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Viral (LCMV) coinfection decreases the number of pathogenic Th17 cells in the CNS and prevents demyelinating disease 2309860
Abstract Introduction Multiple sclerosis is a chronic autoimmune demyelinating disease impacting 2.9 million people worldwide. Despite the significant advances in understanding the pathogenicity of CD4+ T cells, there is no cure. Infections have often been considered a trigger for the development of autoimmune diseases, yet infections can also have potential protective roles. Here, we investigate the impact of viral co-infection on demyelinating disease. Methods We found that infecting C57Bl/6 mice with lymphocytic choriomeningitis virus (LCMV) during experimental autoimmune encephalomyelitis (EAE) leads to complete protection from demyelinating disease onset and progressive paralysis compared to uninfected controls. Results Overall, there was a substantial decrease in the numbers of pathogenic Th17 CD4 T cells. Using 2D-micropipette adhesion frequency assay, we demonstrated reduction in the overall number of myelin specific T cells and their affinity for MOG35-55. In adoptive transfer experiments, LCMV infection also prevented demyelinating disease induced by 2D2 TCR Tg CD4+ T cells. While viral coinfection prevented disease, Type I interferon was not the causative mechanism. Conclusion Our work indicates that viral co-infection can decrease the number and affinity of pathogenic, myelin specific CD4 T cells in the CNS. Funding Source NA Topic Categories Neuroimmunology (NEUR)
Neutrophil Extracellular Traps Prime B Cell—Mediated Fibrogenesis in MASH by Silencing HES1 and Promoting Plasma Cell Differentiation 2261119
Abstract Introduction Metabolic dysfunction—associated steatohepatitis (MASH) is a severe form of metabolic dysfunction—associated steatotic liver disease (MASLD) that can progress to fibrosis, cirrhosis, and hepatocellular carcinoma. Although the FDA-approved Resmetirom shows promise in reversing MASLD, its efficacy in advanced fibrosis remains limited. MASH-fibrosis is difficult to treat partly due to immune microenvironment alterations that sustain inflammation and scarring. B cells have emerged as critical contributors to MASH-fibrosis, yet their mechanisms remain incompletely understood. Here, we investigated how neutrophil extracellular traps (NETs) modulate B-cell function to promote MASH-associated fibrosis. Methods C57BL/6 mice were fed a Western diet (WD) for 40 weeks to induce MASH-fibrosis. NETs were inhibited using neutrophil-specific Pad4 knockout (Pad4fl/fl LysM-Cre) or daily DNase I treatment. Fibrosis was assessed by Sirius Red and Masson staining; cytokines and chemokines were measured by Luminex assays. Hepatic leukocytes were analyzed by scRNA-seq and flow cytometry. Plasma B cells and HES1 expression were evaluated by flow cytometry and in vitro NET stimulation assays. Mature B cells were depleted in vivo using muMT mice. Results WD-induced MASH-fibrosis increased hepatic CD19+CD138+ plasma B cells and suppressed HES1 expression, both reversed by NET inhibition. In vitro, NETs directly suppressed HES1 and promoted plasma-cell differentiation, an effect abolished by DNase I. scRNA-seq revealed broad immune changes across nine hepatic immune cell types, with B cells showing a prominent transcriptomic shift that was normalized by NET inhibition. Both NET blockade and B-cell depletion reduced hepatic fibrosis, inflammation, and cytokine levels. Conclusion NETs promote B-cell—mediated fibrogenesis in MASH by silencing HES1 and driving plasma-cell differentiation. Targeting NET-induced B-cell reprogramming offers a promising therapeutic strategy to limit liver inflammation and fibrosis in MASH. Funding Source n/a Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)
Introduction of a remediation process to support student learning in veterinary immunology 2308929
Abstract Introduction The veterinary curriculum is challenging, and individual students sometimes need additional opportunities to assimilate content. Accordingly, we developed and implemented a structured remediation process, called Upgrade, for veterinary immunology to support student learning by promoting content mastery, encouraging self-reflective learning strategies, and mitigating academic stress. This formal remediation process occurs during the semester and provides a structure for students to further engage with the content and have a second chance to demonstrate mastery. Methods Students are eligible any time they score below 70% on an exam; there are three exams, and students must achieve an overall grade ≥ 70% to pass the course. The remediation process has two levels depending on the student’s exam score: 1) students scoring 60—69.99% complete annotations of incorrect exam questions, and 