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IFNγ-Driven Th17 and Neutrophil Responses Promote Rapid Polycystic Kidney Disease Progression 2304963
Abstract Introduction Rapidly progressing polycystic kidney disease (PKD) leads to earlier loss of kidney function and an increased likelihood of kidney transplantation, however the mechanisms driving rapid disease progression are poorly understood. Single-cell RNA sequencing data from our laboratory identified interferon gamma (IFNγ) as a potential mediator of accelerated cyst growth following kidney injury. Based on these findings, we hypothesized that IFNγ promotes rapidly progressing PKD by driving a pathogenic inflammatory immune response. Methods We tested our hypothesis in two PKD mouse models. We first crossed our PKD mouse model to IFNγ-deficient mice, followed by inducing folic acid (FA) kidney injury, and disease progression assessment 56 days post-FA. The second model utilized a non-injured rapidly progressing PKD model which received anti-IFNγ antibody or isotype control 3 times weekly for 3 weeks, before kidneys were collected and disease progression assessed. Results Genetic deletion of IFNγ significantly attenuated injury-accelerated PKD severity, as demonstrated by reduced kidney weight to bodyweight ratio and cystic index. IFNγ deficiency was also associated with decreased kidney fibrosis and improved kidney function. Flow cytometric analysis revealed reduced proportions of neutrophils and Th17 cells, while qPCR demonstrated significantly lower expression of Th17- and neutrophil-associated cytokines. Consistently, antibody-mediated blockade of IFNγ in the non-injured Pkd2f/f model led to a significantly reduced cyst burden and immune cell infiltration, mirroring the genetic knockout findings Conclusion These findings suggest that IFNγ is a key mediator of rapid PKD progression. IFNγ-driven Th17 and neutrophil responses may represent a critical immunopathogenic axis underlying aggressive disease, highlighting this pathway as a potential therapeutic target for slowing disease progression, preserving kidney function, and delaying the need for kidney transplantation. Funding Source N/A Topic Categories Immune Mechanisms of Human Disease (HUM)
Novel agonist monoclonal antibody targeting GPCR to decrease neutrophil accumulation and initiate resolution of chronic inflammation 2310040
Abstract Introduction Resolution of inflammation is an active process mediated by specialized receptors such GPCRs expressed on neutrophils, monocytes, and macrophages. Natural and synthetic GPCRs agonists demonstrate significant preclinical efficacy but face limitations in stability, synthesis, and receptor selectivity. Methods New antibodies were generated and characterized for high affinity and GPCR specificity. Agonist signaling and activity was assessed on cell lines and primary neutrophils and monocytes, and efficacy was evaluated in multiple clinically relevant murine models of acute and chronic inflammation. Results We developed first-in-class antibodies acting as a selective GPCR agonist for the treatment of acute and chronic inflammatory diseases with stable properties and selectivity. This therapy allows a specific agonistic signal and promotes the hallmark pro-resolving activity by limiting activation and recruitment of neutrophils and monocytes/macrophages in contrast to natural or synthetic lipids that can trigger both pro- and anti-inflammatory signals in multiple GPCRs. In an acute lung injury model, the mAb markedly inhibits neutrophils and inflammatory monocytes infiltration into inflamed tissue. In chronic models, treatment alleviates clinical severity, confirming robust therapeutic potential. Conclusion This first-in-class GPCR specific agonist mAb introduces a novel strategy to actively drive long term resolution of inflammation. Its receptor specificity, strict agonist activity and robust preclinical efficacy on neutrophils and inflammatory monocytes recruitment position it as a promising therapeutic candidate for treating acute and chronic inflammatory disorders. Funding Source n/a Topic Categories Cellular Adhesion, Migration, and Inflammation (CAM)
Single cell characterization of early innate immune responses induced by acute and chronic viral infections 2267604
