Browse Articles
Discover research articles across all indexed journals
De novo generation of innate memory-like T cells occur early during human immune development 2254423
Abstract Introduction Infants rely heavily on innate immunity while adaptive memory develops. A subset of T cells, termed “virtual memory” or innate-like memory T cells, bridges innate and adaptive immunity, but their emergence in early human life is unclear. Infants receiving gene therapy for X-linked severe combined immunodeficiency (SCID-X1), whose immune systems reconstitute de novo, offer a unique model to study this process. We investigated whether innate-like memory CD8+ T cells arise during immune reconstitution and defined their epigenetic, transcriptional, and functional features. Methods Longitudinal blood samples from SCID-X1 infants post—gene therapy were analyzed. Flow cytometry identified CD8+ subsets expressing NKG2A. TCR sequencing assessed clonal diversity, while DNA methylation and RNA profiling defined molecular signatures. Functional assays measured cytokine-induced, antigen-independent IFN-γ production after IL-12 and IL-18 stimulation. Results Early after gene therapy—induced immune reconstitution, infants’ peripheral blood showed a relative enrichment of innate-like memory CD8+ T cells marked by NKG2A expression. These NKG2A+ CD8 T cells displayed innate-associated transcriptional signatures and a TCR repertoire distinct from conventional naïve/memory subsets. DNA methylation profiling confirmed a subset-specific epigenetic signature, including open (poised) regions at effector genes, consistent with readiness for rapid response. Functionally, when stimulated ex vivo with IL-12 plus IL-18, NKG2A+ T cells produced IFN-γ in an antigen-independent fashion, indicative of innate-like responsiveness. Together, these features distinguish them from conventional antigen-driven memory CD8 T cells. Conclusion Innate-like memory CD8+ T cells emerge early during immune reconstitution, primed for rapid, antigen-independent responses. Funding Source NIH(R01AI114442 ; R01CA237311 ; P01-HL053749; U54AI082973;P01AI138962), The ASSISI Foundation ofMemphis, AlexLemonade Stand, SU2C, Key for a Cure Foundation, NRSA(F32-CA250155), CIRM (CLIN2-10830;CLIN2-09504), and theAmerican Lebanese Syrian Associated Ch Topic Categories Hematopoiesis and Immune System Development (HEM)
Nitrogen‐Incorporated Silicon Dioxide Interlayer Enables Pinhole‐Reduced and Robust TOPCon With a High Implied Open‐Circuit Voltage over 760 mV
ABSTRACT Tunnel oxide passivating contact (TOPCon) technology has emerged as an industrial cornerstone for high‐efficiency crystalline silicon (c‐Si) solar cells. However, its passivation performance remains constrained by the thermal fragility of the ultrathin silicon dioxide (SiO x ) interface and the resulting formation of pinholes. Here, we report a nitrogen (N)‐incorporated TOPCon structure in which nitrogen (N) atoms are in situ doped into amorphous silicon via plasma‐enhanced chemical vapor deposition (PECVD) and are driven to the SiO x during annealing to form a robust SiO x N y interlayer. First‐principles calculations reveal that SiO x N y exhibits significantly enhanced bonding strength and superior thermal stability. Finite element simulations further show that SiO x N y possesses a thermal expansion coefficient better matched to both c‐Si and poly‐Si, suppressing stress concentration, preventing interface fracture, and reducing pinhole density. This atomic‐scale modification enables record‐breaking passivation performance, achieving an implied open‐circuit voltage of 760 mV and a low single‐sided recombination current density of 0.35 fA/cm 2 . Device simulations predict that the efficiencies of various TOPCon‐based cells can be significantly enhanced with minimal additional cost. Experimental data demonstrate that front‐junction TOPCon cells can achieve an efficiency improvement of 0.2%. This simple and industry‐compatible interface‐engineering strategy provides a highly scalable pathway for boosting TOPCon and back‐contact TOPCon solar cells’ performances.