2) students scoring < 60% complete exam annotations, meet individually with the course coordinator, and take an online exam on content specific to the unit. Students who successfully complete the process have their exam score amended to a 70% (or the higher of the two exam scores, if they take the online exam but score < 70%). This process was implemented in Spring 2025, and students were surveyed at the end of the academic year. Results Students who remediated an exam found the process to be helpful, not only for raising their grades but also for enhancing their learning. Most survey respondents did not need to remediate an exam but still perceived benefits from knowing the option was available, describing lower stress levels and a more conducive learning environment as positive aspects. Conclusion Incorporation of this structured remediation process into a veterinary immunology course promoted student learning, facilitated communication with struggling students, and contributed to a positive learning environment. Funding Source N/A Topic Categories Immunology Education and Communication (EDU)
Acquired Digital Fibrokeratoma Presenting In a Patient With WHIM (Warts, Hypogammaglobulinemia, Infections, Myelokathexis) Syndrome 2333706
Abstract Introduction WHIM syndrome is a primary immunodeficiency disorder with an estimated prevalence of 1 in 5 million live births and less than 100 documented cases in the medical literature. It’s an autosomal dominant mutation of the CXCR4 gene. The gain-of-function mutation results in aberrant CXCL12—CXCR4 signaling and impaired neutrophil egress from the bone marrow. Clinical manifestations include recurrent bacterial infections, hypogammaglobulinemia, neutropenia, and increased risk of HPV infection. Cutaneous findings in WHIM syndrome are generally attributed to HPV associated conditions. Methods Case: A 60 y/o woman presented with a firm, hyperkeratotic, pedunculated papule on the left middle finger. Given the patient’s history of HPV-related disease, the lesion was initially suspected to be a verruca or squamous cell carcinoma. Histopathologic evaluation demonstrated epidermal hyperkeratosis overlying a core of dense, vertically oriented collagen bundles with sparse fibroblasts, consistent with an acquired digital fibrokeratoma. There was no histologic evidence of viral cytopathic effect or HPV-associated changes. The lesion was fully excised with no recurrence. Results Acquired digital fibrokeratoma is a rare benign fibrous tumor of unclear pathogenesis, thought to arise from localized trauma or dysregulated fibroblast activity. In the context of WHIM syndrome, chronic immune dysregulation, altered chemokine signaling, and impaired healing in addition to trauma may contribute to aberrant fibrous proliferation. This highlights that not all cutaneous lesions in WHIM syndrome are infectious or virally mediated. Conclusion As the first documented case of acquired digital fibrokeratoma in a patient with WHIM syndrome, it expands the recognized phenotypic spectrum of WHIM-associated cutaneous manifestations and suggests a potential link between immune dysregulation and noninfectious fibrous tumor development. Awareness of this may aid clinicians in diagnostic evaluation and management. Funding Source n/a Topic Categories Cytokines and Chemokines and their Receptors (CCR)
Nuclear Spin Relaxation Modulation of Fluorinated Deep Eutectic Solvents for Radiofrequency Information Encoding and Display
ABSTRACT Nuclear spin properties underpin nuclear magnetic resonance. The tunability of 19 F spin systems renders fluorinated materials versatile platforms for diverse applications. Although frequency encoding of 19 F nuclei enables high‐density molecular encoding, the use of spin relaxation remains limited. Here, a 19 F magnetic resonance display platform that exploits spin relaxation modulation through synergistic material‐level design and instrument‐level pulse sequence control is presented. Incorporation of paramagnetic metal ions into fluorinated deep eutectic solvents enables precise tuning of 19 F relaxation times for information encoding. Tailored relaxation‐selective pulse sequences allow selective visualization of the encoded information. Iron‐containing fluorinated deep eutectic solvents further enable reversible data editing via photoredox responses. In addition, UV‐printable paramagnetic fluorinated ionogels allow rapid and rewritable information display. This work expands nuclear magnetic information encoding by introducing controllable 19 F spin relaxation as an additional information domain, thereby converting spin relaxation from a physical parameter into an information carrier and enabling a selective, responsive, and non‐luminescent display platform well suited for high‐confidential anticounterfeiting and information display.
