Browse Articles

Nanoparticle delivery amplifies the adjuvant effects of TLR agonists for cancer vaccines 2310294

Abstract Introduction Nanoparticle delivery systems are widely used to improve vaccine adjuvant effects. IVAX, a nanoparticulate adjuvant system encapsulating a combination of Toll-like receptor (TLR) 4 and TLR9 agonists, has shown significant promise in various vaccine models. However, the role of nanoparticles in the adjuvant system was not well-defined. This study aims to investigate how different nanoparticle carriers affect the immunogenic performance of adjuvanted vaccine formulations in a melanoma vaccine model. Methods Three representative nanoparticles, including nanoemulsion (IVAX-1), cationic lipid nanoparticle (LNP; IVAX-5), and ionizable LNP (IVAX-6), were formulated to encapsulate a combination adjuvant of CpG and MPLA using ovalbumin (OVA) as a model antigen. Physicochemical properties of all three nanoparticles were characterized, followed by the evaluation of NF-κB activation using the RAW-Blue reporter assay. Furthermore, humoral and cellular immune responses, reactogenicity, and antitumor efficacy across vaccine formulations were evaluated in B16F10-OVA melanoma models. Results Nanoparticle-formulated TLR agonist adjuvants enhanced NF-κB activation compared to controls, indicating a nanoparticle-dependent effect on innate immune signaling. Mice immunized with adjuvanted vaccines showed complete tumor protection through the study endpoint, whereas buffer-treated and antigen-only control groups developed progressive tumor growth. Immunoprofiling post-vaccination revealed the differences in immune responses induced by different nanoparticle formulations. Conclusion Together, this study highlights the importance of nanoparticle carrier design in shaping the performance of TLR-agonist adjuvanted cancer vaccines. Funding Source This research was supported by the National Institute of Allergy and Infectious Diseases (NIAID) of the National Institutes of Health (NIH) under Award Number 1U19AI181968-01 Topic Categories Vaccines and Immunotherapy (VAC)

Data‐Driven Design of Self‐Assembled Monolayers for High‐Efficiency Perovskite Solar Cells

ABSTRACT Self‐assembled monolayers (SAMs) are pivotal for boosting the performance of perovskite solar cells (PSCs). Yet, the intricate link between molecular structure and device efficiency hinders rational SAM design. Here, we introduce a data‐driven strategy that leverages a curated dataset of reported SAMs and their PSC efficiencies, with molecular structures encoded into three distinct segments: anchor group–linker–head group. Based on this fragment‐encoding framework, our strategy focuses on the recombination of fragment units, rather than unconstrained de novo molecular design. Using ensemble learning and SHapley Additive exPlanations (SHAP) interpretability within a cross‐validated framework, we pinpointed the head group as the dominant performance driver. This insight guided the construction of an expanded molecular library by recombining high‐value fragments identified from the curated database. Virtual screening of this library then yielded a synthetically accessible SAM molecule with top‐predicted efficiency, namely S1. Experimental validation revealed that S1 forms a compact, ordered monolayer on NiO x , featuring a well‐aligned HOMO level and a strong interfacial dipole that optimizes electronic coupling. Consequently, S1 enables defect passivation and hole extraction, delivering a champion power conversion efficiency of 26.21%. This study establishes a machine learning paradigm, integrating fragment‐based encoding and explainable AI for data‐driven interface optimization in high‐performance PSCs.

Decoupling Parasitic Reactions From Bravais Law‐Guided Electroredox Toward Highly Reversible (101)‐Textured Zn Anodes for Ah‐Scale Batteries