Abstract Introduction Lymph-borne viruses penetrate the host through the skin or mucosa barriers and drain to the nearest lymph node. The innate immune response induced at the infection site and draining lymph node curb virus systemic dissemination and activate adaptive lymphocytes. However, acute and chronic infections caused, respectively, by the mouse pathogens poxvirus ectromelia virus (ECTV) and the murine cytomegalovirus (MCMV) induce very dissimilar immune responses. Thus, we compared the innate immune responses induced at early stages of infection with these two DNA viruses in the draining lymph nodes of wild-type C57BL/6 inbred strain infected with ECTV or MCMV by footpad inoculation. Methods For that, we used We used unbiased cellular indexing of transcriptome and epitope single cell sequencing (CITEseq) to characterized changes induced by infection and infected populations. Results We found that ECTV induced expansion and activation of Natural Killer (NK) cell populations and inflammatory monocytes while MCMV did not. We also observed the presence of a distinct infected cluster in dLN of mice infected ECTV while MCMV was dispersed throughout multiple cell populations that presented low MCMV transcripts. Conclusion Overall, we dissected that acute infection with ECTV induces innate immune responses that correlate with survival while infection with MCMV induced poor innate immune responses. Funding Source n/a Topic Categories Viral Immunology (VIR)
Investigation of immune mechanisms driving the pathogenesis of chronic chikungunya virus disease 2254154
Abstract Introduction Arthritogenic alphaviruses such as chikungunya virus (CHIKV) cause chronic inflammatory arthritis affecting thousands globally, yet there are no effective targeted therapies. Persistent joint inflammation despite viral clearance suggests a critical role for virus-specific, immune-mediated mechanisms. However, immune mechanisms underlying the development of chronic arthritic disease post chikungunya infection is currently unknown. Recent work suggests a pathogenic role for TNF+-producing CHIKV-specific CD4+ T cells in chronic disease and macrophages as the primary target of CHIKV infection. Therefore, it is essential to uncover the molecular crosstalk between different immune cell subsets and define their contribution to disease pathogenesis in humans. Methods Using high resolution spectral flow cytometry and in vitro co-cultures, we dissected the phenotype and function of diverse immune cell populations, including monocytes, dendritic cells, NK cells and T cells in PBMCs from CHIKV-infected individuals with chronic chikungunya disease compared to those who recovered post-infection. Results We identified the functional profile of circulating monocytes, dendritic cells and NK cells in individuals with chronic chikungunya disease and identify the key cytokines and molecules that are released by these immune cell subsets as a result of T cell activation. Additionally, we observe a high production of TNF+, IL-10 and GM-CSF and a lower expression of HLA-DR in monocytes, indicating a pro-inflammatory profile that is exclusive to individuals with chronic disease. This will allow us to understand how monocyte activation affects T cell differentiation and function. Conclusion Here, we identify the key cytokines and molecular players that mediate immune crosstalk and allow differentiation of CD4+ T cells into pro-inflammatory cells. This work will uncover immune mechanisms that drive alphavirus-mediated chronic arthritic diseases. Funding Source n/a Topic Categories Viral Immunology (VIR)
Investigating Functional Mechanisms of Noncoding Genetic Variations in the Human Fc Gamma Receptor Locus 2260728
Abstract Introduction Genetic variations in the low-affinity Fc Gamma Receptor (FcγR) region have been associated with various diseases and treatment efficacies. Using a high-throughput in vitro screen called Regulatory-Element Sequencing (Reel-Seq) we have identified over 400 candidate functional Single Nucleotide Polymorphisms (fSNPs) from the genomic region. In this study, we aim to identify regulatory function of these candidate non-coding fSNPs in THP-1 monocytes. Methods At each candidate FcγR fSNP position, we introduced random mutation in the THP-1 monocyte line using CRISPR-Cas9 based genome editing and generated over 30 knockout clones per fSNP site. We stimulated these THP-1 knockout clones with various treatments including Interferon and Lipopolysaccharide (LPS). Flow cytometry and qPCR were performed to measure the changes in cell-surface FcγR protein abundances and FcγR transcript abundances. Antibody-Dependent Cellular Phagocytosis (ADCP) assays are being conducted on selected clones to evaluate whether normal phagocytosis function has changed through the alteration of FcγR abundances. Results We have observed significant changes in cell-surface FcγR abundance in these THP-1 knockout cells based on flow cytometry analysis. Detailed analysis of stimulated cells is ongoing, and we will share the full results at the meeting. Conclusion Noncoding genetic variants identified by our in vitro screen may play important roles in monocyte function, through the regulation of cell-surface FcγR abundance. Funding Source NIH NIAID #1R01AI187318-01, NCSU CVM Internal Grant 2022, CMI-EID Associate member Professional Development Scholarship 2023 Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)