Volumetric comparison of brain structures in early and advanced bilateral glaucoma patients using 7 tesla MRI
Abstract Glaucoma is a progressive neurodegenerative disorder characterized by retinal ganglion cell loss and corresponding visual field (VF) impairment. Although traditionally viewed as an ocular disease, accumulating evidence suggests involvement of central nervous system structures, particularly the lateral geniculate nucleus (LGN). In this cross-sectional study, we enrolled 67 participants: 25 with bilateral early-stage glaucoma, 27 with bilateral advanced-stage glaucoma, and 15 age-matched healthy controls. All participants underwent 7‑Tesla magnetic resonance imaging, including high-resolution 3D T1‑weighted and magnetization transfer‑weighted SILENT sequences. LGN volumes were manually segmented, and whole‑brain volumetry was assessed with FreeSurfer. Compared with both early glaucoma patients and healthy controls, advanced glaucoma patients presented significantly reduced LGN volumes. LGN volume was positively correlated with retinal nerve fiber layer thickness, particularly in the superior and inferior quadrants, and decreased with increasing visual field loss expressed as mean deviation (MD). The correlation between LGN volume and MD was stronger for the right side than for the left side. No significant volumetric differences were observed in other major visual or brain regions across groups. These findings support structural involvement of the central visual pathway in glaucoma and suggest that LGN volume may reflect disease severity, particularly in advanced stages. At present, 7 Tesla MRI should be considered primarily as a research tool that may complement understanding of glaucoma related visual pathway changes.
Discovery and delineation of human antibody-secreting cell subsets 2256534
Abstract Introduction Antibody-secreting cells (ASCs) differentiate from B cells after immune activation and home to inflamed tissues or to sites which harbor protective niches, such as bone marrow (BM). ASCs have both salutary and pathogenic roles across various disease contexts. Their lifespans range from days to decades, and they exhibit diverse functionality including antibody secretion, cytokine production, and immunoregulation. Currently, we cannot identify, ascribe function to, nor manipulate distinct human ASC subsets, preventing targeted therapeutic intervention and selective induction. Methods To discover ASC subsets, we applied multi-omic single-cell sequencing to healthy human BM and peripheral blood, quantifying surface protein expression, transcriptomics, and chromatin accessibility. To evaluate ASC localization and cellular niches, we performed highly-multiplexed spatial proteomics on BM and splenic biopsies from healthy adults. We developed a simple gating strategy to prospectively isolate ASC subsets. We cultured sorted ASC subsets from healthy adult BM and evaluated specificity to antigens induced in childhood vaccination. Results We discovered five ASC subsets with distinct molecular features. We found significant differences in the expression of cell adhesion, cell cycle, and metabolic programs across ASC subsets. We characterized cell morphology and niches across subsets and tissues. We observed specificity to antigens from childhood vaccination in several ASC subsets. Conclusion Combining a systems immunology approach with functional assays revealed unappreciated ASC subsets and provided the highest resolution assessment of human ASC identity reported to date. We identified gene programs that are differentially expressed as ASCs exit the blood and transition into BM-resident cells. We found that durable immune memory is not harbored by a single ASC subset. Our findings enhance the identification and monitoring of ASCs and offer the promise of therapeutic manipulation in clinical settings. Funding Source The Paul G. Allen Family Foundation Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)
Characterizing the role of ovarian hormones in regulating sex differences in baseline immune parameters 2309498
Abstract Introduction Sex differences among peripheral blood mononuclear cells (PBMCs) have been previously described in humans, including greater percentages (%) of CD4+ T cells in females (XX) and greater % of natural killer cells in males (XY). The role of the ovaries in regulating these and other sex differences is unknown. Methods We collected PBMCs from 80 age-matched participants (reported as mean ± SEM) and stratified into control females (n = 28; 41.6 ± 1.0 years), control males (n = 20; 40.4 ± 1.3 years), bilateral salpingo-oophorectomy (BSO) recipients (n = 16; 46.1 ± 2.0 years), and BSO recipients receiving hormone therapy (HT) (n = 16; 45.7 ± 1.2). BSO recipients were at least 1 year out from surgery. PBMCs were isolated to analyze subsets by single-cell mass cytometry. Serum was collected for estradiol (E2), testosterone (T), and progesterone measurements, and cytokines were quantified using the O-LINK platform. Results Three serum markers (IL-10, IL-17c, and CXCL11) showed a sex difference, with greater levels in males; no cytokine was modulated by BSO. Among CD45+ PBMCs, females had a greater % of CD3+ T cells, whereas the % of monocytes were greater in males. However, the sex difference in monocytes disappeared when accounting for T cell differences. Within CD4+ T cells, males had higher % of regulatory T cells and circulating follicular T cells, while within CD8+ T cells, they had higher percentages of PD-1+ effector and terminally differentiated subsets. No highlighted sex difference was associated with the presence of the ovaries. Further explorations identified a negative correlation of % of CD3+ T cells with T levels. Though not driving the sex differences, BSO did increase the % of naïve B cells at the expense of memory B cells, and this was reversed with HT. The % of naïve B cells negatively correlated with E2 and T levels. Conclusion The ovaries are not a major regulator of sex differences in immune subsets but do repress the generation of naïve B cells. Funding Source CIHR - Canada Graduate Scholarship Topic Categories Immune Mechanisms of Human Disease (HUM)