Imaging Flow Cytometry Enhances Characterization of Immune Cell Phenotype and Function 2255710
Abstract Introduction The addition of cellular imaging to high-speed, single-cell flow cytometry has revolutionized simultaneous immunophenotyping with morphological and spatial insights. Methods We have pioneered the utilization of imaging-enabled platforms such as the CytPix and FACSDiscover to characterize cell biology. The CytPix combines standard flow cytometry with a 20X equivalent brightfield camera to provide detailed morphological information, while the FACSDiscover enables cellular marker localization using three fluorescence imaging detectors in addition to a full spectral cytometer. Results Both platforms process images at ultra-high throughput, with FACSDiscover reaching up to 10,000 cells per second. Imaging-derived features and AI-based analysis provide morphological and spatial information, uncovering insights not possible with traditional flow cytometry. This includes enhanced identification of immune cell subsets with unique functionalities, improved resolution of senescence, and more detailed signaling and metabolic characterization of mitochondrial activity and distribution. Moreover, imaging cytometry allows to characterize immunological synapses, which was previously impossible with flow cytometry. Conclusion Current developments include leveraging AI to identify cellular states and activation label-free, simplifying cellular characterization and profiling, and bridging phenotype with mechanism–thereby driving innovation in immune cell research across multiple therapeutic areas. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)
DUSP22 dysregulation and UBR2 upregulation induce T-cell receptor signaling and systemic lupus erythematosus 2256861
Abstract Introduction Downregulation of DUSP22 in peripheral blood T cells is correlated with human systemic lupus erythematosus (SLE) nephritis. DUSP22 dephosphorylates and inactivates Lck in TCR signaling, leading to suppression of autoimmune disease. We found that DUSP22 dephosphorylated E3 ubiquitin ligase UBR2, leading to its K48-linked ubiquitination and degradation. Remarkably, UBR2 induced Lck K63-linked ubiquitination, leading to Lck Tyr394 autophosphorylation and activation upon TCR signaling. The induction of proinflammatory cytokines and autoimmunity in DUSP22 KO mice was blocked by knocking out UBR2 in vivo. Furthermore, UBR2 protein levels, the UBR2-Lck interaction, and Lck K63-linked ubiquitination were induced in T cells of human SLE patients. Methods We performed NGS deep sequencing to identify DUSP22 gene variants in SLE patients from two independent cohorts, and characterized the functional consequences of the identified DUSP22 variants. Results Our NGS data revealed that 46 DUSP22 gene variants occurred in 20.79% or 18.75% SLE patients from two cohorts. Among the 46 DUSP22 variants, 6 DUSP22 somatic variants altered DUSP22 codons, and another variant was located in the DUSP22 promoter region. Moreover, 16 and 12 variants in the 3’-UTR of the DUSP22 gene were identified from Cohort #1 and Cohort #2, respectively. The functional effects of these identified DUSP22 gene variants were investigated by site-direct mutagenesis, promoter reporter assays, immunoblotting analyses, and 3’-UTR reporter assays. Interestingly, the DUSP22 promoter activity, DUSP22 protein level, or DUSP22 3’-UTR activity was reduced by individual DUSP22 gene variants. Using mass spectrometry-based proteomics, we identified an E3 ligase that induced K48-linked ubiquitination and subsequent proteasomal degradation of the DUSP22 protein. Furthermore, DUSP22 variant-knockin mice indeed spontaneously developed nephritis. Conclusion Collectively, DUSP22 deficiency and UBR2 induction play critical roles in the pathogenesis of SLE. Funding Source n/a Topic Categories Basic Autoimmunity (BA)
Streamlined and cost-effective 96-well plate NGS assay for single-cell immune receptor chain pairing and T/B cell immunophenotyping 2260349
Abstract Introduction To facilitate the discovery of epitopes or immune therapy targets for researchers without ready access to expensive instrumentation or reagents, we present a cost-effective, simplified workflow for single-cell immune receptor (TCR/BCR) profiling. Methods We use a standard 96-well plate; T- or B-cells are flow-sorted into wells, primers for immune receptor and key marker genes are added, followed by multiplex RT-PCR and next-generation sequencing (NGS). Novel validator barcodes (VBCs) ensure robust clonotype quantification with minimal cross-well contamination. There are two workflow options: one can either 1) flow-sort 96 individual cells onto a plate, or 2) serially dilute ∼10,000 cells (∼100 cells/well in a 96-well plate), which allows for thorough characterization of each receptor in larger cell populations. Results Data analysis with MiXCR software yields full-length receptor chain-pair sequences, clonotype abundance, and gene expression profiles. Single-cell results show 86% of the sorted cells had an associated clonotype, 65% had a TCR-αβ chain pair. Conclusion This sensitive assay is ideal for working with rare or enriched cell populations of a few thousand cells. The single-day workflow, which includes pooling of a 96-well plate into one reaction and profiling multiple plates in one batch, drops assay costs to ∼10 cents/cell. The approach will facilitate applications such as engineering T-cell-based therapies, antibody synthesis, and epitope discovery for cancer or autoimmune diseases. Funding Source N/A Topic Categories Technological Innovations in Immunology (TECH)
