The metabolomic alterations underlying adolescent myopia progression remain unclear. In this prospective school-based cohort, 186 adolescents (93 progressors and 93 non-progressors) were followed for 12 months and randomly split into a training (n = 150) and an internal validation set (n = 36). Baseline serum samples were profiled using untargeted UHPLC-MS/MS. LASSO, Boruta, and RFE were used to develop a metabolite panel, which was subsequently evaluated in the internal validation set. Hartley guinea pigs (n = 15 per group) were randomized to control, form-deprivation myopia (FDM), and three branched-chain amino acid (BCAA) dose groups (412, 2060, or 4120 mg/kg per day) and received BCAAs by oral gavage for 4 weeks. Refraction, axial length, and scleral thickness were assessed. Human scleral fibroblasts were treated with TGF-β and BCAAs, with mTORC1 or LAT1 inhibition. In the human cohort, we identified 94 endogenous differential metabolites, with BCAA-related metabolism being the most prominently perturbed. A 22-metabolite panel achieved AUROCs of 0.82 and 0.86 in the training and internal validation sets, respectively. L-leucyl-L-alanine was the strongest protective marker (OR = 0.15, 95% CI 0.04‒0.40). BCAAs attenuated axial elongation and preserved scleral thickness in FDM guinea pigs. LAT1 may be involved in BCAA-associated mTORC1 activation, whereas BCAAs may suppress TGF-β/SMAD/FOXO1-associated myofibroblast transdifferentiation. Together, these findings identify a serum metabolomic signature associated with adolescent myopia progression and an internally validated 22-metabolite panel for risk stratification that requires external validation.
Streptococcus suis is an important zoonotic pathogen that threatens global public health and the swine industry. The emergence of multidrug-resistant strains, extensive genetic and antigenic diversity, and limited cross-serotype protection of current vaccines remain major challenges for effective prevention and control. This review summarizes recent advances in the global epidemiology and transmission dynamics of S. suis, integrates molecular mechanisms underlying the sequential progression of infection from initial mucosal colonization to immune evasion and subsequent systemic dissemination, and discusses emerging antibiotic alternative approaches for controlling S. suis infections. Global epidemiological evidence indicates that S. suis infections have been reported in humans and pigs across all continents except Antarctica, with substantial geographic variations in predominant serotypes and sequence types. Mechanistically, S. suis employs diverse virulence factors throughout infection, with dynamic capsular polysaccharide expression contributing to adaptation between colonizing and invasive states, while suilysin promotes epithelial barrier disruption and inflammatory activation, collectively facilitating immune evasion and systemic dissemination. Furthermore, several alternative strategies, including antibiotic adjuvants, anti-virulence agents, phage-derived therapeutics, host-directed therapies, and novel vaccines, have emerged as promising approaches to combat S. suis infections. These insights may facilitate the development of effective interventions against genetically and antigenically diverse S. suis populations while reducing antibiotic reliance and addressing antimicrobial resistance.
Positive-strand RNA viruses remodel host endomembranes to form replication organelles (ROs). However, the membrane sources for coronavirus double-membrane vesicle (DMV) biogenesis remain unclear. Here, we used proteomic, genetic, pharmacological, and imaging approaches to identify the central membrane supply route for DMV formation. APEX2-based proximity proteomics first identified candidate pathways, followed by functional validation using siRNA knockdown, CRISPR/Cas9 editing, and Golgicide A (GCA) inhibition. DMV formation and viral RNA synthesis were assessed by electron microscopy, immunofluorescence, and qRT-PCR. Golgi‒DMV association was analyzed by sucrose gradient fractionation and confocal imaging. In vivo relevance was evaluated in C57BL/6J mice infected with mouse hepatitis virus (MHV) and treated with GCA (intraperitoneal, 50 mg/kg; n = 4–5 per group). We showed that coronaviruses hijack the host GBF1-ARF1-COPI machinery to redirect Golgi-derived membranes to viral replication sites for DMV formation and expansion. Disruption of this pathway markedly impairs viral RNA synthesis, DMV biogenesis, and replication of multiple coronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Pharmacological inhibition of Golgi-specific Brefeldin A-resistance factor 1 (GBF1) suppressed viral replication and pathology in vivo. Together, our findings reveal a previously unrecognized Golgi-derived membrane supply route for coronavirus ROs and highlight the GBF1-ARF1-COPI pathway as a potential host-directed target for broad-spectrum antiviral therapies.