The role of the T-cell receptor (TCR) in the generation of lung CD4+ tissue-resident memory T cells (TRM) following influenza infection 2307595
Abstract Introduction Upon initial antigen encounter, anti-influenza T cells infiltrate the lung and form tissue-resident memory T cells (TRM), that provide a rapid response to antigen re-encounter. While the mechanisms driving lung CD8+ TRM formation are well defined, CD4+ TRM are less understood. Methods In order to define the role of the TCR in the formation of CD4+ TRM in the lung after influenza infection, we employed a novel TCR transgenic mouse line, GPB1, that expresses a fixed TCRB chain derived from a TCR specific for the MHC Class II-restricted epitope of the LCMV glycoprotein, GP61-80, paired to a polyclonal TCRa repertoire, crossed to a mouse expressing a Nur77-GFP reporter to monitor TCR signal strength. We transferred GPB1 T cells into WT hosts followed by a primary or heterologous secondary challenge with recombinant PR8 expressing GP61-80 (PR8-gp61), enabling us to track T cells across a broad range of TCR avidity and signal strength. Results Lung-infiltrating effector CD4+ T cells displayed higher TCR avidity and enhanced TCR signal strength compared to those in the mediastinal lymph nodes (medLN). This was true for both primary PR8-gp61 infection as well as heterologous secondary PR8-gp61 challenge biased towards Tfh-like (due to prior protein immunization with recombinant GP) or Th1-like (due to prior LCMV infection) effector responses in the lung. Resulting lung CD4+ TRM also skewed towards high avidity as compared to medLN. Enhanced TCR avidity and signal strength in the lung was associated with induced expression of the integrin CD49b and the chemokine receptor CXCR6. Conclusion We conclude that strong TCR signaling in high avidity CD4+ T cells drives lung-homing effector T cell responses and establishment of TRM. Future work will establish the mechanistic link between TCR signal strength and the upregulation of molecules required for homing to or retention within the lung. Funding Source R01AI194555, T32AI138945 Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)
Tumor Egfl6 is a new driver of neutrophil extracellular traps (NETs) in ovarian cancer 2310080
Abstract Introduction Ovarian cancer (OvCa) is characterized by immunosuppressive tumor microenvironment (TME). We displayed the tumor-secreted factor EGF-like domain multiple-6 (EGFL6) drives an immunosuppressive myeloid phenotype, inducing CXCL2, IL-8, and other factors associated with neutrophil extracellular trap (NET) formation. Our preliminary data showed in Egfl6-expressing ID8p53-/- tumors NET infiltration increased compared to parental tumors. Here, we evaluated the role of EGFL6 as a regulator of NETosis in the OvCa TME and whether pharmacological NETosis blockade can reverse Egfl6-induced immunosuppression. Methods In vitro assays were used to evaluate the direct role of recombinant Egfl6 (rEgfl6) on NETosis. OvCa patient tissues were analyzed by immunostaining and qPCR to assess EGFL6 expression, immune cell abundance, and NET markers, such as Citrullinated histone 3 (Cit H3) and neutrophil elastase (NE). To test whether NETosis inhibition reprograms the EGFL6-induced immunosuppression, Egfl6+/- ID8p53-/- tumor-bearing mice were treated with GSK484, a selective PAD4 inhibitor. Immune profile analyzed within murine ascites and tumors via flow cytometry. Results rEgfl6 directly enhanced NETosis in vitro. OvCa patient tissues showed high EGFL6 and NE expression, and NK cells were frequently localized near CitH3+ NETs. In vivo, GSK484 treatment increased cytotoxic CD107+ NK cells and reduced expression of inhibitory receptors including NKG2A and KLRG1. In myeloid cells, GSK484 upregulated antigen-presenting markers (MHCII, CD86) and reduced infiltration of Tregs in Egfl6+ TME. Conclusion Our findings identify EGFL6 as a previously unrecognized tumor-derived driver of NETosis, contributes to immunosuppression in OvCa. NETs directly impair NK cell functions in Egfl6+ tumors, while NETosis inhibition with GSK484 partially restores anti-tumor immunity by enhancing NK cell cytotoxic and reprogramming myeloid cells. These data support targeting NETosis as a therapeutic approach to enhance current therapies in OvCa. Funding Source NIH Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
Discovery of novel antifungal candidates for coccidioidomycosis through integrated drug screening and proteomics 2261274
