Search Results (76)

Background: Aluminum-adjuvanted hepatitis B vaccines effectively prevent primary HBV infection but predominantly favor humoral immunity and have limited capacity to induce strong cellular responses. To improve immune-response quality, we developed a Ganoderma lucidum polysaccharide-modified hybrid nanoparticle adjuvant (GHNP) for hepatitis B surface antigen (HBsAg) vaccination. Methods: Cationic PEG-PCL/DOTAP hybrid nanoparticles (HNPs) were surface-modified with Ganoderma lucidum polysaccharide (GLP) at different mass ratios and characterized by particle size, polydispersity index, zeta potential, and transmission electron microscopy. Antigen uptake, RAW264.7 cell viability, MHC II expression, and TLR2/4-associated TNF-α secretion were evaluated in vitro. C57BL/6 mice were immunized intramuscularly with PBS, HBsAg, HBsAg + Aluminum, HBsAg@HNP, or HBsAg@GHNP6 on Days 0, 14, and 28. HBsAg-specific antibodies, splenocyte activation, proliferation, and cytokine secretion were assessed. Results: GHNP6 showed suitable physicochemical properties, acceptable cytocompatibility at concentrations up to 10 μg·mL⁻¹, and efficient model-antigen uptake. GHNP increased MHC II expression in RAW264.7 cells, and TLR2/4 inhibition reduced nanoparticle-associated TNF-α secretion, supporting involvement of these pathways in immune activation. In vivo, HBsAg@GHNP6 induced robust HBsAg-specific IgG responses and a higher IgG2a/IgG1 ratio than HBsAg@HNP and HBsAg + Aluminum. It also produced the highest splenocyte proliferation after HBsAg restimulation, with lower IL-6 secretion than HBsAg + Aluminum and moderately increased IFN-γ compared with PBS and free HBsAg. Conclusions: GHNP integrates nanoparticle-assisted antigen delivery with GLP-mediated immunomodulation and represents a promising strategy for enhancing cellular immune responses while maintaining humoral immunity in HBsAg vaccination. Full article

Background: Early-life microbial exposures shape immune maturation and can influence later susceptibility to allergic airway disease. While probiotics can modulate host immunity, determining whether maternal probiotic supplementation alone promotes changes in offspring microbiomes and the immune functions that reduce allergic disease remains incompletely defined. Methods: Female mice were gavaged with Lactobacillus johnsonii (Lj) daily for 7 days prior to mating, and then twice weekly until delivery. Offspring were sensitized and challenged with cockroach allergen (CRA), beginning at 5 weeks of age, for 3 weeks to initiate asthmatic-type responses. Airway physiology (methacholine-induced airway hyperreactivity; AHR), lung-mucus-related gene expression (Muc5ac, Gob5), and lymph node restimulation responses were assessed. Results: Offspring from Lj-supplemented dams exhibited reduced AHR and decreased induction of Muc5ac and Gob5 following the final CRA challenge, accompanied by diminished Th cell cytokine production upon lymph node restimulation. Maternal Lj supplementation produced a significant shift in offspring cecal microbiome structure at 5 weeks, including altered phylum-level composition and a discrete set of significantly changed OTUs, including Akkermansia. PICRUSt2 analyses predicted coordinated differences in microbial metabolic potential across multiple pathways, consistent with functional reprogramming of the gut microbiome. Bone marrow dendritic cells (BMDC) derived from adult offspring of Lj-supplemented dams showed attenuated inflammatory programming after RSV or TLR7 stimulation, with reduced expression of innate immune cytokines compared to PBS-supplemented dams. Conclusions: The relatively long-term effects were associated with reduced allergic airway disease severity, supporting maternal probiotic supplementation as a potential strategy to lower offsprings’ risk of allergic airway pathology. Full article