Immunology stands at the forefront of modern biomedical research, driving transformative advances in our understanding of human health and disease. The integration of fundamental scientific discoveries with clinical translation continues to reshape therapeutic strategies for cancer, inflammatory conditions, infectious diseases, and a wide range of immune-mediated disorders. In this dialogue, we speak with Professor Miriam Merad, a global leader in myeloid cell research. Professor Merad reflects on her interdisciplinary career spanning three continents, her landmark discoveries in macrophage biology, the challenges and opportunities in translational immunology, and her vision for the future of global immunological research. This conversation highlights the synergy between diverse research cultures, data-driven scientific discovery, and the pursuit of meaningful clinical impact—core principles that define transformative immunology.
Pseudomonas aeruginosa (PA) is a gram-negative opportunistic bacterium that frequently causes nosocomial infections. Its capacity to evolve from an acute infection characterized by rapid tissue damage to a persistent chronic infection that is difficult to eradicate leads to reduced antibiotic efficacy, disease deterioration, and poor prognosis. This review dissects two major driving forces—intrinsic regulatory networks and adaptive evolution under selective pressures underlying this transition, defines the central role of biofilms, and explores therapeutic implications. Under pressures such as the immune system and antibiotics, PA undergoes lifestyle transition, metabolic reprogramming, and immune evasion through the quorum-sensing (QS), Gac-Rsm, and c-di-GMP networks. Based on these mechanisms, therapeutic approaches targeting the acute-to-chronic infection transition have been developed, including anti-virulence therapy, anti-biofilm strategies, anti-persister strategies, immunomodulatory therapy, and phage therapy. Although these novel therapeutic strategies have demonstrated great potential in preclinical models of chronic persistent infections, their clinical applications remain limited. Future studies should further elucidate the dynamic crosstalk among the core regulatory systems within the host microenvironment and develop intervention strategies targeting key nodes in the acute-to-chronic transition. Such advances hold great promise for eradicating chronic persistent infections and ultimately improving clinical outcomes for patients.
The continued emergence of diverse beta-coronaviruses (β-CoVs) from animal reservoirs underscores the urgent need for broad-spectrum mucosal vaccines, yet their development is hindered by physiological barriers, rapid antigen clearance, and suboptimal multivalent display strategies. To address this, we intranasally immunized BALB/c mice (n = 3 or 4 per group) with 5 or 15 nm gold nanoparticles (AuNPs) displaying the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor-binding domain (RBD, 5 μg per mouse) and adjuvanted with the STING agonist cGAMP (20 μg per mouse). Guided by the serotyping of STING agonist-elicited neutralizing responses, we generated AuNP-Mix co-displaying six representative β-CoV RBDs for comparative immunological evaluation against single-RBD AuNP formulations. Antigen delivery, pulmonary retention, mucosal and systemic immune responses were evaluated via in vivo imaging, enzyme-linked immunosorbent assays, pseudovirus neutralization assays, flow cytometry, immunofluorescence, and challenge studies. We demonstrated that 5 nm AuNP-RBD adjuvanted with cGAMP exhibited superior mucosal penetration and lung retention, induced robust local and systemic immunity, and elicited cross-neutralizing antibodies against SARS-CoV-2 variants, SARS-CoV, and SARS-related coronaviruses. Intranasal immunization with AuNP-Mix elicited broader mucosal/systemic immune responses than those achieved with simple RBD mixtures, as demonstrated by its potent neutralizing activity against SARS-CoV-2 variants, SARS-CoV, Middle East respiratory syndrome coronavirus (MERS-CoV), and divergent MERS-like viruses. These findings establish AuNP-based mosaic display as a versatile platform for next-generation mucosal vaccines.