Machine Learning‐Guided High‐Efficiency and Thermally Stable Capacitive Energy Storage in Dielectric Capacitors With a Simple Chemical Composition
ABSTRACT Dielectric capacitors offering ultrafast charge–discharge capability and superior reliability are essential for advanced electronic systems, but achieving both high energy density and efficiency within simple and eco‐friendly compositions remains a great challenge. Here, guided by machine learning, we achieve high‐efficiency and thermally stable capacitive energy storage in strontium titanate‐based ceramics. Through synergistic local structural engineering and optimized fabrication process, the materials exhibit enhanced polarization, reduced hysteresis loss, and improved breakdown strength. The designed materials deliver an ultrahigh energy density of 10.69 J cm −3 with a near‐ideal efficiency of ∼97% and a record high figure of merit of 392 J cm −3 at room temperature, and retains high performance at 150°C with a figure of merit of 152 J cm −3 and an efficiency of ∼94%. This remarkable performance arises from the incorporation of Bi 3+ ions with high polarizability at the A‐sites of strontium titanate quantum paraelectrics, which breaks structural symmetry and induces lattice and octahedral distortions, leading to the formation of nanoscale polar clusters with highly dynamic fluctuations. These findings establish a compositionally simple, environmentally benign pathway for developing dielectrics with superior energy storage capability and thermal stability, offering new opportunities for high‐performance capacitive energy storage systems.
Observing positive and negative social touch triggers topographically distinct physical and emotional reactions in the body
Abstract Interpersonal touch is a fundamentally multidimensional and multisensory experience and as such is highly subjective. By whom, where and how we are touched determines how we react to and feel about it. Previous research using topographical body tasks has mainly examined where people feel comfortable being touched by others. Using the BodyMap Tool, we mapped distinct somatoperceptions associated with positive and negative social touch videos, comparing physical sensations and emotional responses. We quantified these patterns with a BodyMap Score (BMS) reflecting valence and spatial coverage. Positive videos elicited higher physical than emotional BMS, whereas negative videos showed the opposite pattern, with broader emotional than physical somatoperceptions. Individual differences further modulated these effects: touch-avoidant participants showed reduced BMS for positive touch, and participants reporting pain exhibited overall lower BMS, though this reduction was attenuated for positive stimuli. These findings highlight the complex and context-dependent bodily representation of social touch.
Stability and Progressive Differentiation of Tfr cells are intrinsically and extrinsically controlled by Tfh programs 2255794
Abstract Introduction Follicular regulatory T (Tfr) cells restrain follicular helper T (Tfh) cell-mediated B cell responses in the germinal center (GC) reaction to optimize humoral immunity while limiting autoimmunity. The immune system partially regulates GC responses by controlling the stepwise differentiation of Tfh cells. Whether Tfr cell development requires sequential developmental stages and how the immune system regulates these processes to control humoral immunity is poorly understood. Methods We used longitudinal sampling of lymphoid organs along with fate mapping and matched single-cell RNASeq/TCRseq to assess developmental dynamics of Tfr cells. Results We found that Tfr cells undergo dynamic clonal expansion and Tfr cells also undergo progressive differentiation through progenitor-like, early effector, and late effector stages. Late effector Tfr cells possess inherent instability, with a propensity to lose expression of the transcription factor FoxP3 to become ExTfr cells. ExTfr have unique features and can be redeemed to become suppressive Tfr cells. Acquisition of a Tfh-like transcriptional program in Tfr cells was an intrinsic predictor of progeny instability. Extrinsically, Tfh cells enhanced late effector Tfr cell differentiation by diverting early effector Tfr cells away from a default Blimp1-expressing pathway to a Bcl6-expressing one. Tfr cells are a dynamic and plastic cell subset, the progressive differentiation of which is controlled at later effector stages by intrinsic and extrinsic programs that work together to provide a negative feedback loop to control humoral immunity. Moreover, Tfh induce Tcf7 in Tfr cells which promotes late effector Tfr differentiation Conclusion Tfr cells are a dynamic and plastic cell subset, the progressive differentiation of which is controlled at later effector stages by intrinsic and extrinsic programs that work together to provide a negative feedback loop to control humoral immunity. Funding Source n/a Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)