Neutrophils are key effectors of innate immunity. Although traditionally regarded as short-lived effector cells, recent evidence suggests they can undergo functional reprogramming after microbial stimulation, leading to trained immunity. We investigated whether lipopolysaccharide (LPS) or Candida albicans β-glucan induces memory-like inflammatory responses in adult peripheral and neonatal cord blood neutrophils. We used complementary molecular and functional approaches to characterize neutrophil responses following in vitro priming with LPS or β-glucan and subsequent LPS restimulation. Cytokine secretion, reactive oxygen species (ROS) production, glycolytic activity (via lactate production and glycolytic enzyme expression), and ERK1/2 and NF-κB signaling pathway activation were assessed. Priming with LPS or β-glucan enhanced neutrophil responsiveness to secondary stimulation, resulting in increased secretion of IL-1β, IL-6, IL-8, CXCL1, and CCL2, together with elevated ROS production. These effects were consistently more pronounced in neonatal than in adult neutrophils. Memory-like neutrophils also exhibited metabolic reprogramming, associated by increased hexokinase-2 and phosphofructokinase-1 mRNA expression, elevated lactate production, and enhanced ERK1/2 and NF-κB activation. Human neutrophils, particularly neonatal cells, develop memory-like characteristics following LPS or β-glucan priming, highlighting their functional and metabolic plasticity and suggesting a role for neutrophil training in early-life immune adaptation. Full article

Reservoir stimulation is essential in enhanced geothermal systems and unconventional reservoirs where low permeability, inadequate fracture connectivity, or near-wellbore damage restricts commercial injection or production. This review evaluates hydraulic fracturing, thermal stimulation, plasma-pulse stimulation, and selected dynamic stimulation technologies. It compares their physical mechanisms, fracture-network development, reservoir applications, permeability enhancement, operational maturity, deployment challenges, and future perspectives. Hydraulic fracturing remains the most mature method for reservoir-scale fracture creation, fracture conductivity, and reservoir connectivity. In enhanced geothermal systems, however, performance depends on the heat-exchange area, distributed flow, thermal sweep, long-term energy recovery, and induced-seismicity control rather than permeability enhancement alone. Thermal stimulation is integral to geothermal reservoir development. Cold-fluid injection generates thermoelastic stress redistribution, enlarges the fracture aperture, activates natural fractures, promotes thermally assisted fracture propagation, and influences thermal breakthrough. Plasma-pulse stimulation, also termed pulsed-power plasma, electrohydraulic, or shock-wave stimulation, provides a low-water method for near-wellbore permeability enhancement, damage bypass, fracture reactivation, and restimulation. Its broader deployment remains constrained by the limited treatment radius, scale-up uncertainty, energy-transfer efficiency, tool durability, completion integrity, and insufficient field validation. Liquid CO₂ phase-transition, propellant, and explosive stimulation provide additional dynamic-loading options with distinct fracture responses, controllability, safety, and technology readiness. Stimulation technologies should therefore be selected according to the dominant reservoir limitation and evaluated using sustained injectivity or productivity, effective reservoir contact, distributed flow, delayed thermal breakthrough, treatment durability, wellbore integrity, and a controlled geomechanical response. Future progress requires hybrid stimulation, coupled thermal–hydraulic–mechanical–chemical (THMC) modeling, integrated monitoring, adaptive control, physics-informed artificial intelligence, digital twins, standardized field validation, and techno-economic and life-cycle assessments. Full article

Background: Chagas disease, caused by Trypanosoma cruzi, remains a major public health problem in Latin America and an emerging concern worldwide. Current chemotherapies show limited efficacy during chronic infection, and no licensed vaccine is currently available. We previously developed the chimeric antigen N-Tc52/TSKb20 as a vaccine candidate against T. cruzi infection. In a murine model, this vaccine induced robust antigen-specific immune response associated with protection shortly after vaccination. Objectives: Here, we investigated the long-term persistence and effector functions of the immune responses elicited by this vaccine candidate. Methods: Both female and male C57BL/6 mice were immunized with three doses of N-Tc52/TSKb20 formulated with QuilA adjuvant. Serum samples collected 170 days post-immunization were analyzed for antigen-specific antibodies by ELISA and for trypanolytic activity against cell-derived trypomastigotes using an in vitro functional assay. Cellular immune responses were evaluated by measuring cytokine production, T cell activation, and memory T cell responses following in vitro re-stimulation with the vaccine antigen or T. cruzi antigens. Results: N-Tc52/TSKb20 vaccination induced a sustained antigen-specific humoral response, characterized by long-lasting IgG2c antibodies and functional activity persisting for up to 170 days post-immunization. In parallel, vaccination promoted long-term activation of antigen-specific CD8⁺ T cells and production of TNF-α and IFN-γ upon antigen re-encounter. A sex-dependent tendency was observed for IL-10, with increased production in vaccinated female mice. Moreover, vaccinated animals exhibited increased frequencies of central and effector memory CD4⁺ and CD8⁺ T cells in response to T. cruzi antigens, with a predominant contribution of CD8⁺ T cells, indicating the establishment of parasite-specific T cell memory. Conclusions: Together, these findings demonstrate that vaccination with N-Tc52/TSKb20 induces a long-lasting Th1-biased immune response characterized by trypanolytic antibodies, functional and durable T cell responses, and parasite-specific memory T cells. This immunological profile supports the potential of N-Tc52/TSKb20 as a promising vaccine candidate for Chagas disease and highlights its capacity to elicit immune mechanisms that have been associated with protection against T. cruzi infection. Full article