Abstract Introduction Inhalation of airborne Coccidioides species causes coccidioidomycosis, a severe respiratory disease ranked as a significant pathogen by WHO. With infections reaching new records each year, efforts to develop efficient treatments with minimal adverse reactions have been ineffective. This study addresses these gaps by integrating computational and experimental methodologies to discover novel Coccidioides therapeutics and virulence factors. Methods We used computational pipelines to classify 45 Coccidioides-inhibitory chemicals and screened DrugBank and ChEMBL databases based on the chemical grouping of these chemicals to identify drugs for repurposing. In parallel, leveraging California’s botanical diversity, we developed an experimental screening protocol using colorimetric assays and colony forming unit plating to test antifungal drug candidates. To support these drug discovery efforts, we functionally annotated C. immitis hypothetical protein sequences using bioinformatics tools. Comparative analysis with known respiratory fungal virulence factors identified proteases for structural homology analysis and prediction of host immune pathway interactions. Results We revealed ∼9,000 high-quality proteins and identified 300 unannotated proteases as potential therapeutic targets. The experimental screening methodology revealed that locally grown crops provide potent hits, demonstrating the importance of applying local resources to tackle regional problems. Conclusion These approaches provide a foundation for developing improved coccidioidomycosis treatments. Funding Source NIH R15AI174173, NIH T32-GM141862, GEM consortium fellowship Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)
Identification of Coccidioides specific T cell clones and antigens in naturally exposed Pig-tailed Macaques and Valley fever patients 2308943
Abstract Introduction Coccidioidomycosis (Valley fever (VF)) is frequently misdiagnosed and mistreated as bacterial or viral pneumonia due to similar clinical presentations. Severe VF manifests in < 5% of symptomatic cases and can be life-threatening. T cell responses play a vital role in vaccine-induced protection in mouse models and appear to be critical for resolution of infection in humans. We set out to identify T cell antigens and responses that are generated in pig-tailed macaques (PTM) and humans to aid antigen discovery and model development. Methods Peripheral blood mononuclear cells (PBMCs) were collected from both humans and PTM. The PBMCs were non-specifically expanded then stimulated with overlapping pools of Coccidioides-specific peptides from 27 antigens following an antigen multiplexing scheme. Activated T cells were sorted, T cell receptor (TCR) sequenced, and the TCR clonotypes were associated with the stimulating Coccidioides antigens. Specific T cell responses were determined by ELISPOT or single cell sequencing. Results Our results have confirmed reactivity of previously known Coccidioides antigens while also identifying new antigens not previously associated with T cell immunogenicity. T cell responses varied between animals and humans, but similar antigens were recognized. Th17 and Th1 immune responses and responding clones were identified by single cells sequencing. Conclusion Similar responses were seen in PTM and humans, supporting the PTM model for VF research. Some identified T cell clones recognized antigens that have been associated with a mouse protective DNA vaccination. These antigens and their related clones will be tracked in challenged or vaccinated PTM to aid in the identification of correlates of protection. Finally, the reactive antigens and their epitopes can be further investigated for potential diagnostic tools or vaccines. Funding Source U19AI166058, 3U19AI166058-02S1, P51OD010425, U42OD011123 Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)
Immune alteration in the CNS tissues during peripheral influenza infection 2333891
Abstract Introduction Growing evidence suggests that peripheral infections contribute to neurodegenerative disease progression and cognitive impairment, yet the underlying neuro-immune mechanisms remain poorly understood. Methods Here, we used a mouse model of Influenza A virus (H1N1) respiratory infection to investigate immune responses in the central nervous system (CNS). Results At day 6 post-infection, we observed significant increases in microglia, perivascular macrophages, monocytes, NK cells, and neutrophils, indicating infection-driven immune activation within the CNS. Strikingly, eosinophils persisted in both brain/leptomeninges and dura mater up to 7 weeks post-infection. No viral RNA was detected in the brain at any time point, whereas lung tissue harbored high viral loads, suggesting peripheral infection as the driver of neuroinflammation. Cytokine and chemokine profiling revealed persistent elevation of CCL11 in serum at both day 6 and week 7. CCL11 has been implicated in other peripheral infection related neuroinflammation, potentially contributing to myelin loss and impaired neurogenesis, although mechanisms remain unknown. To address the role of CCL11 in CNS inflammation and dysfunction in this context, we have begun characterizing CCL11 knockout (KO) mice following influenza infection. Preliminary results indicate that CCL11-KO mice are more susceptible to infection compared to controls, highlighting a complex role for CCL11 in balancing antiviral defense and neuroinflammation. Furthermore, scRNA-seq analysis of healthy mice revealed high CCL11 expression in dural sinus smooth muscle cells, leptomeningeal fibroblasts and brain pericytes suggesting that brain border cells may contribute to local regulation of immune responses after infection. Conclusion Moving forward, our goal is to investigate CCL11 as a potential mediator linking peripheral respiratory infection to persistent neuroinflammation, providing insight into mechanisms by which infections may contribute to long-term brain dysfunction. Funding Source Emerging pathogen consortia, HHMI Topic Categories Neuroimmunology (NEUR)