ABSTRACT Aqueous Zn–ion batteries (AZIBs) are promising for grid‐scale energy storage but are limited by poor Zn anode reversibility due to dendrite growth and water‐driven parasitic reactions. Although crystallographic texture regulation based on Bravais law can guide Zn plating/stripping, selective facet screening often leaves unprotected facets vulnerable to the parasitic side reactions. This inherent trade‐off in conventional Bravais law‐based texturing strategies leads to unstable and transient texture evolution especially under practical conditions. In this study, we propose a decoupled electrolyte design that simultaneously enables facet‐selective texture control and global suppression of water activity using a formamide (FA) cosolvent and a trace 1‐butyl‐3‐methylimidazolium cation (Bmim + ) additive. Bmim + additive preferentially adsorbs on the Zn(101) facet, retarding its growth and directing Zn plating/stripping toward a (101)‐textured mode, while FA suppresses the bulk/interfacial water activity, thereby suppressing interfacial side reactions on non‐targeted facets. This hierarchical design ensures sustained Zn(101)‐textured electroredox with markedly improved reversibility, delivering 1700 h lifespan in Zn||Zn symmetric cells at 5 mA cm −2 , 5 mAh cm −2 , and 5000 cycles in Zn||I 2 full cells with 79.55% capacity retention at 0.5 A g −1 . Notably, a 1.4 Ah pouch cell further validates the scalability of the proposed decoupling principle for practical AZIBs.

Exosomal misfolded α1-antitrypsin triggers a cytosolic GRP78-dependent unfolded protein response and pro-survival signaling in pre-metastatic tissues

Inducing transcription in a gene desert is associated with locally increased replication activity

The role of the transcription factor Blimp-1 in T cells in anti-PD1 immunotherapy in non-small cell lung cancer 2258857

Abstract Introduction Immune checkpoint inhibitors targeting the interaction between PD1 and PDL1 are effective for immunotherapy in non-small cell lung cancer (NSCLC), which is the leading cause of cancer related death in the world. However, only subgroups of patients respond to therapy suggesting the existence of resistance mechanisms. Blimp-1 is a transcription repressor that interacting with other transcription factors in lymphocytes regulate their cellular fate. Methods In this study,we analyzed post-surgery lung tissues and peripheral blood cells from patients with Non Small Cell Lung Cancer (NSCLC) and control subjects by using western blot, immunohistochemistry, ELISA, multiplex cytokine and qPCR analysis. Moreover, we analyzed in a murine model of NSCLC, the role of Blimp1 in T cells after targeting of Blimp-1 in T cells expressing lymphocyte-specific-protein tyrosine kinase (Lck), a kinase crucial for T cell receptor signaling. In addition we blocked PD1 in this murine model of lung cancer. Results Here, we found progressive increased Blimp1 expression in the tumoral region of NSCLC patients. In PBMCs, Blimp1 was expressed in CD8+ T cells but predominantly in immunosuppressive Foxp3+ Treg cells. Targeting PD1 in PBMCs reduced Blimp1. In a murine model of NSCLC, T cell targeting of Blimp-1 resulted in reduced tumor load, suppressed lung Foxp3+ Treg cells and induced T-bet+ T effector cells producing IFN-gamma. Furthermore, RNA sequencing of T cells showed an induction of Th1 and Tc1 markers and PD1 in the absence of Blimp1. Targeting PD1 on a Blimp-1 deficiency in T cells in tumor resulted in induction of cytotoxic and effector T cells. Conclusion These data indicate that the tumor microenvironment induces Blimp-1 in immunosuppressive Treg and CD8+ T effector cells, thereby limiting the therapeutic efficacy of anti-tumor immune responses. Targeting of Blimp-1 emerges as a novel concept to suppress immune evasion during anti-PD1 immunotherapy in lung cancer. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

High fat diet drives rapid production of donor specific antibody in heart transplantation 2307505