In addition to inducing pathogen-specific adaptive immune responses, vaccines can train the innate immune system, thereby providing broader host protection. This concept of trained immunity (TRIM) is well-established in benchtop laboratory science. This review aims to evaluate the current evidence of vaccine-induced TRIM and translate these findings into a clinical context. Various laboratory methods are used to assess TRIM; however, inconsistent results have been reported across non-BCG vaccine studies. Existing analyses lack exploration of the mechanistic basis of vaccine-induced TRIM, particularly epigenetic reprogramming and metabolic rewiring. Patterns emerge between vaccines: live-attenuated vaccines generally induce TRIM, as evidenced by increased inflammatory cytokine production upon restimulation, whereas non-live vaccines tend to demonstrate reduced trained immunity. Such findings are not consistently observed for mRNA vaccines, which show heterogeneous patterns. The limited variety of studies on non-BCG vaccines impacts the reliability of findings. A more comprehensive understanding of the mechanisms and outputs of TRIM induced by specific vaccines could better inform rational vaccine design. Furthermore, various modifiers can alter vaccine-induced TRIM responses, including sequence and route of administration, sex, and age. Consideration of these modifiers has important clinical implications in optimising vaccine administration for enhanced immune protection. Full article

Background: Quiescent cancer cells (QCCs) evade conventional therapies and contribute to minimal residual disease (MRD) and relapse, yet the signaling pathways governing their survival remain poorly understood. Methods: Here, we performed integrative proteomic and phosphoproteomic profiling of triple-negative breast cancer (TNBC) cells transitioning between proliferation and serum removal-induced quiescence, followed by re-stimulation. Results: We identified dynamic remodeling of both proteome and phosphoproteome, with quiescent cells showing downregulation of mitotic drivers and upregulation of extracellular matrix components. Notably, phosphorylation of CK2 substrates was increased during quiescence, and CK2 inhibition using CX-4945 impaired cell survival under nutrient and genotoxic stress, disrupted autophagy, microtubule dynamics, and protein synthesis. Phospho-enrichment and functional assays identified death-associated protein kinase 3 (DAPK3) as a CK2-regulated effector mediating stress-induced apoptosis. In silico analysis confirms a link between high CK2 expression and poor chemotherapy response in basal breast cancer. Conclusions: These findings establish CK2 as a critical survival kinase in QCCs and a potential therapeutic target for MRD eradication in breast cancer. Full article