Tantalum‐Based Nanomaterials: From Properties to Biomedical Applications
ABSTRACT For decades, tantalum has been a cornerstone of surgical implants, highly favored for its excellent chemical stability and biocompatibility. With the emergence of nanotechnology, tantalum‐based nanomaterials (TBNs) have become highly promising candidates in biomedical applications due to their adjustable physicochemical and biological properties. However, existing reviews are mostly limited to a single material type or specific application direction, lacking a systematic integration of various TBNs and their diverse biomedical applications. This review systematically summarizes the latest research progress on TBNs. Starting from their performance advantages, it further reveals their significant applications in biomedical applications, including biosensing, medical imaging, tissue engineering, anti‐tumor therapy, embolization, and radiation protection. Finally, it summarizes the current challenges and prospects for future development directions, providing an important reference for promoting the clinical translation of TBNs and the design of integrated diagnosis and treatment platforms.
Positively Charged Polymer‐Brush MOFs for Large‐Area, Pressure‐Resistant Gas Separation Membranes
ABSTRACT Scalable fabrication of high‐performance gas separation membranes remains a major challenge for energy‐efficient gas purification. Industrial translation of pressure‐resistant mixed matrix membranes (MMMs) is largely impeded by the dispersion instability of nanofillers under rapid, non‐equilibrium manufacturing conditions, which leads to uncontrollable aggregation and interfacial defects during processing. Here, we present a universal “pre‐occupation and post‐activation” strategy to construct positively charged polymer‐brush metal–organic frameworks (MOFs). This approach endows the fillers with a dual‐stabilization mechanism: electro‐steric effects ensure static dispersion stability, while a hydrogen‐bonding‐driven interfacial interlocking mechanism maintains stability during dynamic processing. Together, these mechanisms synergistically facilitate the seamless integration of nanofillers within ultrathin selective layers. This design allows roll‐to‐roll production of 1 m‐wide, pressure‐resistant mixed matrix composite membranes (MMCMs) with tunable CO 2 separation performance. The resulting MMCMs deliver outstanding CO 2 /CH 4 and CO 2 /N 2 separation under industrially relevant pressures, reducing the required membrane area by more than an order of magnitude compared with laboratory‐scale membranes. A spiral‐wound module with an effective area of 0.4–2 m 2 further validates the scalability and operational robustness. This work overcomes a long‐standing barrier in MMM processing, marking a significant step toward industrial implementation of MOF‐based, energy‐efficient gas separation technologies.
A novel CD69 binding ankyrin repeat protein (Ankyron®) enables detection of early activated and antigen-specific T cells in pigs
Abstract T cells are essential for a wide range of protective immune responses, but significant gaps remain in our understanding of their phenotypic and functional diversity, especially in large animal models, like the pig. This is partly due to the limited availability of porcine-specific reagents and validated markers to study T-cell activation and differentiation. One marker involved in both of these processes is CD69: In mice and humans, it serves as an early activation marker and is used to define tissue-resident memory T cells. However, its use for the identification and functional assessment of T-cell subsets in pigs has been previously restricted by the lack of monoclonal antibodies. To address this limitation, we evaluated an alternative approach to antibodies: A novel ankyrin repeat protein, Ankyron ® EG40360, was used as a tool to detect porcine CD69. Ankyrin repeat proteins offer advantages over conventional antibodies, including their small size and simplified integration into flow cytometry panels due to the absence of isotype restrictions. We confirmed specific binding of Ankyron EG40360 to porcine CD69 using both flow cytometry and microscopy in porcine CD69-transfected HEK293T cells. Thereafter, we studied CD69 expression in PBMCs stimulated with PMA/ionomycin. While CD69 was basically absent in non-stimulated control cells, CD69 was highly expressed upon PMA/ionomycin stimulation as early as 4 h post stimulation. Thereby, we could confirm CD69 as an early marker of lymphocyte activation in pigs. To further assess the expression of CD69 with the Ankyron EG40360 in a more natural setting, we studied CD69 expression in PBMCs from pigs that received a protein subunit vaccine. Following in vitro restimulation with the vaccine antigen, the frequency of CD4 and CD8 T cells expressing CD69 and CD25 increased indicating detection of antigen-specific T cells and making CD69 a potential target for activation-induced marker assays in pigs. Thus, Ankyron EG40360 expands the porcine immunology toolkit and will facilitate a more comprehensive characterization of activated T cells in vaccine development and infection studies.