Abstract Introduction Solid organ transplantation is the most effective curative treatment for end-stage organ failure and while immunosuppression regimens have drastically improved graft survival by blocking acute cellular rejection, both acute and chronic antibody mediated rejection (ABMR) remain key mediators of graft loss. Donor specific antibody (DSA) is produced by plasma cells (PC) generated through germinal center (GC)-dependent or independent mechanisms. We hypothesized that in obesity GC-independent or extrafollicular (EF) pathways are increased leading to more rapid production of DSA. Methods C57BL/6 mice received HFD or control diet for four weeks prior to receiving Balb/c heart allografts. DSA was measured by crossmatch assay of serum with donor thymocytes. Animals were sacrificed at 5- and 14-days post transplantation. Splenic immune cell composition and function was analyzed by flow cytometry and immunohistochemistry. Graft infiltrating cells and complement deposition were measured by histology. Results Relative to control diet, mice on HFD had increased B cell activation at day 5 post-transplant and an increase in serum DSA with canonically EF IgG subtypes compared with canonically GC-dependent subtypes. At 14 days post-transplant mice on HFD had markedly increased total and donor specific splenic PCs which roughly correlated to their weight. Spleens of HFD mice had no increase in GC B cells and fewer follicular helper T cells. Therefore, we inferred that high fat diet enhances DSA PC generation through GC-independent or EF pathways. Conclusion DSA from EF PCs is less dependent on T cell derived signals and therefore less sensitive to traditional immunosuppression. We observed that PCs generated in response to transplant and immunization in mice on HFD have distinct metabolic profiles including increased oxidative phosphorylation and fatty acid oxidation and mitochondrial activity. These data suggests that targeting PC metabolism therapeutically or prophylactically will reduce DSA and ameliorate ABMR. Funding Source n/a Topic Categories Transplantation Immunology (TRAN)

Leveraging the adaptive NKG2C-HLA-E axis using neural network identified peptides for HER2-targeted NK cell immunotherapy 2334737

Abstract Introduction Breast cancer remains a leading cause of cancer-related mortality globally, with HER2+ tumors representing 15-20% of cases further complicated by treatment resistance. As a novel treatment approach, natural killer (NK) cells are ideal antitumoral effectors that circumvent MHC restriction and T-cell therapy associated toxicities. Although recognized as innate effectors, subsets of NK cells called adaptive NK (aNK) cells show antigen-specific memory responses via the NKG2C-HLA-E axis. In this study, we identify HER2-derived peptides that preferentially engage aNK cell responses through the NKG2C-HLA-E axis. Methods Top-scoring peptides were identified from HER2 using neural network—based HLA-E binding models trained on HLA-E variants and NKG2C/NKG2A-specific peptides and compared to reference HLA-E stabilizing peptides using root mean square deviation. Selected peptides were evaluated for in vitro HLA-E stabilization in K562-HLA-E*01:01 cells. NK cell responses from healthy CD3-depleted peripheral blood mononuclear cells (PBMCs) were assessed by intracellular cytokine staining (ICS) following 6-hour peptide stimulation. Results In-silico screening identified three HER2-derived nonamer peptides: PQL, SFVQ, and LLD, with the potential to stabilize HLA-E and preferentially engage NKG2C. In vitro stabilization assays confirmed robust peptide-dependent surface expression of HLA-E compared to DMSO on K562-HLA-E*01:01 cells after peptide stimulation. ICS assay revealed increased CD107a expression on NKG2C+ NK cells compared to NKG2A+ NK cells post peptide-stimulation, indicating NKG2C-mediated cytotoxic responses against HER2 peptides. Conclusion Our findings indicate that HLA-E stabilizing HER2-derived peptides can specifically and robustly activate NKG2C-mediated cytotoxic NK responses in healthy donors. Additional clinical evaluations will be required, but these findings suggest NKG2C-mediated immunotherapeutics may represent attractive personalized therapies for HER2+ breast and other cancers. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Epigenetic mechanisms controlling longevity of T cell immunity 2259637