Background/Objectives: Sporotrichosis is an emerging zoonotic subcutaneous fungal infection with limited therapeutic options, highlighting the need for improved immunomodulatory strategies. CTLA-4 is an inhibitory immune checkpoint that negatively regulates T-cell activation. In this study, we evaluated whether a CTLA-4 antisense oligonucleotide (CTLA-4 ASO) is associated with enhanced immune responses to an adjuvanted recombinant Sporothrix sp. enolase antigen (rSsEno) formulation. Methods: CTLA-4 ASO uptake, cytotoxicity, and gene-silencing activity were assessed in murine splenocytes in vitro. BALB/c mice were immunized with rSsEno formulated with Montanide Gel 01, either alone or in combination with 5 µg CTLA-4 ASO. Antigen-specific serum antibody responses were quantified by ELISA. Splenocytes from immunized mice were restimulated with enolase, and cytokine production (IFN-γ, IL-2, IL-17, and TNF-α) was measured using Cytometric Bead Array (CBA). Results: CTLA-4 ASO was efficiently internalized by splenocytes and was associated with reduced expression of CTLA-4 without detectable cytotoxicity in vitro. Mice receiving the ASO-supplemented formulation developed significantly higher anti-enolase antibody titers compared to those immunized with adjuvant alone. Upon antigen restimulation, splenocytes from ASO-treated mice produced higher levels of IFN-γ, IL-2, TNF-α, and IL-17, consistent with an enhanced recall response characterized by a mixed Th1/Th17 cytokine profile. Conclusions: CTLA-4 ASO was associated with an enhanced recall response characterized by a mixed Th1/Th17 cytokine profile. These findings suggest a potential immunomodulatory effect of CTLA-4 targeting. Further studies incorporating dose optimization, infection challenge models, and appropriate sequence controls are required to determine the specificity and relevance of these effects for protective immunity against sporotrichosis. Full article

Background: Infections caused by multidrug-resistant bacteria and inadequate vaccine coverage against opportunistic pathogens highlight the need for interventions that broadly and durably enhance host defense beyond antigen-specific adaptive immunity. Trained immunity, driven by metabolic and epigenetic reprogramming of innate immune cells, has been predominantly characterized using Bacille Calmette–Guérin and β-glucan, whereas its induction by Gram-negative bacteria remains poorly defined. To address this gap, we aimed to determine whether heat-killed Klebsiella pneumoniae (HK Kp) induces trained immunity through metabolic and hematopoietic reprogramming to confer heterologous antibacterial protection. Methods: HK Kp-trained murine bone marrow-derived macrophages and HK Kp-immunized C57BL/6 mice were employed to interrogate functional, metabolic, and transcriptomic reprogramming in vitro, hematopoietic progenitor remodeling in vivo, and protective efficacy against systemic Salmonella Typhimurium and Staphylococcus aureus infection. Results: HK Kp-trained macrophages showed markedly enhanced IL-1β secretion across all restimulation conditions, stimulus-dependent amplification of TNF-α responses, increased phagocytosis, and improved intracellular control of S. typhimurium, together with sustained upregulation of the glycolytic enzymes-encoding genes Hk2 and Pfkfb3. Transcriptomic profiling revealed extensive reprogramming enriched in glycolysis/gluconeogenesis and hematopoietic cell lineage pathways. In vivo, HK Kp immunization shifted bone marrow stem/progenitor compartments toward a myeloid-biased state. HK Kp-trained mice challenged with lethal S. typhimurium or S. aureus exhibited less weight loss, improved survival rates, and reduced bacterial burdens. Conclusions: Inactivated K. pneumoniae orchestrates metabolic and hematopoietic reprogramming to establish enhanced innate immune responsiveness and confer heterologous protection in murine S. typhimurium and S. aureus sepsis models, supporting its potential as a potent inducer of trained immunity. These findings establish HK Kp-based trained immunity as a promising strategy for combating multidrug-resistant and vaccine-evading pathogens. Full article