Inflammasome formation in bile duct from rat clonorchiasis 2334111
Abstract Introduction Clonorchis sinensis is the liver fluke to infect human liver bile duct for clonorchiasis and persist for more than 27 years. The worms induce severe inflammatory reaction on bile duct and cause the one of accelerating factors for cholangiocarcinoma. As zoonotics, that worms infect and induce rat clonorchiasis. Inflammasome is the immune complex to induce inflammation with cytokines of IL-1beta, IL-18 and TNF alpha and several others. Methods To comprehend the inflammation on bile duct induced by C. sinensis habit, the cytokine factors for inflammasome was analyzed with bile duct infiltrated by inflammatory cells, using real time RT-PCR. The thick inflammatory bile duct tissues from three points including common bile duct and hepatic bile ducts were harvested and applied to the process of RNA extraction. For a comparison, the small intestine tissue of one point was selected. Rat GAPD was used as a house keeping gene. The real time PCR for 40 cycles. Results The results showed that three samples of bile ducts showed different levels of IL-1 beta and lower than that of intestine. Three samples of bile ducts showed much similar in TNF-alpha and much lower than that of intestine. Two samples of bile ducts showed different levels of IL-18 and lower than that of intestine. Conclusion These results indicated that on inflammatory rat bile ducts induced rat clonorchiasis, the levels of TNF alpha are similar among three bile duct points and the level of IL-1 beta and IL-18 are much different depending on points. Funding Source N/A Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)
Novel KIR:HLA interaction (KIR2DS4:HLA-B*35) predicts patient survival post haploidentical donor transplantation 2252772
Abstract Introduction Haploidentical hematopoietic cell transplantation(haploHCT) has become a mainstay in our clinical care for patients with leukemia. Upon the discovery of activating and inhibitory killer immunoglobulin receptors(KIRs) on NK cells and their cognate human leukocyte antigen(HLA) ligands, algorithms have been developed to enhance a graft versus leukemia effect. However, these do not yield consistent predictions in patient outcomes. We posit that this is due to an incomplete map of the entire KIR:HLA interactome. Methods A combination of in silico protein folding and interactions to determine KIR:HLA reactivity in addition to acoustic force microscopy measuring cell avidity(CA) as a readout for KIR signal strength were used. Cell avidity was determined using monoallelic HLA expressing K562 cell lines along with KIR typed ex vivo expanded peripheral blood NK cells. We then applied the newly discovered interaction to patients who had undergone a single haploHCT at our institution with or without additional NK cell infusions post-transplant. Results We found that KIR2DS4 and HLA-B*35 interact in silico. We confirmed this through CA measurements. We took this interaction and sought to determine if it was clinically significant. When stratifying pediatric and adolescent young adult patients based on their HLA B*35 positivity and the donor KIR2DS4 expression, we delineated a strong correlation to survival (P = 0.061) when donors had two copies KIR2DS4. When patients received haploHCT and additional NK cell addback from donors with two copies of KIR2DS4 they did not die or experience relapse during follow-up, resulting in longer OS(P = 0.016) and RFS(P = 0.001) than patients with donors with one copy KIR2DS4. Conclusion We describe a novel KIR2DS4:HLA-B*35 interaction axis that predicts patient survival and lends strong credence to revisit the previously defined KIR ligands and their signaling. Our approach is immediately clinically implementable and warrants further exploration of the KIR:HLA interactome. Funding Source Edward P Evans Young Investigator Award Topic Categories Transplantation Immunology (TRAN)
Use of bacterial chassis for the intratumoral delivery of a super-interferon gamma in a breast cancer model 2306491
