Browse Articles

New‐Era Polymer Thermoelectrics: Material Innovations, Doping Frontiers, Decoupling Strategies, and Unconventional Applications

ABSTRACT The field of polymer thermoelectrics has undergone transformative development in recent years, marked not only by addressing the conventional performance disparity between p‐type and n‐type polymers, but also by innovations in doping methodologies, new strategies for suppressing dopant‐induced disorder, approaches for overcoming the inherent thermoelectric trade‐off, and advancements in achieving better ambient and thermal stability, as well as their variety of new applications. Collectively, these pivotal advances have brought the field into a new era. This review focuses on the recent development of high‐performance polymers that bridge the long‐standing performance gap, introduces doping frontiers for more highly optimized thermoelectric properties, and discusses sophisticated strategies for decoupling electronic and thermal transport. Moreover, this work unlocks the emerging understanding of the degradation mechanisms of doped polymers that enables the design of materials with superior ambient and thermal robustness, and showcases the burgeoning applications of thermoelectric polymers in unconventional domains.

A YafK family protein from Legionella pneumophila exhibits serine-dependent β-lactam hydrolysis activity

Octa-arginine-modified calcium phosphate nanoparticles with amiloride enhance gene delivery and bone augmentation in rat calvaria

Resolvin E1 limits IFNg production by NK cells during SAA 2309893

Abstract Introduction Severe aplastic anemia (SAA) is a bone marrow (BM) failure syndrome resulting from hematopoietic stem cell (HSC) loss driven by cytotoxic T cells. Natural killer (NK) cells were shown to suppress cytotoxic CD8+ T cells via NKG2D, therefore limiting HSC cell death. However, NK cells also exhibit cytotoxicity during SAA and may sensitize HSCs to T cell killing via IFNg. These data suggest that NK cells play a dual role in SAA pathogenesis, with the capacity to both limit and contribute to HSC loss. Resolvin E1 (RvE1) is a pro-resolving lipid mediator previously shown to improve outcomes in a murine model of SAA. NK cells express RvE1’s receptor ChemR23 and RvE1 has been shown to upregulate NKG2D on NK cells in allergic inflammation. To explore mechanisms of RvE1-mediated protection in SAA we aimed to determine if RvE1 influenced NK function in the BM. Methods We utilized a murine model of SAA and published single-cell RNA-sequencing datasets generated from healthy controls and SAA patients. Results In murine SAA, NK cells were elevated in the BM and a greater proportion were cytotoxic. ChemR23 expression was increased on BM NK cells in SAA in mice and human patients. ScRNA-seq generated from SAA mice treated with RvE1 showed increased Nkg2d but reduced Ifng mRNA expression in NK cells compared to vehicle controls, correlating with survival and improved outcomes. Conclusion These data suggest that RvE1 regulates NK function, refining our mechanistic understanding of how RvE1 improves SAA outcomes. Funding Source n/a Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)

STING primes stress granule assembly at the ER 2328483

Abstract Introduction Stress granules (SGs) are stress-induced ribonucleoprotein condensates that are implicated in diverse diseases, including amyotrophic lateral sclerosis (ALS). How SG nucleation is primed prior to stress remains unclear. STING is an ER-resident adaptor known for DNA-sensing innate immune signaling. Analysis of STING interactomes revealed its associations with SG proteins, prompting us to investigate whether STING regulates SG formation. Methods SG assembly in wild-type and STING-deficient cells exposed to oxidative, heat, or ER stress was quantified by immunofluorescence. STING-associated interactions were examined using co-immunoprecipitation, proximity ligation assays, and TurboID-based proximity labeling. Domain mapping was performed using mutants of STING and the SG protein G3BP1. We assessed the role of ER tethering in SG pre-condensate formation, using a construct that anchors the G3BP1-interacting region of STING to the ER without its transmembrane domains. The impact of STING on ALS-associated TDP-43 mutant phenotypes was evaluated. Results Loss of STING reduced SG number and size independently of canonical STING signaling. STING interacted with the SG core proteins G3BP1 and UBAP2L under basal conditions, indicating the presence of ER-associated SG pre-condensates. Expression of an ER-anchored STING C-terminal domain restored SG formation and reduced stress-induced cell death in STING-deficient cells. STING also promoted cytoplasmic accumulation and aggregation of ALS-associated mutant TDP-43 and exacerbated mitochondrial dysfunction. Conclusion STING functions as an ER-resident scaffold that primes SG maturation by pre-organizing core components before overt stress, independently of innate immune signaling. This STING—SG axis links membrane scaffolding to condensate biology and provides a mechanistic connection to neurodegenerative pathology. Targeting STING interactions with SG proteins may offer a therapeutic strategy for diseases driven by aberrant SG assembly. Funding Source n/a Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