Abstract Introduction Long-lived T cell immunity can protect against endemic pathogens that result in iterative challenge to the host, but how such repeated activation of T cells impacts on the molecular mechanisms governing memory potential remains largely unclear. Here, we assessed the role of de novo epigenetic programs in shaping the repertoire and recall response of memory T cells in a setting of iterative acute viral infection. Methods Using our previously established CRISPR system for deleting epigenetic regulators (Kang et al, 2024, Science), we optimized an iterative acute viral infection model with LCMV to serially activate Dnmt3a deficient memory T cells. CRISPR-edited Dnmt3a-deficient P14 CD8 T cells were adoptively transferred cells into B6 mice and subjected to acute infections. Based on our previously established acute viral infection, we evaluated the impact of repetitive antigen exposure on memory CD8 T cells differentiation, longevity, proliferation, cytotoxicity and anti-tumor activity. Results Dnmt3a KO retain potentially memory CD8 T cells stronger proliferative capacity during acutely prolonged stimulation, but non-target sgRNA group P14 ratio gradually decrease. Additionally, Dnmt3a knockout T cells displayed sustained memory precursor features, including higher CXCR3, Ly108, and TCF1, with lower PD-1 and Tim-3 expression. Functionally, they produced more IL-2, suggesting enhanced viral clearance and overall T cell survival. These results indicate that loss of Dnmt3a preserves stem-like properties and enhance the long-term persistence of memory CD8 T cells under repetitive antigen exposure, providing an epigenetic mechanism to improve immune durability. Conclusion Our findings show that Dnmt3a KO T cells have increased T cell stemness and durability and indicate that Dnmt3a drives CD8 T cell memory capacity during iterative acute stimulation. These results clarify our research direction that de novo epigenetic programming continues to shape T cell repertoire and recall memory response. Funding Source National Institutes of Health Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)

Functional Profiling of Rheumatoid Arthritis Reveals Disease-Specific Immunomodulatory Responses and Novel Therapeutic Targets 2256858

Abstract Introduction Immune-mediated inflammatory diseases (IMIDs) represent a high unmet clinical need. Despite numerous single cell studies describing IMIDs immune dysregulation, actionable data for drug discovery-relevant novel targets and biology prediction is still limited, which is at least partially due to their static nature. Here, we present a system level approach to reveal IMID-driving biology through functional perturbation modeling of patient-derived immune cells, combined with multi-omics integration. By systematically mapping perturbation responses to underlying molecular networks in realistic disease contexts, our lab-in-the-loop approach enables rapid prediction and experimental validation to uncover mechanism-of-action insights and therapeutically tractable targets in rheumatoid arthritis (RA). Methods As an ex vivo model for the RA synovial niche, we exposed RA and healthy human PBMCs to synovial fluid (SF) from patients with RA, osteoarthritis (OA), or healthy controls. We applied multiplexed high-content imaging to quantify cell-type morphologies, spatial interactions, and activation markers in response to SF exposure alone or under perturbations. By combining functional data with known drug-target relationships and mapping them to protein-protein interactions, we iteratively selected optimal perturbations for validation screens, sequentially uncovering novel disease specific biological insights. Results SF exposure induced RA specific immune cell activation and spatial interactions effects, which were reverted by standard of care. Molecular networks driving cellular responses to RA vs OA SF were integrated using proteomics and single-cell sequencing. Iterative prediction / validation cycles enriched for SF-induced immune inflammation modulators, revealing how functional perturbation mapping can comprehensively profile disease-driving immune networks and identify novel targets. Conclusion We present a novel, generalizable systems-level framework for therapeutic discovery across IMIDs. Funding Source Österreichische Forschungsförderungsgesellschaft (FFG), Austria Wirtschaftsservice (AWS) Topic Categories Computational and Systems Immunology (COMP)

Complement Activation and Anti-Aquaporin-4 Antibodies Drive Pathology in a Preclinical Model of Neuromyelitis Optica Spectrum Disorder 2261227