Abstract Introduction Gamma interferon (IFNγ) is a key cytokine during in inflammation and cancer. In fact, studies showed an increased survival rates in those patients with high levels of tumor-infiltrating IFNγ. However, its high aggregation when produced recombinantly, as well as its short half-life, requires of high administration doses, resulting in toxicity and limiting its clinical use. In our lab, we generated a super-human IFNγ by introducing point mutations which showed an enhance biological activity. Furthermore, we used an attenuated bacterial chassis developed in our lab to locally deliver this IFNγ directly intratumorally in a rodent breast cancer model. Methods Computational tools (FoldX and ModelX) were used to predict mutations with increased stability and receptor binding. All variants were secreted by our chassis, and its activity was tested using HEK-Blue IFNγ reporter cells. Inflammatory and anti-tumor effects were evaluated in THP1 monocytes and MDA231/MCF7 tumor cell lines. In vivo studies were performed in WT and NOD-scid mice after mammary tumor inoculation followed by intratumoral treatment with IFNγ-secreting chassis. Results We generated a human IFNγ mutant with a 10-fold higher activity and 7-fold improved production by our chassis. The variant showed greater thermal stability and stronger inflammatory and anti-proliferative effects in cell lines. In vivo, it significantly increased immune-cell recruitment and boosted inflammatory gene signatures within the tumor environment. Conclusion Our chassis can efficiently secrete functional IFNγ, and computationally predicted mutations resulted in a super-IFNγ with enhanced stability, production, and activity. Super-IFNγ intratumoral delivery resulted in enhanced immune cell infiltration and an increased inflammatory genes expression in vivo. Future work with in vivo models will extend tumor environment effects and currently, we are evaluating the possibility to combine IFNγ with other immunotherapies such as cancer vaccinology. Funding Source ERC grant Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)
NEMO is rapidly recruited to the TCR via polyubiquitin chain recognition and is dissociated via IKKβ kinase activity 2310337
Abstract Introduction The IκB kinase (IKK) complex is best known for its roles in cell survival and in the activation of canonical NFκB isoforms. The complex contains two kinase subunits (IKKα/β) and a dimeric polyubiquitin-binding adaptor subunit known as the NFκB essential modulator protein (NEMO) [IKBKG]. While oversimplified models often place the IKK complex entirely downstream of the adaptor protein Carma1, T cell receptor (TCR)-induced IKKβ phosphorylation is Carma1-independent. Thus, IKK complexes may play unconventional roles downstream of the TCR, but upstream of Carma1. Methods Here, we examine how IKK complexes participate in TCR proximal signals via dynamic imaging analyses of fluorescent NEMO chimeras expressed in Jurkat T cells and primary human T cell blasts. Results We observe that NEMO enters TCR- and ZAP-70-containing microclusters within ∼70 seconds of their formation, via processes that rely on the activities of Lck and ZAP-70, and that are independent of downstream effectors implicated in canonical NFκB activation. The mutational inactivation of either the K63 or the linear polyubiquitin-binding sites in NEMO, the pharmacological inhibition the K63-chain forming E2 ligase Ubc13, or the inhibition of the linear polyubiquitin forming LUBAC complex all prevent the recruitment of NEMO to the TCR. If NEMO is co-expressed with wild-type IKKβ, the co-clustering of NEMO with the TCR is largely eliminated. However, kinase dead IKKβ co-clusters with NEMO at the TCR, and IKKβ inhibitors shift wild-type IKK complexes into TCR microclusters. Conclusion These data suggest that the IKK complex is rapidly recruited to the TCR via multipoint interactions with ubiquitin polymers assembled within ∼1 minute of ZAP-70 recruitment, whereas the activity of the IKK complex releases NEMO from the TCR. These findings define a window within which partially activated IKK complexes may influence TCR-proximal signaling events upstream of Carma1. Funding Source NIH R01 AI103022 Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)
Mouse EL4 lymphoma strains differentially respond to a loss of BCATc from T cells and αCTLA4 immunotherapy 2302854
Abstract Introduction Lymphoma tumors can escape immune surveillance and grow rapidly. One of the features of this plasticity is immunosuppressive signals causing T cell exhaustion. Exhausted T cells upregulate the CTLA4 receptor, which dampens the immune response. Use of αCTLA4 reverses this process, however, αCTLA4 therapy has limited application and is currently not offered to lymphoma patients. This calls for research into deciphering the molecular barriers in lymphoma tumors. The cytosolic branched-chain aminotransferase (BCATc), an enzyme that breaks down leucine, may contribute to immunosuppression within the lymphoma microenvironment. Methods To understand how BCATc influences the efficacy of αCTLA4 therapy, T-BCATcfl/fl mice and mice with a loss of BCATc from T cells (T-BCATcKO) were subcutaneously injected with 2x10^5 of low (EL4) and high (EL4OVA) immunogenic lymphoma and randomly assigned 10 μg/gram body weight i.p. injections of IgG isotype or αCTLA4 on days 2, 5 and 10-post lymphoma challenge. Mice were euthanized on day 15 and T cell populations in tumors and draining lymph nodes were analyzed by flow cytometry. Results Prior to αCTLA4 therapy, T-BCATcKO mice responded to EL4 and EL4OVA lymphoma with a significant 40-50 % delay in tumor development. This was further reduced when T-BCATcKO mice were subjected to αCTLA4 following inoculation with EL4OVA lymphoma. 