A STING-agonist Conjugate Vaccine Potently Synergizes with admixed QS-21 liposomes and CpG to Elicit Robust Th1 Immunity against Chlamydia in mice 2257260

Abstract Introduction Chlamydia trachomatis (CT) is the most common bacterial sexually transmitted infection and can cause pelvic inflammatory disease and infertility in women. CT vaccine development has been hindered by a lack of safe adjuvants that elicit high frequencies of IFNγ memory CD4 T cells (Th1) required for protection. Our lab developed a novel subunit vaccine consisting of the immunodominant Chlamydia protease-like activity factor (CPAF) antigen conjugated to a novel STING agonist CL1151 (CPAF-CL1151), that induces a protective Th1 response in mice. We hypothesized the addition of QS-21 liposomes and/or a TLR9 agonist (CpG1826) with CPAF-CL1151 would synergistically enhance Th1 immunity against Chlamydia. Methods C57BL/6 mice were immunized intramuscularly (i.m.) or via combined i.m. and intranasal (i.n.) route with different adjuvant combinations (-CL1151+QS-21+CpG). CPAF-specific T cell responses were determined by IFNγ ELISpot, ELISA, and ICS. THP-1 derived macrophages were stimulated with CPAF-CL1151, QS-21, or both and cytokine secretion quantified by ELISA. Results ELISpot showed mice vaccinated i.m with -CL1151+QS-21 had a 2-fold increase in splenic IFNγ CD4 T cells compared to -CL1151 (p < 0.001), and an 8-fold increase compared to QS-21 (p < 0.0001). The triple-adjuvant vaccine (-CL1151+QS-21+CpG) delivered i.m + i.n enhanced the frequency of IFNγ CD4 T cells by 2-fold (p < 0.01) and induced a 6-fold increase in IFNγ secretion by splenocytes upon CPAF restimulation (p < 0.01) over -CL1151 + QS-21. In THP-1 macrophages, -CL1151 + QS-21 had >10-fold increases in IFNβ, IL-6, and IL-1β secretion relative to individual adjuvants, indicating activation of interferon and inflammasome pathways. Conclusion These data highlight the Th1 synergy of TLR, saponin, and STING adjuvants and demonstrates the potential of multi-adjuvant vaccines against intracellular bacterial pathogens. Future studies will define the synergistic molecular and cellular pathways underlying this protective CD4 T cell immunity. Funding Source UO1AI182180 Topic Categories Vaccines and Immunotherapy (VAC)

