced resistance to Aeromonas hydrophila-induced infections compared to the red-eared slider (Trachemys scripta). This study investigates the immunological basis for this disparity by analyzing infection outcomes and splenic transcriptomes of both species post-pathogen challenge. Methods: In a preliminary experiment, 32 turtles (16 G. hamiltonii and 16 T. scripta) were exposed to A. hydrophila. Results: G. hamiltonii developed skin ulcer syndrome at a significantly higher incidence (81.25%) than T. scripta (12.5%) (p < 0.05). Comparative transcriptomic analysis identified 19 differentially expressed immune-related genes, with qPCR validation across five tissues (heart, liver, spleen, intestine, blood) revealing pronounced interspecies differences in IL18, STAT1, IFIH1, and CD28 expression. Notably, IL18 and its downstream effector STAT1 were robustly upregulated in T. scripta but were considerably lower in G. hamiltonii, correlating with delayed IFN-γ pathway activation and impaired epidermal barrier repair. Concurrently, CD28 upregulation in T. scripta facilitated rapid T-cell-mediated pathogen clearance, whereas its delayed induction in G. hamiltonii hindered adaptive immunity. These findings implicate dysregulated innate (IL18/STAT1) and adaptive (CD28) immune pathways as key determinants of G. hamiltonii’s susceptibility to bacterial infection. Conclusions: Despite the critical conservation status of G. hamiltonii, the immunological basis underlying its heightened susceptibility to bacterial infections remains largely unexplored; this study addresses this gap by comparing the splenic transcriptomes of G. hamiltonii and T. scripta following A. hydrophila challenge, identifying the dysregulated IL18-STAT1 Immune Axis and CD28-mediated adaptive immunity as key determinants, thereby providing actionable immune targets for conservation breeding and susceptibility screening in this endangered species. Full article

Currently, the B-cell response patterns induced by viral antigens in avian disease models and their detailed immunological characteristics still require comprehensive elucidation at the single-cell level. In this study, we employed single-cell sequencing (scRNA-seq) and B cell library technology to conduct an in-depth analysis of B cells in the spleens of mice with inactivated duck hepatitis A virus type 1 (DHAV-1) as model antigen. This study aimed to investigate the immunological characteristics of the virus antigen in the mouse model and characteristics of B-Cell Receptors. The results showed that the DHAV-1 group had distinct changes in splenic B cell subset counts, proportions, and intercellular communication. Additionally, an increased trend in communication strength between Gm26917+B and Gm11837+B cells was observed, with enriched expression of C-X-C motif chemokine ligand (CXCL) and lymphotoxin (LT) detected in the DHAV-1 group. Furthermore, the DHAV-1 group exhibited a prominent combination of the IGHV1 family and IGHV3-1/IGHJ3 in the heavy (H) chain variable region. Compared with the CK group (negative control group), the amino acid sequence length and diversity of the CDR3 region in the DHAV-1 group exhibited a decreasing trend. In summary, our findings characterize the immunological features of splenic B cells in mice after immunization with inactivated DHAV-1, and provide a preliminary characterization of DHAV-1-induced B cell transcriptional states and BCR repertoire features, generating testable hypotheses for subsequent mechanistic investigations of B cell-mediated immune responses to viral antigens. Full article

Abnormal intestinal mucosal immunity plays a crucial role in ulcerative colitis (UC). Autotaxin (ATX) can promote T cell migration and was reported to have a regulatory effect on Th17 cells, while sphingosine-1-phosphate (S1P) and its receptors (S1PRs) modulate Th17/Treg balance and inflammation, with S1PR modulators approved for UC. ATX can catalyze sphingosylphosphorylcholine (SPC) to produce S1P; however, the relationship between ATX and S1P/S1PRs in UC is unclear. Understanding the role of ATX-S1P/S1PRs in intestinal immunity can provide new treatment strategies for intestinal inflammatory diseases. Both UC patients and DSS-induced colitic mice showed significantly increased levels of ATX and S1P compared with healthy controls. ATX inhibitor PF8380 treatment led to reduced levels of S1P/S1PRs in colitic mice. Consistent with this, the S1PR antagonist etrasimod was able to alleviate ATX-induced intestinal inflammation, as well as partially restore ATX-induced Th17/Treg imbalance in MLNs and the spleen. In HT-29 and Raw246.7 cells, ATX treatment led to enhanced expression of S1P/S1PRs, with S1PR1 being the most significant. Furthermore, S1PR1 mediates the effect of ATX on Th17/Treg cell differentiation and function in vivo. Therefore, ATX affects the differentiation and function of Th17/Treg cells through S1P/S1PR1 signaling, increased ATX expression leading to Th17/Treg cell imbalance, intestinal mucosal immune dysfunction, and exacerbating intestinal inflammation. Full article