Guanylate-Binding Protein 5 (GBP5) Exacerbates Sepsis by Inhibiting STING-mediated Post-Golgi Transport 2256813
Abstract Introduction Excessive activation of STING is associated with sepsis. Maintaining appropriate self-limiting mechanisms following STING activation is crucial. Upon activation, STING in the Golgi can be degraded via the lysosomal-autophagic pathway or recruited to COP I vesicles for retrograde transport back to the ER. GBP5 is localized to the Golgi and plays a crucial role in the immune response. However, its potential functional relationship with STING in sepsis remains unclear. Methods The role of GBP5 in sepsis was investigated by assessing inflammation and intestinal injury severity levels in WT/GBP5 KO mice. Subsequently, molecular docking and immunoprecipitation (IP) experiments identified a potential interaction between GBP5 and STING. Cellular experiments further evaluated the specific mechanism by which GBP5 inhibits STING-mediated post-Golgi transport. Results Elevated GBP5 expression was observed in sepsis patients, correlating positively with STING levels. Overexpression of GBP5 enhanced STING activation, while GBP5 KO inhibited STING phosphorylation. GBP5 interacts with STING via its ligand-binding domain, and this interaction is associated with the localization of GBP5 in the Golgi. Mass spectrometry analysis showed that GBP5 is associated with vesicle transport in the Golgi and autophagosome assembly. Following STING activation, co-localization between STING and the Golgi apparatus was significantly reduced in the GBP5 KO group. GBP5 overexpression inhibited autophagy, and in the GBP5 KO group, STING levels were reduced in the ER but elevated in the Golgi. Survival and intestinal injury outcomes were significantly improved in GBP5 KO mice following CLP. Conclusion GBP5 interacts with activated STING at the Golgi, exerting dual regulatory effects: preventing autophagic degradation of STING and inhibiting its retrograde transport to the ER. This enhances STING signaling, thereby exacerbating the inflammatory response and tissue damage caused by sepsis. Funding Source n/a Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)
Decoupling Drug Exposure, Binding ADA, and Functional Neutralization in Anti—PD-L1 Therapy Using Orthogonal ELISA Immunogenicity Assessment 2309558
Abstract Introduction Immune checkpoint inhibitors targeting PD-L1 can trigger anti-drug antibody (ADA) responses with varied biological effects. In anti-PD-L1 therapy, circulating drug levels and binding ADA detected via immunogenicity screening do not reliably predict loss of immune checkpoint blockade. Functional neutralizing antibodies (NAbs), which directly block PD-1/PD-L1 signaling, provide a distinct, biologically relevant immunogenicity measure. Accurate assessment thus demands separate evaluation of drug exposure, immunogenicity incidence, and functional neutralization. Methods We applied an ICH-aligned, tiered ELISA-based framework to to evaluate anti-PD-L1 therapy in human serum samples containing Pembrolizumab. Tier 1 used a sandwich therapeutic drug monitoring (TDM) ELISA to quantify total circulating drug. Tier 2 screened for binding ADA via drug-bridging ELISA. Tier 3 assessed NAb activity with a competitive receptor-blocking ELISA measuring PD-L1/PD-1 interaction inhibition. All assays were validated for sensitivity, specificity, precision, and drug tolerance. Results Binding ADA often appeared without loss of PD-L1 pathway inhibition, preserving pharmacological activity despite immunogenicity. Conversely, Tier 3 NAb-positive samples showed reduced PD-L1 blockade, even with detectable Pembrolizumab, confirming functional neutralization. These findings highlight drug exposure, binding ADA, and neutralizing activity as non-redundant variables in anti-PD-L1 therapy. Conclusion Overall, tiered, functionally informed immunogenicity assessment is essential for interpreting responses to anti-PD-L1 therapy. Integrating TDM, binding ADA detection, and NAb ELISAs distinguishes exposure, immunogenicity, and PD-1/PD-L1 blockade loss, enhancing mechanistic insights and risk assessment for checkpoint inhibitors. Funding Source n/a Topic Categories Translational and Interventional Immunology (TI)
Bioactive Enhanced Adjuvant Chemokine Oligonucleotide Nanoparticles (BEACON) for Mucosal Vaccination Against Genital Herpes 2307637