78 % of these mice showed a 5-fold reduction in tumor masses compared to mice injected with isotope antibody. In contrast, only 12.5 % of T-BCATcKO mice carrying EL4 tumors responded with reduced tumor volumes to αCTLA4; the rest of the mice developed ∼40 % larger tumors with significantly reduced percentage of memory precursor CD4+ T cells compared to T-BCATcKO mice injected with IgG, or T-BCATcfl/fl mice treated with αCTLA4. Conclusion These findings stress the differential response of low and high immunogenic lymphomas to BCATc expression and may have clinical implication in targeting BCATc for combinatorial immune therapies in humans. Funding Source NCI-1R15CA249796-01A1 Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)
Inhibition of Tonic PD-1 Signaling in Dendritic Cells Enhances Costimulatory Pathways 2254769
Abstract Introduction Immune checkpoint blockade therapies, particularly those targeting the PD-1/PD-L1 and CTLA-4 pathways, have fundamentally transformed cancer treatment, achieving clinical responses in approximately 25% of patients. Although combinatorial strategies have improved these outcomes, a substantial proportion of patients remain refractory to treatment or develop resistance. Emerging evidence implicates tonic PD-1 signaling–a ligand-independent, constitutive pathway distinct from canonical checkpoint inhibition–as a potential contributor to therapeutic resistance. To elucidate the impact of tonic PD-1 signaling on dendritic cell (DC) function and to explore innovative therapeutic approaches aimed at restoring their immunogenic capacity. Methods We utilized lentiviral transduction to generate DC2.4 dendritic cell lines stably overexpressing PD-1. Under conditions of tonic PD-1 signaling, we evaluated cell viability and the expression of key costimulatory molecules (MHC II, CD80, CD86). Additionally, we investigated the effects of RIPR-PD-1, a bispecific molecule that disrupts tonic PD-1 signaling via CD45 tethering, and SHP009, a pharmacological inhibitor targeting SHP2 recruitment. Results PD-1 overexpression in DCs led to a significant reduction in both cell viability and the expression of costimulatory markers. Treatment with RIPR-PD-1 or SHP009 effectively reversed these inhibitory effects, restoring DC viability and promoting the upregulation of MHC II, CD80, and CD86 expression. Conclusion These findings demonstrate that tonic PD-1 signaling compromises dendritic cell functionality by attenuating their viability and costimulatory potential. Pharmacological blockade of tonic PD-1 signaling–either through RIPR-PD-1 or SHP2 inhibition–reinstates a pro-immunogenic DC phenotype. Targeting this non-canonical PD-1 pathway represents a promising strategy to potentiate antitumor immune responses and overcome resistance to existing immunotherapies. Funding Source n/a Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)
Absence of the “Don’t Eat Me” signal results in aberrant T cell responses to Leishmania major 2260958
Abstract Introduction CD47 is a ubiquitously expressed cell surface protein that regulates immune homeostasis by engaging signal regulatory protein alpha (SIRPa) on phagocytes to inhibit their activation. While the CD47-SIRPa axis is known to regulate both innate immunity and T cell responses in cancer & sterile inflammation, its role in infectious disease is poorly defined. Leishmania major (L. major) is an intracellular protozoan parasite that infects phagocytes. Macrophages and CD4+ T cells are both critical for immunity to L. major infection, so we interrogated the role that “don’t eat me” signals play in immunity to L. major. Methods We infected Cd47-/- & Sirpa-/- mice on a C57BL/6 background subcutaneously in each footpad and harvested draining lymph nodes & spleens at several critical time points for flow cytometry. T cell responses were assessed further by ICS & transcription factor staining. Results Cd47-/- & Sirpa-/- disease is severe. They present increased lesions, swelling, & parasite burden compared to controls. Decrease in T cells & increase in antigen experienced T cells is seen. IFNg & TNFa producers are reduced, likely liable for the severity. Conversely, T cell activation is increased in KO mice during acute disease (15dpi) when severity is similar. Conclusion Further studies are in progress to characterize why Cd47-/- & Sirpa-/- mice fail to control L. major infection despite strong early Th1 responses. This is a novel finding showing CD47-SIRPa is required for protective T cell responses to infection. Funding Source University of Iowa Mechanisms in Parasitism Predoctoral T32 Fellowship, T32 AI007511 NIH R01AI155425, VAI0BX006091 Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)