Mechanisms of humoral sensitization to fetal antigens during pregnancy 2259768

Abstract Introduction Sensitization to non-self human leukocyte antigen (HLA) is a barrier to transplant tolerance and graft survival. During pregnancy in humans, exposure to paternal HLA expressed by the semi-allogeneic fetus can induce the production of anti-HLA antibodies and memory B cells despite spontaneous T cell tolerance. Although murine studies suggest that pregnancy elicits B cell responses distinct from those in transplant rejection, the mechanisms driving humoral sensitization to fetal antigens remain poorly defined. Methods To investigate the mechanisms that underlie humoral sensitization in pregnancy, we modeled semi-allogeneic pregnancy by mating C57BL/6 female mice to BALB/c males expressing the model antigen 2W1S-OVA (2W-B/c) to enable the identification of fetus-specific T cells. Fetus-specific T and B cells were identified by flow cytometry using MHC tetramers. Results We found that post-partum mice developed fetus-specific serum IgG1 and IgG2c and fetus-specific IgG-secreting cells in the spleen and bone marrow. The numbers of fetus-specific B cells increased in uterus-draining lymph nodes. To test if fetal MHC-specific memory B cells formed during pregnancy, post-partum mice were challenged with 2W-B/c splenocytes (s.c.), and MHC-specific antibody responses were measured using beads coated with BALB/c MHC class I (Kd) or class II (IEd). Mice mated with allogeneic 2W-B/c males exhibited a more rapid and robust MHC-specific antibody response than mice mated with syngeneic (B6) males, confirming that exposure to fetal antigens during pregnancy elicits the formation of MHC-specific memory. Conclusion These results demonstrate that semi-allogeneic pregnancy sensitizes the maternal immune system to fetal antigens, resulting in the generation of fetus-specific IgG-producing cells and memory B cells despite tolerogenic T cell responses to the fetus. We are investigating the nature of the help provided by CD4+ T cells to fetus-specific B cells and the quality of the memory B cells generated by pregnancy. Funding Source 5R01AI182097 Topic Categories Mucosal and Regional Immunology (MUC)

Siglec-9 is a functional inhibitory receptor for B7-H3 that regulates anti-tumor immunity 2260091

Abstract Introduction Checkpoint blockade revolutionized cancer therapy. However, only a small proportion of cancers respond to checkpoint blockade. B7-H3 is a member of the B7 superfamily, which includes both co-inhibitory and co-stimulatory receptors. B7-H3 is upregulated by many tumors which is generally associated with a worse prognosis. However, the exact mechanism by which B7-H3 suppresses the immune response remains elusive. Methods We transfected human HEK293T cells and assessed the binding to recombinant Siglecs via flow cytometry. We generated monoclonal antibody F7 against human B7-H3 and tested its blocking function using B7-H3 transfected HEK293T cells. We examined the extent of degranulation and NK cell-mediated killing on B7-H3 expressing HEK293T cells in vitro, as well as the CART19 cells mediated killing in relapse model of B7-H3 expressing CD19+ leukemia Nalm6 cells NSG mice. Results We demonstrated that sialylated B7-H3 binds to the inhibitory receptor Siglec-9 on immune cells but not to the related protein Siglec-7. A high affinity monoclonal antibody (mAb) F7 against B7-H3 blocks Siglec-9 binding. Functionally, F7 enhances NK cell killing of B7-H3 expressing target cells in vitro. F7 also dramatically enhances CART19 mediated killing of B7-H3 expressing Nalm6 cells in a relapse model in NSG mice in vivo. Conclusion We have now shown for the first time that B7-H3 binds to the inhibitory receptor Siglec-9 and generated a novel monoclonal antibody F7 that target a unique sequence in B7-H3 as compared to other monoclonal antibodies being examined for clinical efficacy. F7 can promote NK cell killing of B7-H3 expressing target cells in vitro and synergize with CART19 to enhance killing of B7-H3 expressing tumor cells in vivo. As B7-H3 is overexpressed in many human cancers, this work is critical toward understanding how B7-H3 mediates immune evasion through Siglec-9 for enhancing anti-tumor immunity. Funding Source R01CA243545-01A1 Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Targeting lung tissue resident immunity via intramuscular vaccination for protection against Mycobacterium tuberculosis 2300551