Abstract Introduction Neuromyelitis optica spectrum disorder (NMOSD) is a severe autoimmune disease of the central nervous system (CNS), characterized by an autoantibody response against the water channel aquaporin-4 (AQP4) on astrocyte endfeet. The disease primarily affects the spinal cord and optic nerves, leading to paralysis and loss of sight. Onset is associated with a breakdown of immune tolerance to AQP4 and the appearance of anti-AQP4 IgG in serum. These antibodies, along with complement deposition, are consistently found in CNS lesions and contribute to astrocyte damage, demyelination, and neurological deficits. The clinical efficacy of plasma exchange and anti-C5 therapies supports the pathogenic role of antibodies and complement in NMOSD. Methods We examined the contribution of anti-AQP4 antibodies and complement in a robust AQP4201-220-induced animal model. Wild-type (WT) and secretory immunoglobulin-deficient (AID⁻/⁻μS⁻/⁻) mice were immunized with AQP4201-220 and treated with pertussis toxin on days −2 and 0. Suboptimal anti-IFN-γ was used to enhance disease; IFN-γ receptor knockout (IFNGRKO) mice were similarly immunized. On day 21, spinal cord and optic nerve were collected for histology, while flow cytometry and transcriptomics assessed immune activation. Results WT + anti-IFN-γ mice showed robust IgG and C5/C5b-9 deposition with CNS upregulation of complement genes (contrasting downregulation in spleen). AID⁻/⁻μS⁻/⁻ mice failed to develop disease, confirming antibody dependence. C3⁻/⁻ mice showed reduced incidence, milder clinical scores, and decreased C5/C5b-9, indicating C3 is required for C5 activation. Anti-C5 treatment delayed onset, reduced severity, and modulated immune responses similarly to genetic C3 loss. Conclusion Our data establish anti-AQP4 antibodies and the C3-C5 (MAC) complement axis as central drivers of NMOSD pathology, reinforcing their value as biomarkers and therapeutic targets. Funding Source R01 AI155678 Topic Categories Neuroimmunology (NEUR)

Endobiotic glycolipid structure of gut symbionts modulates host inflammatory response 2251902

Abstract Introduction Commensal microbes in the intestine continuously shape the immunological landscape by producing diverse microbial products with immunomodulatory potential. Among these, outer membrane glycolipids serve as key mediators of host—microbe interaction. Lipidomic analysis revealed that symbiotic microbes possess diverse lipid A species differing in acyl chain number and phosphorylation degree, whereas pathobionts show limited diversity, predominantly producing hexa-acylated, diphosphorylated species. This compositional complexity across species has hindered detailed investigation of structure—activity relationships. Methods To dissect the immunological roles of symbiotic lipid A structures, we employed chemically synthesized analogs resembling glycolipids from commensals. Results These analogs, varying in acylation and phosphorylation, induced distinct transcriptional responses in dendritic cells. Notably, under-acylated (tri- and tetra-acylated) and monophosphorylated lipid A analogs strongly activated type I interferons and IFN-stimulated genes, whereas penta-acylated species lacked this activity. This structure-driven response correlated with preferential engagement of endosomal TLR4, leading to sustained IFN-β production and intracellular lipid droplet formation. The lipid A—induced IFN-β response was essential for promoting colonic RORγt+ regulatory T cells while suppressing Th17 differentiation and gut inflammation. Consistently, lipooligosaccharides from a B. fragilis strain engineered to overproduce tetra-acylated lipid A showed stronger IFN-β—inducing capacity than those from the wild type. Conclusion Collectively, our work delineates the mechanistic basis linking commensal glycolipid structure to host immunomodulatory responses. Structural nuances in symbiont-derived lipid A variants regulate the maintenance of colonic Tregs, sustaining a balanced and healthy endobiotic state. Funding Source This work was supported by Research Fellows Award from Crohn’s and Colitis Foundation (649279 to Hyoung-Soo Cho), Department of Defense (W81XWH1910625 and HT94252310226 to Dennis L. Kasper.; W81XWH1910626 to Sungwhan F. Oh), and National Institute of Heal Topic Categories Mucosal and Regional Immunology (MUC)

The Role Of B Cell Specific MHC II Antigen Presentation In Breast Cancer Progression And Immune Checkpoint Blockade Therapy 2310330