Targeted protein degradation (TPD) holds significant therapeutic potential over conventional biologics. However, existing platforms are constrained by their reliance on host protein degradation machinery and limited durability, rendering them inadequate for chronic diseases requiring sustained treatment. Here, we constructed a chimeric antigen receptor (CAR) incorporating the TNFR1 ectodomain as the antigen-binding domain (TNFR1T cells) and used CRISPR-mediated knockout of BCOR and ZC3H12A to generate persistent TNFR1TIF cells. TNF binding, endocytosis, and degradation were assessed in vitro by flow cytometry and immunofluorescence. Immunocompetent syngeneic, Tnf−/−, and hTNF-tg RA mice (n = 4–9) were used in vivo to evaluate engraftment, persistence, and disease severity (clinical scoring and grip strength), with adalimumab (Humira, 1 or 10 mg/kg) as a comparator. Safety was assessed by intravenous Listeria monocytogenes challenge (1 × 104 colony-forming units [CFU], n = 6) and Thy1.1-based depletion (0.25 mg, n = 3). We found that TNFR1T cells specifically bound, endocytosed, and degraded soluble TNF in vitro. Without lymphodepletion preconditioning, TNFR1TIF cells expanded and persisted for one year in immunocompetent mice. A single infusion into hTNF-tg mice reduced serum hTNF to near wild-type levels, preventing and treating all stages of RA with superior efficacy and durability than the repeated high-dose adalimumab. Antibacterial defense remained uncompromised, and anti-Thy1.1 antibody efficiently eliminated TNFR1TIF cells in vivo. This approach extends CAR-T cell targeting from cellular antigens to soluble extracellular proteins, establishing a host-machinery-independent, durable cellular-TPD platform for chronic inflammatory diseases.
Klebsiella pneumoniae is an important opportunistic pathogen, and its capsular polysaccharides are key virulence factors. Capsular typing is important for epidemiological surveillance, clinical diagnosis, and therapy development; however, methods for capsule-targeted detection remain limited. Here, we engineered a modular bioluminescent reporter phage platform leveraging bacteriophage receptor-binding proteins for specific K. pneumoniae capsular typing. Initially, the K2-specific bacteriophage ΦRCIP0109 was engineered with the nluc reporter gene to generate ΦRCIP0109::nluc for K2 detection. Using ΦRCIP0109::nluc as the chassis, we engineered reporter phages by swapping receptor-binding proteins (RBPs), thereby expanding the detection range to four additional clinically important K types (K64, K47, K1, and K57). Reporter phage specificity and sensitivity were evaluated against clinical isolates and in simulated polymicrobial environments. Directed evolution was employed to introduce RBP mutations for enhanced phage adsorption. These five reporter phages achieved 100% specificity against clinical isolates, detecting concentrations as low as 10 CFU/mL within 3.5–5.5 h and successfully differentiating host strains in polymicrobial synthetic urine. Directed evolution of an RBP yielded a 10- to 100-fold increase in luminescence compared to that of the wild-type phage. Together, these advances establish a scalable platform that can be expanded to additional capsular types and the detection of other pathogens, representing a platform with clear potential for integration into point-of-care diagnostics, guiding targeted antimicrobial therapy and precision phage therapy.
Tracking mature neutrophils remains challenging due to the lack of reliable cell surface markers. Although CD101 is a promising candidate as a marker for mature neutrophils, its stability under pathological conditions is unclear. Here, we developed a novel CD101-tdTomato reporter mouse model to specifically identify mature neutrophils. CD101 expression, tdTomato fluorescence, and neutrophil physiology were characterized by flow cytometry across peripheral tissues under homeostasis and by a transwell migration assay, in parallel with morphological features assessed by Wright-Giemsa staining. To assess marker stability, CD101-tdTomato and wild-type (WT) mice were treated with lipopolysaccharide (LPS) (200 ng, intraperitoneally, n = 8, 2 h), influenza virus (5 × 103 PFU/mL, intratracheally, days 1 and 5, n = 6), or granulocyte colony-stimulating factor (G-CSF) (2.5 μg, intravenously, 2–6 h, n = 5), followed by flow cytometry analyses. CD101-tdTomato mice were crossed with Lysozyme-GFP mice to validate reporter specificity across myeloid subsets. We confirmed that CD101+ and tdTomato+ neutrophils display identical characteristics of mature neutrophils, including poly-segmented nuclei, cell size, and key functional properties under homeostasis. Under pathological challenges, the decrease in CD101 expression was not attributed to shedding or degradation of the protein, supporting its robustness under inflammatory conditions. Using CD101-tdTomato × Lysozyme-GFP mice, we established a dual-reporter system that enables precise distinction of mature neutrophils from total myeloid cells. Collectively, our findings support CD101 as a robust and reliable marker of neutrophil maturity, providing a foundation for its future applications in spatial transcriptomics and lineage tracing studies of the heterogeneity and functions of neutrophils.