Background/Objectives: The gut microbiome has an important role in immune regulation, and dietary interventions that support a balanced microbiota may help to prevent the development of allergic asthma. Dietary fibers can beneficially affect the intestinal microbiome, but due to the diversity of fiber types, the effects differ. In this study, we investigate the preventive effects of two mixes of short-chain and long-chain (1:1 and 9:1 ratio) fructo-oligosaccharides (FOS) in a mouse model of house dust mite (HDM)-induced allergic asthma. Methods: BALB/c mice received FOS-supplemented (1% w/w) diets before and during intranasal exposures to HDM. Endpoint airway hyperreactivity measurements were performed, followed by the collection of bronchoalveolar lavage fluid (BALF), lung, serum and cecum content. Fecal microbiome composition was determined by DNA sequencing and short-chain fatty acid (SCFA) levels were determined in the cecum, serum and lung. Results: Fecal microbiome analyses revealed an increased abundance of Prevotellaceae after FOS1:1 supplementation in HDM-allergic mice. Additionally, FOS1:1 protected against an HDM-induced increase in basal airway resistance. Both FOS1:1 and FOS9:1 restored the systemic acetate levels in HDM-allergic mice. The two FOS supplementations did not affect HDM-induced inflammatory cell influx in the BALF. However, FOS1:1 increased the frequency of Th1-cells and prevented an HDM-induced increase in the Th2/Th1 balance. Upon ex vivo restimulation with HDM, lung cell suspensions of FOS1:1-fed mice produced less type 2-related cytokines compared to control-supplemented mice, and FOS9:1 followed a similar pattern. Conclusions: Specific short-chain and long-chain FOS ratios differentially affect the microbiome and immune system in a mouse model with HDM-induced allergic airway inflammation. Dietary supplementation with FOS1:1 shifts the immune response away from type 2, suggesting that dietary fibers like FOS1:1 may contribute as a part of a broader strategy to modulate HDM-induced allergic asthma. Full article

Background/Objectives: Porcine reproductive and respiratory syndrome virus (PRRSV) remains one of the most economically significant pathogens in the global swine industry. Despite the availability of commercial vaccines for over three decades, they fail to induce sterile immunity and often provide inconsistent protection against heterologous PRRSV strains. This study aimed to predict vaccine immunogenicity by detecting strain-specific immune responses that related to an immune correlate of protection (CoP) against different PRRSV-2 strains. Methods: Post-weaning pigs were vaccinated with five commercially available PRRSV-2 vaccines or received sterile PBS injection as a control. At 28 days post-vaccination (dpv), all pigs were humanely euthanized for large-volume blood collection to isolate peripheral blood mononuclear cells (PBMCs) and plasma, establishing the immune bank. PBMCs and plasma from each group were then tested against six PRRSV-2 strains to evaluate immune responses. In addition, T cell epitope coverage between vaccine and field PRRSV-2 strains was assessed using the EpiCC (in silico) tool to enhance predictive capacity. Results: While neutralizing antibodies were undetectable in all vaccinated pigs at 28 dpv, PRRSV-specific IFNγ–producing cells were detected at various levels in each vaccinated group following restimulation with different PRRSV-2 strains. Additionally, a positive correlation was observed for the EpiCC coverage of the N gene and mean IFNγ responses to VR2332 (SLA class I and II) and NC24-6 (SLA class II). Conclusions: The PRRSV immune bank demonstrated potential as a tool for predicting vaccine immunogenicity against different PRRSV-2 strains and EpiCC provides additional information on T cell epitope cross conservation. The combined approach may provide a valuable framework for selecting PRRSV vaccines for more effective prevention and control in endemic areas. Full article

Background/Objectives: Immunotherapy has shown promising results in some cancers, but its efficacy remains limited in pancreatic ductal adenocarcinoma (PDAC). Vaccines in nanoparticle form (nanovaccines) can incorporate immunostimulating components to induce a potent immune response. As mesothelin (MSLN) is a tumor-associated antigen overexpressed in PDAC, we evaluated the effect of MSLN nanovaccine in a syngeneic orthotopic KPC-PDAC mouse model. Methods: An MSLN peptide combining three MSLN epitopes and two adjuvants, poly I:C and R848, was encapsulated in PLGA–chitosan nanoparticles to generate the nanovaccine. Results: The MSLN nanovaccine was successfully taken up by dendritic cells in vitro and was found in inguinal lymph nodes 24 h after subcutaneous injection into C57BL/6 mice. Nanovaccine re-stimulation of splenocytes from vaccinated mice led to increased levels of interferon-γ in vitro compared to unstimulated splenocytes. Higher levels of MSLN-specific IgM and IgG antibodies were detected in the serum of vaccinated mice compared to that of control mice. Three vaccination regimens were tested: a prophylactic scheme that included vaccination before tumor induction and two therapeutic schemes involving early and late vaccination after tumor cell inoculation. MSLN nanovaccination inhibited KPC tumor progression and metastasis and induced higher CD8⁺ T cell infiltration in the tumor that developed in response to prophylactic and early therapeutic schedules but not in response to a later vaccination approach. Although the nanovaccine treatment elicited higher humoral and cellular antigen-specific responses in tumor-bearing mice for both vaccination strategies, the therapeutic vaccination also increased the expression of exhaustion markers in CD8⁺ T cells. Conclusions: Our results support the relevance of an MSLN-based nanovaccine as a new immunotherapy treatment for PDAC and propose an innovative method of vaccine delivery using NPs. Full article