Background: Melanoma is one of the most dangerous types of skin cancer, with its global incidence having surged in recent years. There exists an urgent clinical need for novel therapeutic strategies that combine high efficacy, low toxicity, and multiple mechanisms of action. Methods: This study applies a “Property Optimization and Therapeutic Synergy” strategy, selecting the natural active polysaccharide component, Cordyceps polysaccharides (WCP), as a functional carrier to encapsulate the broad-spectrum chemotherapeutic agent, 10-Hydroxycamptothecin (10HCPT, HCPT). Leveraging non-covalent interactions between the two components, a self-assembly nanoscale drug delivery system (H-W NPs) with high stability and dual antitumor activity was constructed to achieve more efficient and precise antitumor effects. Results: The H-W NPs demonstrated outstanding antitumor efficacy both in vitro and in vivo. The H-W NPs achieved a threefold increase in the inhibition rate against B16-F10 cells compared to free HCPT in vitro and demonstrated a remarkable tumor inhibition rate of 95.08% in vivo. The therapeutic effect may be attributed to the dual antitumor mechanisms of the H-W NPs. Mechanistic studies revealed a synergistic dual-mode of action driving this potent efficacy. Firstly, H-W NPs efficiently induced caspase-3-mediated apoptosis in tumor cells. RNA sequencing analysis suggested the involvement of pathways related to cell cycle arrest and apoptosis. Additionally, H-W NPs promoted the expansion and activation of CD8⁺ T cells in the spleen. These activated cytotoxic T cells reinforced the apoptotic cascade, effectively amplifying the caspase-3-mediated death signal. Conclusions: In summary, the self-assembly nanoscale drug system achieved potent antitumor efficacy through the synergistic action of direct tumor cell killing and immune modulation, offering a highly promising strategy for the development of novel formulations against melanoma. Full article

Immune thrombocytopenia (ITP) is an autoimmune disorder characterized by immune-mediated platelet destruction, resulting in a platelet count below 100 × 10⁹/L and an increased risk of bleeding complications that significantly impair quality of life. Despite advances in ITP management, the unpredictable and heterogeneous nature of the disease continues to challenge treatment selection. This review compares the efficacy, safety, and mechanisms of action of Fostamatinib, Flebogamma DIF, and Romiplostim in adult and pediatric ITP patients. Peer-reviewed studies published over the past 20 years, including randomized and non-randomized clinical trials, observational studies, and real-world evidence, were screened for relevance, with data extracted on dosage, response rates, safety outcomes, and patient characteristics. The sample sizes varied across studies and were reported when available. Study quality and risk of bias were assessed using the ROBINS-I tools. Flebogamma DIF produced rapid increases in platelet count, although its effects were transient. Fostamatinib, the only oral spleen tyrosine kinase inhibitor approved for ITP, was reported to have demonstrated clinical benefit in adults with refractory disease but was contraindicated in pediatric populations. Romiplostim was reported to show sustained platelet responses and facilitate treatment-free remission, particularly in chronic ITP, with an elevated risk of thrombosis. Overall, these therapies offer distinct advantages depending on disease chronicity, patient age, comorbidities, and treatment history. This review underscores the importance of personalized treatment strategies and highlights the need for further long-term, comparative studies to guide evidence-based ITP management. Full article