Abstract Introduction An improved tuberculosis (Tb) vaccine is needed to confer long lasting protection for all individuals targeting correlates of protection (CoP). We seek to generate a parenterally administered vaccine that induces strong lung-resident immune responses to protect against Mtb. Methods We hypothesize that intramuscular (IM) vaccination with adjuvant double mutant Escherichia coli heat-labile toxin (dmLT) and a bacterial OMV-derived adjuvant (T-vant) will induce a robust response, driving tissue resident immunity in the lung. Mice will be vaccinated IM with antigen alone or antigen with the cocktail in a prime-boost fashion. Two weeks post boost, mice will be intravenously injected with CD45 antibody to differentiate circulating vs non-circulating tissue resident cells. Lymph nodes, spleens, and lungs will be stained with antigen-specific MHCII tetramer to identify CD4+ T cells that are vaccine-specific. Flow cytometry will identify cells that are tissue resident (IV CD45-CD69+P2X7R+), and serum and BAL fluid will be collected for antibody titers. Results In comparison to other routes, IM vaccination generated the highest number of vaccine specific CD4+ and CD8+ T cells. Following IM immunization, we observed significantly higher numbers of tissue resident vaccine-specific CD4+ in the lung of adjuvant vaccinated mice than naïve or antigen alone groups. These cells preferentially displayed a TH1/TH17 phenotype, which are key correlates of protection for Mtb infection. Conclusion These findings suggest that our combination adjuvanted vaccine might produces lung-resident immune responses that could provide increased protection against lung infections. We hope to utilize this in developing novel vaccines for Tb that target CoPs for lasting protection for all individuals. Funding Source NIH Topic Categories Mucosal and Regional Immunology (MUC)

RIG-I promotes protective type I interferon production by glia in response to Neisseria meningitidis and Streptococcus pneumoniae challenge 2257066

Abstract Introduction Bacterial meningitis is a rapidly progressing and often fatal infection of the central nervous system (CNS), driven by glial cell activation and potent neuroinflammatory responses. While many pattern recognition receptors are well characterized in CNS immunity, the role of cytosolic nucleic acid sensors such as retinoic acid-inducible gene I (RIG-I) remains largely undefined. Although classically associated with antiviral defense, emerging evidence indicates that RIG-I can also detect bacterial nucleic acids. Methods To investigate the role of RIG-I during bacterial meningitis, primary murine microglia, astrocytes, and human microglia were infected with Neisseria meningitidis or Streptococcus pneumoniae in the absence or presence of RIG-I-targeting siRNA, recombinant IFN-β, or RIG-I agonists. Whole-cell lysates and supernatants were analyzed for RIG-I and interferon-stimulated gene (ISG) expression by Western blot, cytokine levels (IL-6 and IFN-β) by ELISA and bacterial burden by plating viable CFU. Results We demonstrate that RIG-I is constitutively expressed in human and murine glial cells and is upregulated upon bacterial infection, with protein levels varying by pathogen. RIG-I activation enhanced IFN production and ISG induction, leading to reduced bacterial burden in an IFNAR-dependent manner. Treatment with RIG-I agonists further amplified these protective responses. Additionally, DNA-templated silver nanoclusters (DNA-AgNCs) served as a complementary antimicrobial approach, exhibiting potent bactericidal activity against N. meningitidis and S. pneumoniae in infected glia. Conclusion Together, these findings identify RIG-I as a key cytosolic sensor driving protective IFN-dependent antimicrobial responses in glial cells, while AgNCs offer a direct antimicrobial strategy. Collectively, these approaches highlight the potential for targeting both innate immune pathways and direct antimicrobial mechanisms to enhance pathogen control during bacterial meningitis. Funding Source R03 NS120047-01A1 Topic Categories Neuroimmunology (NEUR)

BACH2 dosage establishes the hierarchy of stemness and finetunes antitumor immunity in CAR T cells 2256626