Abstract Introduction Breast cancer remains the leading cause of cancer-related mortality in women. Immunotherapies have revolutionized treatment; however, their efficacy relies heavily on immune cell-mediated antigen (ag) presentation within the tumor microenvironment (TME). The often overlooked role of B cell ag presentation may be important in anti-tumor immunity. B cells are potent ag presenting cells and therole of B cell ag presentation in progression of mouse models of autoimmune diseases has been demonstrated. B cells may have a potent role in breaking immune tolerance towards self-peptides. We hypothesize that B cell specific MHC II ag presentation plays a non-redundant role in promoting anti-tumor immunity. Methods To investigate the role of B cell MHC II ag presentation in tumor progression, we generated B cell-specific MHC II knockout mice and utilized the E0771, a murine triple-negative breast cancer (TNBC), and E0771-LN26 cell line with a propensity for lymph node and metastases. Tumor growth was assessed in B cell MHC II-deficient and wild-type littermates. We also assessed tumor growth in anti-PD-1 treated condition. To understand the relevance of this study in human TNBCs, we generated co-relation of MHC II expression in breast cancer on patient outcome, we analyzed publicly available TNBC data. Results Our data demonstrate that E0771 and E0771-LN2 tumors exhibit accelerated growth in B cell-specific MHC II knockout mice compared to MHC II fl/fl controls, highlighting a critical role for B cell-mediated MHC II ag presentation in controlling tumor progression. Moreover, T cells from mice lacking B cell specific MHC II expression had were more exhausted. Intrestingly, mice lacking B cell MHC II expression were more responsive to anti-PD-1 treatment. We also found a co-relation of high MHC II expression with better patient outcome. Conclusion These findings suggest that B cell-mediated antigen presentation plays a context dependent role in modulating anti-tumor immune responses in breast cancer. Funding Source This research project is supported by funding from NCI Topic Categories Classical and Non-Classical Antigen Presenting Cells (APC)

Homogenizing Sn–Pb Distribution Through A‐Site MA Cations for Efficient All‐Perovskite Tandem Solar Cells

ABSTRACT Mixed tin–lead (Sn–Pb) perovskites are essential for high‐efficiency tandem solar cells, typically employing a formamidinium (FA)‐dominated composition with a small amount of MA. However, the regulatory role of such A‐site mixing on mixed Sn–Pb perovskite properties remains underexplored. This work demonstrates that engineering the thermodynamic landscape of mixed Sn–Pb perovskite formation enables simultaneous regulation of phase homogeneity and defect chemistry. By controllably incorporating methylammonium (MA) cations, the crystallization kinetics are modulated to promote uniform nucleation, suppress Sn–Pb phase segregation, and inhibit Sn 2+ oxidation, thereby mitigating non‐radiative recombination losses. As a result, mix tin–lead perovskite solar cells achieved a certified power conversion efficiency (PCE) of 23.90%. By leveraging the improved compositional homogeneity and suppressed defect density, the optimized tin–lead absorber is further integrated into two‐terminal monolithic all‐perovskite tandem devices, delivering a PCE of 30.13% (certified 29.57%). The unencapsulated tandem devices maintained 90% of its initial PCE after 495 h of maximum power point operation under simulated one‐sun illumination.

Inherent Microstructural Engineering: Tailoring Amorphous‐Nanocrystalline Composite Films via Ion Beam Technology for Long‐Cycle Lithium Metal Batteries

ABSTRACT Lithium metal anode faces formidable challenges from uncontrollable dendrite growth and unstable solid‐electrolyte interphase (SEI). Interface engineering of the current collectors (CCs) or lithium anodes presents a viable solution. We propose engineering the intrinsic microstructure of coatings to precisely construct interlayers that are both lithiophilic and possess rapid kinetics. A strategy using ion beam deposition (IBD) technology to craft ZnMgSn films with composite microstructures on commercial Cu CCs and lithium foils is reported. This artificial interphase not only exhibits a strongly lithium adsorption energy, but also significantly reduces the diffusion barrier for lithium atoms, thereby synergistically enabling uniform lithium plating. Crucially, this interphase promotes the in‐situ formation of a mechanically robust, bilayer SEI rich in LiF, which can effectively accommodate volume changes during cycling. As a result, the ZnMgSn@Cu CC symmetric cell achieves an ultralong lifespan of over 11000 h. The full cell shows a capacity retention of 85.13% after 130 cycles at 5C. The modified lithium anode maintains over 80% capacity after 620 cycles at 1C. This work not only provides an efficient modification strategy but also offers profound insights into the microscopic design principles for an ideal lithium metal interphase, paving the way for practical lithium metal batteries.