Combokines: Platform for Cytokine Delivery in aAPC Engineering 2252933
Abstract Introduction CAR-T therapy is promising, but poor T cell expansion causes up to 13% of patients to drop out. Traditional bead-based activation fails to present physiological signals, whereas artificial antigen presenting cells (aAPCs) provide a superior alternative to present membrane-bound proteins for antigen recognition, costimulation, and cytokine stimulation. aAPC engineering is constrained by lentiviral packaging limits and reduced viability after repeated transduction, which restricts the number of introduced signals. To overcome this, a membrane-bound cytokine complex, termed “Combokine”, was developed to present two cytokines simultaneously. Methods Combinations of membrane-bound IL—7/IL-21, and IL-18/IL-21 constructs were tested. K562 cell lines were transduced with lentivirus, single-cell sorted, and validated with flow cytometry. IL-7 and IL-21 activity was confirmed by STAT3/5 phosphorylation using intracellular staining followed by flow cytometry. IL-18 activity was confirmed via NF-kB signaling detected with HEK-Blue IL-18 Reporter cells. Results IL-7/IL-21 Combokines induced STAT3/5 phosphorylation. IL-18/IL-21 Combokines induced STAT3 phosphorylation and activated NF-kB signaling. Combokines with IL-21 in the second position produced greater STAT3 phosphorylation. Similarly, IL-18/IL-21 Combokines with IL-18 in the second position produced stronger NF-kB signaling. Combokine-expressing cell lines induced STAT3/5 phosphorylation and NF-kB signaling comparable to cell lines dual-transduced with IL-7/IL-21 and IL-18/IL-21. Conclusion These findings establish Combokines as a platform to enhance T cell proliferation by integrating multiple cytokine stimulation pathways into a single construct. Future work will evaluate consequences on CAR-T cell proliferation and function. Funding Source The Roy and Diana Vagelos Program in the Molecular Life Sciences Topic Categories Cytokines and Chemokines and their Receptors (CCR)
Hematopoietic stem and progenitor cells overcome treatment resistance to immune checkpoint inhibitors in preclinical gliomas 2240834
Abstract Introduction Glioblastoma (GBM) is one of the most lethal primary brain tumors, with dismal survival. Immune checkpoint inhibitors (ICIs) have revolutionized cancer therapy, with efficacy shown in several solid tumors, yet its impact in GBM has been dismal. To overcome this treatment resistance, we developed a combinatorial strategy incorporating hematopoietic stem and progenitor cells (HSPCs) , whole brain radiation therapy (WBRT), and ICI with either αCTLA-4 or αVISTA in a syngeneic murine glioma model that is completely unresponsive to ICIs. However, our combination therapy significantly prolonged survival in glioma-bearing mice compared to controls. These findings support the potential of multi-pronged immunotherapeutic strategies to improve outcomes in glioma and highlight VISTA and CTLA-4 as promising targets. Methods C57BL/6 mice received orthotopic KR158B gliomas followed by WBRT at 9 Gy single dose. Bone marrow-derived lineage- HSPCs were isolated and co-transferred with ICI, either 100ug loading dose of αCTLA-4 or 30ug αVISTA. Results Our data demonstrated a significant survival benefit using the combinatorial therapy of HSPC + WBRT + αCTLA-4, with over 50% of the glioma-bearing mice surviving beyond 150 days. Similarly, HSPC + WBRT + αVISTA also led to a significant survival advantage compared to control groups. Conclusion The use if HSPCs in combination with ICI remarkably resets the immune system to allow the efficacy of ICI in otherwise treatment resistance gliomas. Funding Source n/a Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)
CD137 and IFN-γ signaling in mature cDC2Bs play a central role in the regulatory feedback mechanism for alloimmune responses 2328469
Abstract Introduction Conventional dendritic cells (cDCs) are a prototype of professional antigen-presenting cells that dictate T-cell activation and differentiation: however, the function of post-primed cDCs is yet to be clarified. Here, we report the identification of a mature subset of type 2 cDCs (cDC2s) capable of suppressing alloimmune responses. Methods Donor BM12(H2-Ab1bm12) or Balb/c splenocytes/lymphocytes (8x10^7) were transfered to C57BL/6 or CBF1 recipients to induce chronic GVHD or acute GVHD, respectively. Various immunological tools were used to analyze immune responses. Results In autoantibody-mediated chronic graft-versus-host disease (GVHD) models, Tnfrsf9 deficiency in recipients resulted in a dysregulation of FCGR1+FCGR2B+CLEC12A+ splenic cDC2Bs, leading to uncontrolled type 1 alloimmune responses. By contrast, engagement of CD137 (encoded by Tnfrsf9) reenforced the immunoregulatory capacity of these inflammatory cDC2Bs in such a way that they suppressed type 2 as well as type 1 alloimmune responses. Mechanically, AHR induced by CD137 signaling suppressed IFN-γ-stimulated genes (e.g., Il12a) by repressing STAT1 transcriptional activation, while upregulating immunoregulatory genes (e.g., Il10) in cooperation with STAT1. IFN-γ promoted production of IL-12 and deletion of donor Treg cells in Tnfrsf9-/- recipients. Conclusion Taken together, this study identifies CD137 signaling in mature cDC2Bs as a master switch for negative feedback regulation mediated by IFN-γ to restrict type 1 alloimmunity-induced systemic inflammation. Funding Source National Research Foundation of Korea (NRF), Republic of Korea Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)