Abstract Introduction Self-renewing stem-like T cells promote the efficacy of cancer immunotherapy and are a heterogeneous population with sub-lineages demonstrating different degrees of stemness. At the apex of this hierarchy are long-term (LT) stem-like T cells with the highest capacity of persistence, repopulation and response to immune checkpoint inhibitors (ICI). However, the pathway that establishes the hierarchy of stemness in chimeric antigen receptor (CAR) T cells and its role in anti-tumor efficacy of CAR T cells are unclear. Methods We analyzed CAR T cell infusion products from both humans and mice, examining epigenetic, transcriptional, and functional correlates of stem-like subsets. We deleted BACH2 to assess its role in LT stem-like differentiation and antitumor potency. In a GD2 CAR T model prone to exhaustion, we used small-molecule modulation to fine-tune BACH2 protein levels and temporally activate BACH2 during CAR T manufacture, followed by in vivo efficacy testing in solid tumor models. Results We demonstrate that BACH2 regulates LT stem-like CAR T cell differentiation and antitumor immunity in a dose-dependent manner. Pre-infusion LT stem-like CAR T cells displayed epigenetic activation of BACH2 and correlated with more robust antitumor responses in both mice and humans. After tumor clearance in vivo, LT stem-like cells emerged that transcriptionally and epigenetically upregulated BACH2 and downregulated TOX. Loss of BACH2 impaired antitumor efficacy and disrupted the LT stem-like transcriptional program. In GD2 CAR T cells, quantitative control of BACH2 via a small molecule adjusted the balance between stemness and exhaustion. Temporal induction of BACH2 during CAR T manufacturing enhanced antitumor immunity in solid tumor models compared to unmodified cells. Conclusion Together, we show that BACH2 dosage establishes the hierarchy of stem-like CAR T cells and can be temporally and tunably controlled in CAR T cells to optimize differentiation and antitumor immunity. Funding Source AI154450 from National Institutes of Health, RR210035 from Cancer Prevention and Research Institute of Texas, HT94252310801 from Department of Defense to C. Yao; AI158294, AG083398, AG056524 from National Institutes of Health, Clinic & Laboratory Integrat Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Dual‐Functional Fluorinated Additive in Gel Polymer Electrolyte for High‐Energy‐Density Anode‐Free Lithium Batteries With Enhanced Thermal Stability

ABSTRACT Anode‐free lithium metal batteries offer high energy density and low cost, but their practical deployment is limited by unstable lithium–electrolyte interfaces and safety risks from dendrite growth and flammable electrolytes. Here, we report a dual‐functional fluorinated additive that enables in situ gelation, forming a thin, inorganic‐rich solid electrolyte interphase while simultaneously enhancing flame retardancy through radical capture. At 0.5 C discharge, the additive‐enabled anode‐free pouch cell delivers high specific capacities of 202.1 mAh/g (3.0–4.5 V) and 192.0 mAh/g (3.4–4.3 V), while retaining 80.3% and 80.1% of their initial values after 85 and 100 cycles, respectively. Moreover, Cu/NCA pouch cells with capacities of 1.4 Ah (3.0–4.5 V) and 200 mAh (3.6–4.3 V), incorporating the additive, retain 80.2% of initial capacity after 85 cycles and 92.9% after 100 cycles, respectively. Such performance is on par with or better than the best results in the literature. Impressively, such a 1.4 Ah cell shows no thermal runaway in a harsh drilling test at the fully charged state, even after 100 cycles. These results demonstrate that the dual‐functional fluorine additive leads to both excellent electrochemical performance and enhanced safety, paving the way for the safe and practical application of anode‐free lithium metal cells.

High Piezoelectricity and Temperature Stability via Stabilized Polar Distortion

ABSTRACT Simultaneously combining high piezoelectric performance with excellent thermal stability is essential for piezoelectrics operating under high‐temperature conditions, yet these two attributes are often in competition. Here, we propose a design strategy that stabilizes the intrinsic lattice contribution by constructing a mixed‐symmetry ferroelectrically distorted state and demonstrate its effectiveness in Pb(Zr 0.53 Ti 0.47 )O 3 ‐ x Nb (N x ) ceramics. The optimized N3 composition exhibits a high piezoelectric coefficient d 33 of 550 pC/N and a high Curie temperature T C of 367°C. Over the wide temperature range of 25–300°C, the variations in piezoelectric coefficient ( d 33 ) and electromechanical coupling factor ( k p ) are limited to only 6% and 9%, respectively. In situ temperature‐dependent structural analyses reveal that the enhanced piezoelectricity and thermal robustness originate from a ferroelectric distortion that is strongly developed at room temperature due to niobium doping and remains stable up to 300°C, as further corroborated by first‐principles calculations and scanning probe microscopy measurements. This mixed‐symmetry‐stabilization strategy provides a generalizable route to overcoming the conventional trade‐off between performance and stability and offers design guidelines for next‐generation high‐performance piezoceramics tailored for high‐temperature applications.