Stem cell-derived organoids are emerging as living therapeutics for repairing or replacing damaged tissues and organs, offering new opportunities for regenerative medicine. However, their clinical translation remains limited by immune rejection, insufficient functional integration, and low manufacturing throughput. In this review, we summarize recent advances in engineering organoids toward implantable living therapeutics by examining how hydrogel engineering, microfluidic chips, three-dimensional (3D) bioprinting, and synthetic biology overcome key translational barriers and facilitate the in vivo application of organoids. We further discuss the existing regulatory frameworks supporting the clinical translation of organoid-based therapies. Current evidence indicates that these bioengineering strategies enhance graft survival, tissue integration, and therapeutic efficacy through complementary mechanisms including immunomodulatory signals, biomimetic microenvironments, hierarchical tissue architectures, and programmable cellular functions. Importantly, engineered organoids have been successfully applied in restoring hair-bearing skin, repairing colonic epithelial continuity, bridging critical-size bone defects, and correcting metabolic disorders. Despite these advances, significant challenges remain in quality-control standardization, scalable manufacturing, regulatory harmonization, and long-term safety assessment. Future studies integrating artificial intelligence (AI) and intelligent living devices are warranted to facilitate high-fidelity manufacturing, quality inspection, real-time monitoring, and closed-loop therapeutic regulation in clinical practice. Collectively, engineered organoids represent a promising frontier for new-generation transplantation and regenerative medicine.
Studies on dengue virus (DENV) infection during pregnancy show that symptomatic dengue is associated with adverse fetal outcomes, but the mechanisms remain unclear. Our previous research indicated that neutrophils contribute to intrauterine growth restriction (IUGR) by damaging placental vasculature, yet the molecular mechanisms underlying this microvascular injury remain poorly defined. To address this, this study examined placental microvascular ferroptosis by bulk and single-cell transcriptomic reanalyses, immunofluorescence staining, and Western blotting in DENV-2-infected E18.5 placentas from pregnant Ifnar1u2212/u2212 C57BL/6J mice (n = 5 per group). Causality was investigated by assessing fetal weight and microvascular damage assessment following administration of ferroptosis inhibitor, neutrophil extracellular trap formation (NETosis) inhibitor, or glutathione supplementation. We observed that ferroptosis in placental vascular endothelial cells was accompanied by vascular disruption and NETosis in our IUGR mouse model. Pharmacological inhibition of ferroptosis alleviated placental injury and restored fetal weight, directly linking ferroptosis to DENV-2-induced IUGR. Moreover, increased 4-hydroxynonenal (4-HNE) levels and decreased glutathione peroxidase 4 (GPX4) levels were detected in the placenta, indicating oxidative stressu2013driven lipid peroxidation. Glutathione supplementation attenuated ferroptosis and IUGR. Finally, NETosis inhibition reduced placental lipid peroxidation and vascular injury, suggesting that excessive NETosis initially triggers oxidative stress. Taken together, our data suggest that ferroptosis in vascular endothelial cells induced by neutrophil NETosis is the driving cause of IUGR in DENV infections. This study reveals the mechanism underlying DENV-associated adverse pregnancy outcomes and provides potential therapeutic insights.
Regulatory T cell (Treg) instability has evolved from a debated artifact into a central concept in immunology, though the mechanisms driving the transition from stable suppressor to pathogenic ex-regulatory T cell (ex-Treg) remain unclear. This review synthesizes findings from fate-mapping, epigenetic, and metabolic studies in mouse models and human systems, tracing the ex-Treg concept through three phases: controversy, epigenetic reconciliation, and current views. The Treg-specific demethylated region (TSDR) distinguishes truly committed Tregs from transient forkhead box P3 (Foxp3) expressers. Three interdependent factors—environmental stress (interleukin-1β [IL-1β], interleukin-6 [IL-6]), epigenetic erasure (ten-eleven translocation 2 [TET2] dysfunction leading to TSDR hypermethylation), and metabolic rewiring (mechanistic target of rapamycin [mTOR]-driven glycolysis)—cooperatively drive ex-Treg conversion. The “Self vs Non-self” hypothesis frames instability as an evolved checkpoint balancing tolerance with host defense. Ex-Tregs contribute to autoimmunity and aging but also offer therapeutic opportunities in cancer. Key uncertainties persist regarding ex-Treg heterogeneity, epigenetic reversibility, and translation to humans. Future directions should focus on biomarker discovery and engineering epigenetically stable Tregs for therapy. Collectively, these advances position Treg instability as a pivotal regulator of immune homeostasis and highlight the promise of therapeutically targeting this pathway to advance treatments for autoimmunity, aging, and cancer.