Optoelectronic Nanofluidic Neural Networks for Ionic Computing
ABSTRACT Emerging neuromorphic computing technologies aim to replicate the brain's cognitive processes and offer promising capabilities for advanced computation and complex tasks. Building artificial neural networks (ANNs) with robust connections and dynamic plasticity is essential for their implementation. Nevertheless, most existing ANNs rely on solid‐state semiconductors, where electrons or holes serve as information carriers. Here, we present a novel ion‐based nanofluidic memristor operating in aqueous solutions, which can be integrated to create densely connected ionic neural networks (INNs). The nanofluidic memristor exhibits tunable ionic memory and diverse synaptic plasticity that can be modulated by electrical, optical, or combination signals. We demonstrate that the ionic memory originates from non‐trivial hysteretic ion‐charged surface interactions in confined nanochannels, supported by experiments and numerical calculations. Utilizing the tunable states of the nanofluidic memristor, we implement reservoir computing to classify static patterns and dynamical motions, achieving ultrahigh accuracies of 91% and 97% on the standard MNIST datasets and homemade moving‐particle library, respectively. Multiple nanofluidic memristors are integrated to construct a practical INN capable of real‐time logic computation and in‐sensor computing, verifying the robustness of nanofluidic computation systems. This ion‐based nanofluidic platform offers a promising pathway towards fully connected INNs and advances neuromorphic computing beyond solid‐state electronics.
The effect of dynamic motion and simultaneous presentation on the statistical learning of nonadjacent dependencies in manual gesture sequences
Abstract Statistical learning enables humans to extract regularities from sensory input, but its efficiency depends on how information is structured within each modality. In a preregistered study, we tested how temporal/spatial organisation and motion cues shape non-adjacent dependency learning of sequences of sign-like manual gestures. We manipulated presentation type (serial vs. simultaneous) and dynamicity (dynamic vs. static). Participants completed an online target-detection task, in which changes in reaction times and accuracies indexed learning, followed by an offline grammaticality-judgment task. Across conditions, participants ( N = 123) showed evidence of sensitivity to the underlying structure: reaction times decreased during training, increased when structured input was replaced with random sequences, and offline accuracy exceeded chance. Simultaneous presentation produced larger online improvements during exposure, although offline grammaticality judgements did not differ across conditions. Motion cues did not improve model fit. These findings suggest that visual statistical learning for structured manual gesture input benefits more from simultaneously available spatial information than from sequential presentation, suggesting that optimal input structure depends on modality-specific perceptual constraints.
Neutrophil Formyl Peptide Receptor-1 Activation Triggers Endogenous Opioid Release for Local, Non-addictive Analgesia 2260008
Abstract Introduction Pain remains a major clinical challenge; while opioid analgesics are effective, their use is constrained by tolerance, dependence, and respiratory depression. Immune cells both sense injury and secrete analgesic mediators at inflamed sites, enabling local modulation with minimal systemic exposure. Among these mediators, endogenous opioid peptides (e.g., Met-enkephalin) can provide potent peripheral analgesia without central nervous system toxicity. Activation of the Formyl Peptide Receptor-1 (FPR1) on neutrophils by bacterial peptides such as fMLF initiates a Phosphoinositide 3-Kinase (PI3K)-driven signaling cascade that mobilizes intracellular Ca²+, triggering Met-enkephalin release. We hypothesized that activating FPR1 on neutrophils would physiologically trigger Met-enkephalin release, enabling localized, non-addictive analgesia. Methods A library of 140 small-molecule FPR1 agonists was designed and synthesized to identify candidates capable of inducing endogenous opioid release. Compounds were screened in HL60 cells differentiated toward a neutrophil-like phenotype and in primary human neutrophils. Functional responses were quantified by intracellular Ca²+ mobilization (FPR1 signaling readout) and Met-enkephalin release measured by ELISA and LC—MS/MS. Results Lead compounds produced robust, concentration-dependent calcium flux in differentiated HL-60 cells with EC50 values ranging from 1.46—13 µM. Agonist stimulation promoted Met-enkephalin release to ∼100 pg/mL in cell-based assays. Preliminary studies in primary human neutrophils confirm agonist-evoked Ca²+ mobilization, with opioid peptide-release. Conclusion Small-molecule activation of neutrophil FPR1 can elicit endogenous opioid peptide secretion, revealing a neuroimmune pathway for site-specific analgesia. By leveraging innate immune trafficking and secretory programs, this approach may enable peripherally acting, opioid-mediated pain relief while minimizing systemic adverse effects, motivating in vivo efficacy and safety studies. Funding Source NIH Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)