Tough, Ductile, and Strong Hard‐Soft Cementitious Composite Enabled by Multi‐Material Additive Manufacturing

ABSTRACT Monolithic cementitious materials lack fracture resistance and are brittle. Advancements in additive manufacturing techniques with cement‐based materials and architected designs have remained limited to the use of a single (cement‐based) constituent. This work charts a new pathway for manufacturing and designing tough, ductile, and strong architected cement‐based composites by proposing a novel multi‐material additive manufacturing (MMAM) technique for the first time, integrated with a coupled experimental‐numerical design approach. The new class of architected cementitious composites (ACC) of mortars and elastomeric constituents (silicone and polyurethane) are exemplified, through a layered hard‐soft architected design, inspired by the microstructure of a sea sponge (glass sponge Euplectella aspergillum ). The MMAM technique alternates extrusion of hard‐soft composites, enabling systematic control over the geometry and constituents of resulting architected structures. The results demonstrate layered mortar‐silicone composites achieved up to 3.9‐ and 8.8‐fold enhancements in fracture toughness and up to 11.7‐ and 12.4‐fold enhancements in ductility relative to monolithic 3D‐printed (3DP) and cast mortars, respectively. A coupled large‐deformation phase‐field–cohesive‐zone (PF‐CZM) framework was used to systematically probe soft‐layer thickness and bulk soft‐material properties. Simulations revealed that combining higher‐stiffness soft layers with reduced thickness can yield up to a 24‐fold increase in work‐of‐fracture while recovering the load‐bearing capacity of monolithic mortar. Guided by numerical predictions, experiments on thin, stiffer polyurethane interlayers validated the numerical predictions and provided additional experimental evidence that the proposed (mortar‐polyurethane) composites achieve 82‐ and 187‐fold higher fracture toughness, and 22.6‐fold higher ductility, relative to 3DP and cast monolithic references while recovering the flexural strength to levels statistically comparable to monolithic mortar. These large gains arise from three synergistic mechanisms: crack arrest/deflection, crack bridging, and discontinuous layerwise crack re‐nucleation, activated by the layered hard‐soft architecture and supported by DIC/AE fracture analyses). The proposed MMAM‐enabled fabrication‐design‐mechanics approach in ACC can unleash entirely new pathways for engineering next‐generation damage‐resilient and multi‐functional concrete structures.

Structure-based virtual screening identifies potential endogenous ligands of the human bitter taste receptor TAS2R46

Prevalence of cervical human papillomavirus infections and cytological abnormalities in asymptomatic women living with HIV in Addis Ababa, Ethiopia

Abstract Cervical cancer from persistent HPV infection is the leading cause of cancer death among Sub-Saharan African women. We here addressed the existing knowledge gap concerning the prevalence of HPV infection and cervical lesions in asymptomatic women living with HIV (WLWH) in Addis Ababa, Ethiopia. Adult WLWH ( N  = 334) were recruited at the HIV clinic of the Tikur Anbessa Specialized Hospital, Addis Ababa. Real-time PCR targeting 12 high-risk (HR) and 2 low-risk HPV types on cervical samples and Pap smear testing were performed. Logistic regression was used in IBM SPSS Statistics to identify factors associated with HPV infection and abnormal cytology. HR-HPV infections occurred in 20.6% of participants and the most prevalent HR-HPV genotypes were HPV35 (16.8%) and HPV16 (14.7%). Abnormal cytology was more common among HR-HPV–positive women than HR-HPV–negative women (36.1% vs. 12.6%, p  < 0.0001). LSIL and HSIL were detected in 9.9% and 6.6% of HR-HPV–positive women, respectively. Condom use was associated with reduced odds of HPV infection and abnormal cytology. HR-HPV infections dominated by non-HPV16/18 types and abnormal cytology were common among asymptomatic WLWH in Ethiopia underscoring the need for strengthened cervical cancer screening, HPV vaccinations and safe-sex education for this high-risk population.