Abstract Introduction Genital herpes (caused primarily by herpes simplex virus 2 [HSV-2]) remains highly prevalent with no licensed vaccine, in part because intramuscular vaccines are unable to generate robust induction of vaginal tissue-resident memory T cells (Trm) and mucosal antibodies. Our previous strategy of “Prime and Pull”, namely, priming with a conventional intramuscular vaccine followed by vaginal chemokine application to pull effector T cells, confers protection against disease but not infection. Thus, we developed Bioactive Enhanced Adjuvant Chemokine Oligonucleotide Nanoparticles (BEACON) as a mucosal adjuvant to promote vaginal immunity and reduce viral burden and disease. Methods CXCL9 and CpG DNA were electrostatically assembled into nanoparticles to create BEACON, and assessed in vitro for chemotaxis and innate immune activation. Female mice were primed intramuscularly (i.m.) with HSV glycoprotein D (gD) mRNA-LNPs and boosted i.m. or intravaginally with gD and BEACON. Vaginal inflammation and CpG uptake, Trm, gD-specific serum and vaginal IgG/IgA, viral shedding/dorsal root ganglion (DRG) viral DNA, and survival after lethal genital HSV-2 challenge were quantified. Results Compared with free CpG, BEACON increased CpG uptake by vaginal APCs (∼10-fold) and reduced neutrophil uptake and local inflammation. Intravaginal glycoprotein plus BEACON given to mice previously immunized with mRNA vaccine uniquely expanded vaginal HSV-specific CD8+ Trm and induced high local IgG and IgA. This regimen provided complete protection against lethal disease, accelerated viral shedding control (significant by day 2), and reduced DRG viral genomes (∼80-fold) compared with intramuscular boosting. Conclusion An intramuscular prime followed by BEACON-enabled mucosal boost safely coordinates Trm and mucosal antibody and markedly improves protection against genital HSV-2, supporting a generalizable mucosal boosting strategy for HSV-2 and other sexually transmitted infections. Funding Source HHMI Topic Categories Vaccines and Immunotherapy (VAC)
Fibroblasts as Protective Immunoregulators in Oral Mucosal Disease 2310061
Abstract Introduction The oral cavity is a critical barrier site that is continuously exposed to a variety of stimuli, including dietary components, the microbiome, environmental antigens, and mechanical stress and damage. Dysbiosis in this environment can lead to mucosal inflammation and subsequent alveolar bone destruction and tooth loss, a condition known as periodontitis. Our lab has previously highlighted the contribution of hematopoietic cells, especially Th17 cells and neutrophils, to the pathogenesis of periodontitis. However, insights from a human oral mucosal scRNA-seq atlas suggested that mesenchymal cells, including fibroblasts, may actively play a role during inflammation in the oral cavity.Based on this observation, we hypothesized that mesenchymal cells, particularly fibroblasts, contribute to oral mucosal pathogenesis. Methods Utilizing a mouse ligature-induced periodontitis (LIP) model which recaptures key features of human periodontitis, including dysbiosis, bone destruction, and immune cell infiltration, we performed time-course scRNA-seq to examine the transcriptome profile across different cell types in the oral mucosa during periodontitis development. Results Our analysis identified a subset of fibroblasts that became rapidly activated within 12 hours after ligature placement, exhibiting gene expression signatures associated with tissue inflammation and damage-related response, implicating the IL-33—ST2 signaling axis. The activated fibroblasts produced soluble ST2 (sST2), a decoy receptor for IL-33, early after the ligature placement. Functional studies in vivo using sST2 KO mice demonstrated sST2 KO mice significantly increased alveolar bone destruction compared to WT controls in the LIP model, indicating a protective role for sST2 in oral mucosal inflammation. Conclusion Overall, our work highlights the critical function of fibroblasts as immune-modulatory stromal cells that sense tissue stress and orchestrate inflammatory responses in the oral mucosa. Funding Source This project is funded by the National Institute of Allergy and Infectious Diseases and the National Institute of Dental and Craniofacial Research intramural program. Topic Categories Mucosal and Regional Immunology (MUC)