Low-energy ion bombardment (IB) has emerged as a promising, maskless nanofabrication tool for quasi-periodic nanoripples, marked by a high throughput and low cost. As templates, these IB-induced, self-organized surface nanoripples have shown potential for applications in diverse fields. However, the challenge of tailoring the ordering of these ripple patterns is preventing the widespread application of IB. Moreover, the enhancement of the ordering of these self-organized nanostructures involves the fundamental academic questions of nanoripple coupling (or superimposition) and guided self-organization. This review first focuses on the experimental progress made in developing representative strategies for the ordering enhancement of IB-induced nanoripples in terms of ion beams and targets. Second, we present our understanding of these developments from the perspectives of ripple superposition and guided self-organization. In particular, the basic conditions for ripple superposition under the non-conservation of mass are deduced based on the common features of the results from rocking bombardments of a single material and the bombardment of bilayer systems, providing insight into the mechanisms at play and deepening our understanding of these experimental observations. Finally, areas for future research are given, with the aim of improving ripple ordering from the viewpoints of ripple superimposition and guided self-organization. All this may re-stimulate interest in this field and will be of importance in advancing the academic research and practical applications of IB-induced nanopatterns. Full article

We investigated whether microalgae or linseed supply during the early postpartum period affects ovarian restimulation and supports the first postpartum ovulation in lactating anovulatory goats. Thirty-eight An-glo-Nubian-crossbred adult goats were allocated into three groups, one with a control diet (n = 12), fed a total mixed ration (TMR) comprising chopped elephant grass and concentrate; an algal diet (n = 13), fed TMR + green microalgae (1% dry matter); and a linseed diet (n = 13), TMR + linseed (12% dry matter). Supplements were furnished from the second to fifth week (time of weaning). Goats were estrus synchronized on day 40 by insertion of an intravaginal CIDR device for 5 days, after which 0.075 mg PGF2α was applied to induce ovulation, and estrus was monitored for 72 h. From the 5th–15th day of ovulation induction, the corpus luteum (CL) area and progesterone rate were monitored. The algal and linseed groups showed lower feed intake (p < 0.001) and higher (p < 0.001) triglyceride levels/follicle numbers, respectively. After estrus induction, no differences were observed in estrus response; however, the linseed group showed more and larger growing follicles (p = 0.016 and p < 0.01), a higher ovulation rate (p < 0.05), a larger CL area (p < 0.05), and higher progesterone levels (p < 0.001). Linseed after delivery stimulates follicular growth before and after ovulation induction, favoring better CL quality during the first ovulation. Full article

CD4+ T cells are considered the main orchestrators of autoimmune diseases. Their disruptive effect on CD4+ T cell differentiation and the imbalance between T helper cell populations can be most accurately determined using experimental autoimmune encephalomyelitis (EAE) as an animal model of multiple sclerosis (MS). One epigenetic factor known to promote autoimmune inflammation is miRNA-155 (miR-155), which is significantly upregulated in inflammatory T cells. The aim of the present study was to profile the transcriptome of immunized mice and determine their gene expression levels based on mRNA and miRNA sequencing. No statistically significant differences in miRNA profile were observed; however, substantial changes in gene expression between miRNA-155 knockout (KO) mice and WT were noted. In miR-155 KO mice, mRNA expression in CD4+ T cells changed in response to immunization with the myeloid antigen MOG_35-55. After restimulation with MOG_35-55, increased Ffar1 (free fatty acid receptor 1) and Scg2 (secretogranin-2) expression were noted in the CD4+ T cells of miR-155-deficient mice; this is an example of an alternative response to antigen stimulation. Full article