Myeloid-specific NLRP12 is a checkpoint of colonic inflammation by promoting taurine-driven microbiota 2309979

Abstract Introduction Inflammatory bowel diseases (IBD) exhibit a global disease burden, with pathogenesis driven by the dynamic interplay of genetic susceptibility, gut microbial dysbiosis, and immune dysregulation. We studied the tissue-specific regulatory role of Nlrp12 in gut microbial dysbiosis and colonic inflammation and revealed a complex regulatory link of myeloid-specific NLRP12 and the promotion of taurine to induce colitis-mitigating bacteria while restraining bacterial species that exacerbate gut inflammation. Methods We generated Nlrp12flox/flox (Nlrp12fl/fl) mice and created both myeloids-specific cre and epithelial-specific cre mouse. We used both conventional mouse and germ-free mouse model. For acute colitis induction, we used DSS for 5 days and then regular drinking water. DSS-AOM was used for colorectal cancer model. ELISA and western blot were used to study cytokine production and signaling pathways. Fecal microbiome profiling was done by 16s rRNA sequencing. We also have human fecal sample and qPCR was used to quantify specific strains. Fecal microbiome transfer was used to study the causality between genetics, microbial species and colitis. Results Myeloid-specific , but not epithelial-specifc expression of NLRP12 is protective against colitis. Deficiency of Nlrp12 in myeloid cells has minimal effects on colorectal cancer. Myeloid-specific NLRP12 does not result in inflammasome activation in the colitis model. Deficiency of Nlrp12 in myeloid cells promotes microbial dysbiosis. Fecal microbiota transplantation from WT mice attenuates colonic inflammation in Nlrp12ΔMye mice. Allobaculum exacerbate DSS-induced colitis in GF mice. Human IBD patient samples demonstrate similar microbial signatures. Fecal metabolomic analysis reveals alterations in taurine and bile acid metabolism in Nlrp12ΔMye mice. Conclusion This work reveals the profound effect of a host innate immune checkpoint that functions in a cell-specific fashion to protect against metabolomics changes and dysbiosis. that leads to colitis. Funding Source NIH Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Target-specific monoclonal antibodies detect the GPCRs PAR1 or DRD2 expressed on live cell surfaces by Flow Cytometry 2260881

Abstract Introduction G protein-coupled receptors (GPCRs) are the largest family of multi-transmembrane proteins, and major targets for drug discovery. Antibody staining/detection of GPCR targets on live cells is important for studying receptor expression, localization, and dynamics in their native, functional state. Due to their complex structure and dynamic conformations, it is very challenging to develop antibodies against GPCRs. Protease-activated receptor-1 (PAR1) and Dopamine receptor D2 (DRD2) are Class A GPCRs. PAR1 is a promising target for cancer therapeutics, while DRD2 is a pivotal target for therapeutic intervention in the treatment of neuropsychiatric disorders. Methods Based on multiparameter analyses of PAR1 and DRD2 sequences and structures, we designed unique immunogens and immunized mice. After detection of PAR1 or DRD2 antibodies in immunized mouse sera, immune mouse B cells were isolated to generate PAR1- or DRD2-specific monoclonal antibodies (mAbs) using hybridoma technology. The PAR1- or DRD2-mAbs were characterized for binding specificity, affinity and live cell staining using multiple assays. Results Using unique fit-for-target immunization platforms, PAR1 or DRD2 specific antibodies were elicited in mice that could bind to the target antigen by ELISA and target protein expressed on live cell surface by flow cytometry. Subsequently, PAR1 or DRD2 specific mAbs were developed from immunized mouse B cells. PAR1- or DRD2-specific mAbs had high binding specificity and affinity to the relevant target antigen as measured by ELISA and surface plasmon resonance (SPR). Flow cytometry analyses demonstrated that the mAbs could potently bind to PAR1- or DRD2 expressed on live cell surfaces. Conclusion As antibody staining of GPCRs on cell surface is a foundational and indispensable technique, these PAR1 and DRD2 specific, potent and live cell binding mAbs can be very useful tools for PAR1 and DRD2 research and drug discovery. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)