TNAP-mediated hydrolysis of phosphorylated enamel matrix proteins drives local phosphate generation and enamel mineralization

Effects of early adulthood chronic sleep disruption on late life cognition, brain pathology, and microglial reactivity in the PS19 mouse model of tauopathy

Abstract A bidirectional relationship has been proposed between sleep and Alzheimer’s disease pathology, such that disruption to sleep can lead to increases in the expression of tau or its phosphorylation. Over the long term, this can potentially lead to greater expression of neurofibrillary tangles and neurodegeneration. Neurodegenerative diseases characterized by tauopathy can often manifest with deficits in motor and spatial learning. We hypothesized that early adulthood chronic sleep disruption could accelerate tau pathology in a way that alters the trajectory of spatial and motor learning in later life. To test this hypothesis, we subjected early adulthood PS19 mice (MAPT P301S) and wildtype (WT) littermates to automated daily chronic sleep disruption (SD) vs. ad libitum sleep from 2 months to 4 months of age. A 4-day Barnes maze spatial learning task and 2-day rotarod motor learning task were conducted at 6, 8 and 10 months of age with subsequent brain neuropathological evaluation at 10–14 months prior to death. Contrary to our expectations, PS19 mice experiencing early adulthood SD displayed significantly improved spatial learning in comparison to ad libitum sleeping PS19 mice at 8 and 10 months of age, with no significant impact of early adulthood SD in WT mice. At 6 months, there was a significant impairment in offline motor change in PS19 mice compared to WT mice but without a significant effect of prior sleep condition. Early adulthood SD in PS19 mice led to increased hippocampal microglia density, without increased phagocytic microglia density, in late life compared to ad libitum sleep, suggesting sustained microglial proliferation by prior chronic SD. Similarly, TUNEL positive cells were also increased with early adulthood SD in PS19 mice, suggesting persistent DNA damage in late life. However, neither tau pathology nor histological markers for neurodegeneration were significantly different between early adulthood sleep conditions in PS19 mice tested at later life. Thus, the improvement in spatial learning in late life following early adulthood chronic SD in PS19 mice would seem disentangled from an effect on tauopathy or neurodegeneration, but whether the behavioral consequence is related to the observed changes in hippocampal microglial density remains to be mechanistically linked.

CXCL10 orchestrates immunopathogenic CD4+ effector recruitment in alphavirus arthritis 2259213

Abstract Introduction O’nyong-nyong virus (ONNV) is a mosquito-transmitted alphavirus closely related to chikungunya virus (CHIKV) that causes fever and debilitating polyarthritis in humans. To dissect immunopathogenic mechanisms driving ONNV-induced disease, we modeled infection in C57BL/6 mice. During the subacute phase (10 days post-infection), persistent viral RNA in the footpad coincided with a CXCL10-dominated chemokine milieu and pronounced infiltration of CD4+ T cells displaying an effector (CD44+CD62L-) phenotype, whereas CD8+ T cell recruitment was restricted. We sought to further explore the role of CXCL10 in this early phase of the disease. Methods We used C57Bl/6 mice to model ONNV infection, inoculating mice with 10^5 PFU of the virus through footpad injections. To perturb CXCL10 signaling we intraparitoneally administered an anti-cxcl10 antibody or atorvastatin. We collected infected footpads, draining lymph nodes and spleens at 10 days post infection to profile cellular composition and phenotype via flow cytometry and the expression of inflammatory markers by RT-qPCR. Results In the draining popliteal lymph node, CD4+ but not CD8+ T cells upregulated CXCR3 and activation markers, consistent with selective chemotactic responsiveness to CXCL10. Functional perturbation of this axis, using a CXCL10-neutralizing antibody or pharmacologic downregulation via atorvastatin, significantly reduced CD4+ and monocyte accumulation within the infected footpad, while enhancing activation and functional markers on lymph node CD8+ T cells. Atorvastatin treatment also decreased local viral burden, implicating CXCL10-driven inflammation in sustaining viral persistence Conclusion These findings identify the CXCL10-CXCR3 signaling axis as a key immunoregulatory node that skews the local immune response toward pathogenic CD4-driven inflammation at the expense of cytotoxic antiviral immunity. Modulating this chemokine circuit may recalibrate local immunity and mitigate alphavirus-associated arthritic disease. Funding Source n/a Topic Categories Viral Immunology (VIR)