Functional and dysfunctional T regulatory cell states in human tissues in RA and other autoimmune arthritic diseases 2334965
Abstract Introduction Regulatory T cells (Tregs), characterized by FOXP3 expression, are essential for maintaining immune homeostasis by controlling inflammation. However, in autoimmune diseases such as rheumatoid arthritis (RA), impaired Treg function contributes to immune dysregulation and disease pathology. The phenotype of impaired Treg function in RA and other autoimmune diseases remains unknown. Methods We employed unbiased paired single cell RNA-sequencing (scRNAseq) and T cell receptor (TCR)-seq analysis of Tregs from synovial tissue, synovial fluid, and blood from 79 RA patients and blood from 5 healthy donors in an integrated analysis. We mapped Tregs from additional inflammatory arthritic conditions, Juvenile Idiopathic Arthritis (JIA) and Psoriatic Arthritis (PsA), to evaluate the Treg heterogeneity across different conditions. Results We identified two predominant Treg states, CD25hiCXCR6pos Tregs with strong suppressive function, and CD25loAREGpos Tregs, a dysfunctional state exclusively enriched in synovial tissues but not in blood. Highly suppressive, CD25hi Tregs presented dynamic differentiation potential through high shared clones across different clusters. Computational and in vitro analyses revealed that cortisol induced AREG expression, suppressed glycolysis, and impaired the suppressive function of CD25loAREGpos Tregs. In contrast, CD25hiCXCR6pos Tregs were highly suppressive and showed coordinated abundance with macrophages in synovial tissue which promoted their functional suppressive state through membrane-bound TNFα signaling. These two Treg subsets were similarly found in the synovial tissue in JIA, indicating conserved mechanisms across arthritic diseases. Conclusion Together, our findings define distinct pathways driving divergent functional and dysfunctional Treg states in inflamed tissues and point to interventions that may prevent or reverse the development of the dysfunctional state. Funding Source This work was supported by NIH grants P01AI148102 to MBB, BK was supported by NIH grant T32AR007530-35. SGO was supported by NIH grant (P01AI148102), the Gordon and Llura Gund Foundation and Israeli Council of Higher Education. RS was supported by Mitsubi Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)
The membrane distal domain of CD16a controls NK cell ADCC activity independent of IgG binding 2309237
Abstract Introduction Antibody dependent cellular cytotoxicity (ADCC) is a crucial immune function mediated by natural killer (NK) cells to destroy malignant or virally infected cells. ADCC occurs when IgG opsonized antigens on target cell surfaces bind to the membrane proximal domain 2 (D2) of FcgRIIIa (CD16a). However, the first membrane distal domain 1 (D1) of CD16a has no known function. Methods Here we used cryo-electron microscopy (EM), super-resolution nanoscopy, and functional assays to study the molecular basis of CD16a D1 activity in the context of ADCC. Results We show that the nanobody C28 potently inhibits NK cell ADCC independently of IgG binding while a similar nanobody, C21, does not. C28 does not alter natural cytotoxicity but quenches phosphorylation of the CD16a co-receptor CD3-zeta and alters synapse structure. Cryo-EM analysis of these nanobodies at better than 4Å resolution defines a new CD16a D1 epitope that specifically influences NK cell ADCC. We are currently determining how perturbation to this epitope alters CD16a orchestration during ADCC using MINFLUX nanoscopy with single nanometer localization precision. Conclusion Here we describe a new function of the IgG receptor in self-regulating NK cell ADCC. Future studies aim to identify possible cis-acting proteins ligands underlying this novel function. Using our structure as a blueprint, we are engineering new variants of CD16a D1 that we predict will enhance ADCC activity without affecting natural cytolytic propensity. Such “super-ADCC” NK cells could augment monoclonal antibody therapies for patients with poor response to treatment. Funding Source NIH/NIAID R01 AI167646 Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)