Inhibition of Glutamine metabolism is essential in decreasing the hallmarks of Atopic Dermatitis pathogenesis 2257318

Abstract Introduction Atopic Dermatitis (AD) remains one of the most chronic inflammatory skin diseases and is strongly associated with other atopic diseases like asthma. Though Th2 response is known to be the main humoral response in AD, follicular helper T (Tfh) cells have been recently identified as key mediators in AD pathogenesis. AD presents with increased serum IgE levels and associated cytokines, IL4 and IL13. Our lab has shown that upon stimulating Dendritic cells with Altenaria alternata (a fungal allergen) in an allergic asthma model, there was an increase in the glutamine metabolic pathway. Upon inhibiting the glutamine metabolic pathway, we observed a reduction in downstream Tfh2 polarization and reduced IgE production. We therefore hypothesized that Telegnastat (CB-839), a glutaminase I inhibitor would decrease Tfh2 responses in AD. Methods Auricles of C57BL/6 mice were treated with MC903 and house dust mite for a period of 15 days. Ear thickness was measured throughout the model. Mice received topical treatment with CB-839 after day six. Mice were euthanized on day 15; organs were harvested and processed for flow cytometry, sectioning, RNA isolation and bulk RNA sequencing. Results Topical treatment with CB-839 inhibited AD in treated mice with a reduction in IgE+ and IgG1+ Plasma cells as well as Tfh2 cells, while an increase in T follicular regulatory cells was observed. AD associated genes such as GATA3 and CARD11 were observed to be decreased in CB-839 treated mice. Bulk RNA sequencing analysis also showed that with treatment, there was an increase in genes involved in wound healing, immune regulation and tissue development. Conclusion Overall, topical treatment of AD with the GLS-1 inhibitor, CB-839 in mice is associated with a decrease in the hallmarks of AD. Funding Source NHLBI-NIH (R01HL162991) to RKM Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)

Rapid protection from lethal Orthopoxvirus infection following vaccination with mRNA-LNP encoding the Type I Interferon Binding Protein 2259797

Abstract Introduction Orthopoxviruses (OPVs), such as monkeypox, pose ongoing public health threats, especially since the global herd immunity has declined following the cessation of routine smallpox vaccination. Current vaccine strategies have limitations due to safety and short-lived immunity. Most anti-viral vaccines aim to induce antibodies (Abs) that bind to structural viral proteins, preferentially those that can neutralize the viral particle. The Type I IFN-binding protein (IFN-I bp) is a highly conserved non-structural protein present in all OPVs, an immune evasion protein, and is essential for their pathogenicity. It has been previously shown that mice vaccinated with recombinant IFN-I bp or passively immunized with anti-IFN-I bp monoclonal antibodies (Abs), are protected from lethal challenge with the mouse OPV ectromelia virus (ECTV), a classic model for human smallpox and monkeypox infections. Methods We made mRNA-lipid nanoparticle (mRNA-LNP) vaccine encoding IFN-I bp. We evaluated Abs responses by ELISA, and protective efficacy by challenging the immunized mice with ECTV in the footpad or with the OPV vaccinia virus intranasally. Results We found that a single immunization of 5 ug of mRNA-LNPs encoding ECTV’s IFN-I bp induced much higher titers of IgG antibodies (as opposed to IgM) as early as six days post-immunization than control mRNA-LNP vaccines. Notably, vaccinated BALB/c and TLR9-deficient mice challenged with ECTV or VACV eight days post-immunization were fully protected from viral lethality. Conclusion These data indicate that the IFN-I bp is a promising antigen for OPV vaccines, particularly when rapid protection is crucial, such as during an epidemic. Ongoing studies aim to determine the mechanisms underlying rapid IgG induction. Funding Source grant Topic Categories Vaccines and Immunotherapy (VAC)