Preliminary immunologic and clinical Phase 1b study outcomes from patients with ROSAH syndrome treated with the first-in-class ALPK1 inhibitor DF-003 2307448

Abstract Introduction ALPK1 (alpha-protein kinase 1) is a key gene and protein involved in innate immunity and inflammation. ROSAH syndrome is an autosomal dominant disorder caused by pathogenic variants in ALPK1 (e.g., T237M) and characterized by ocular inflammation, headache, anhidrosis, splenomegaly, and systemic manifestations. Elevated levels of hsCRP and inflammatory cytokines, including CCL2, CXCL10, TNFα, IL-6, IL-1β, and IL-8, have been observed in affected patients. DF-003 is a first-in-class oral ALPK1 inhibitor under development for treating the genetic root cause of ROSAH syndrome. Methods This Phase 1b trial (NCT06395285) is a multicenter, single-arm, open-label study evaluating the safety and tolerability of DF-003. Six patients with ROSAH syndrome (T237M ALPK1) aged 18—65 received daily oral DF-003 for 4 weeks. Serum inflammatory biomarkers and clinical outcomes including quality of life (QoL), assessment of anhidrosis and arthralgia were evaluated. Spleen changes were evaluated by CT/MRI with the Mint Lesion™ system. Results Four out of 6 subjects reported the reversal of anhidrosis, improvements in QoL (reduced EQ-5D scores), and improvement in arthralgia. These benefits diminished following discontinuation of therapy. Of the 2 subjects with available immunological marker data, 1 whose markers were elevated at baseline showed pharmacodynamic improvement, with reductions in IL-6, IL-8, CXCL-10, and hsCRP to within normal ranges after 28 days. In the 1 subject that did not have splenectomy, there was a ∼20% reduction in spleen volume that reversed upon cessation of drug. All laboratory values remained within normal limits, with stable renal and hepatic function, and no serious or treatment-emergent adverse events were reported. Pharmacokinetic data available from 6 subjects support once-daily dosing. Conclusion Inhibition of ALPK1 with DF-003 demonstrated early evidence of clinical and pharmacodynamic improvements without safety concerns. These results warrant evaluation of DF-003 in a Phase 2 trial. Funding Source All funding was provided by Drug Farm Topic Categories Translational and Interventional Immunology (TI)

Engineered probiotics limit T-cell driven CNS autoimmunity by activating AHR-KLF4-FADS3 signaling in intestinal dendritic cells 2235071

Abstract Introduction Dendritic cells (DCs) control effector immune responses and tolerance. Their function is regulated by transcriptional, epigenetic and metabolic programs, but the mechanisms controlling DC metabolism and its impact on autoimmune diseases remain poorly defined. Methods We used single-cell and bulk RNA-sequencing of multiple sclerosis patients and preclinical models in combination with in vivo DC-specific gene perturbations and in vitro human systems. Results We identified an aryl hydrocarbon receptor (AHR) - Krüppel-like factor 4 (KLF4) - fatty acid desaturase enzyme 3 (FADS3) axis that drives docosahexaenoic acid (DHA) production in DCs, limiting pathogenic T-cell responses and experimental autoimmune encephalomyelitis (EAE) development. Specifically, AHR drives KLF4 expression in conventional DCs (cDCs); AHR and KLF4 cooperate to induce FADS3 expression, promoting DHA synthesis. AHR, KLF4 or FADS3 inactivation in cDCs decreases DHA production, resulting in increased encephalitogenic T-cell responses and EAE worsening. To evaluate therapeutic potential, we engineered a probiotic (EcNIAA) that produces the AHR agonist indole-3-acetic acid (IAA). Oral EcNIAA activates AHR-KLF4-FADS3 signaling in intestinal cDCs, boosted DHA production, and suppressed encephalitogenic T-cells migrating from the gut to the central nervous system. Conclusion Briefly, we identified a novel immunometabolic axis in DCs to limit effector T-cell responses and can be targeted with engineered probiotics to treat autoimmunity. Funding Source National Multiple Sclerosis Society (FG-2307-42209) and Mayer Foundation Topic Categories Neuroimmunology (NEUR)