An integrated cell atlas of the innate lymphoid cells in health and disease 2254590
Abstract Introduction Innate lymphoid cells (ILCs) represent a highly diverse family of lymphocytes that play pivotal roles in tissue homeostasis, inflammation, and cancer immunity. Despite growing evidence of their plasticity and functional heterogeneity, a comprehensive understanding of human ILC diversity across pathological contexts remains limited. Methods We performed large-scale single-cell RNA sequencing (scRNA-seq) analysis of 3,322 human samples spanning 60 physiological and pathological conditions. Results By integrating and harmonizing ILC transcriptomes across tissues and diseases, we systematically characterized the cellular states, transcriptional regulators, and disease-specific adaptations of ILC subsets. Our atlas reveals both conserved and disease-specific transcriptional programs of ILCs. Pathological conditions induce profound compositional and phenotypic shifts in ILC subsets. Comparative analyses between TCF7+ ILC1 and ZNF683+ NK-derived IFNG+ ILC1 populations highlight distinct transcription factor dependencies and microenvironmental stimuli shaping their effector programs. Notably, GZMB+ ILC2 cells, enriched in multiple cancer types, were associated with improved patient prognosis, suggesting a potential cytotoxic role. In contrast, NR4A1+ ILC3 cells within NCR⁻ ILC3 clusters were preferentially enriched in tumors and inflammatory diseases, correlating with poor prognosis and resistance to immunotherapy. Conclusion This comprehensive single-cell atlas delineates the dynamic landscape of human ILCs across diseases. Funding Source n/a Topic Categories Computational and Systems Immunology (COMP)
Aromaticity‐Localized Square Tetraboron‐Extended Multiple‐Resonance Emitters With B‒N Covalent Bonds for Highly Efficient Narrowband Electroluminescence
ABSTRACT Polycyclic aromatic hydrocarbons with multiple‐resonance (MR) effect are emerging as leading candidates for wide‐color‐gamut displays. However, it remains a significant challenge to red‐shift their emissions to long wavelengths without sacrificing the spectral full width at half maximum (FWHM). Herein, a synergistic strategy of aromaticity localization and tetraboron extension is newly proposed, in which a B‒N covalent‐bond‐fused MR core is fused with 5,10‐dihydro‐5,10‐diphenylphenazine (DPPA) to afford two yellow‐emitting narrowband tetraboron emitters. By integrating multiple local B–π–B motifs into a rigid square‐like tetraboron MR framework, DPA‐4B and DMAC‐4B achieve a pronounced red shift from violet‐blue DPA‐1B to yellow emission. The antiaromatic N 2 C 4 ring and surrounding nonaromatic 1,2‐BN/1,4‐BN heterocycles promote aromaticity localization and suppress excited‐state relaxation, affording narrow emissions at 544–559 nm with FWHMs of 20–21 nm. The DPA‐4B and DMAC‐4B‐based sensitized OLEDs further achieve narrowband yellow electroluminescence with maximum EQE values of 36.7% and 30.1%, respectively. These results demonstrate that the combination of aromaticity localization and tetraboron extension is a promising molecular design rule for simultaneously realizing emission red‐shifting, spectral narrowing, and device efficiency in B‒N covalent‐bond‐containing MR emitters.
Dissecting transcription factor networks for tissue-resident CD8 T cell differentiation and maintenance 2255891
Abstract Introduction Tissue-resident memory (Trm/TDRM) T cells populate non-lymphoid tissues and provide a first line of defense against pathogen re-exposures. KLF2, Runx3, Hobit, and Blimp1 are transcription factors (TF) that are reported to contribute to Trm/TDRM differentiation or maintenance. Yet it is unclear what the relative importance of each TF is, and how that may vary among tissue microenvironments. Methods We used CRISPR-Cas9-mediated deletion of TFs to address CD8+ Trm/TDRM development in 15 distinct tissues following lymphocytic choreomengitis virus infection (Armstrong strain). Results We found modest contributions by Hobit with few significant changes in knockout Trm/TDRM cells. Runx3 deletion had minimal effects on Trm/TDRM establishment, however integrin expression was significantly altered, including reduced CD103 expression. Blimp1 deletion increased the proportion of recirculating memory T cells that expressed CD62L but had minimal impact on resident cells. Most notably, the loss of KLF2 promoted greater accumulation of Trm/TDRM-phenotype CD8 T cells within the liver and secondary lymphoid organs (SLOs) by seven days after infection and persisted long after antigen clearance. Trm/TDRM-phenotype memory cells expressing CD69, CD49a, and P2rX7 were particularly elevated within SLOs when KLF2 was lost. Conclusion These data indicate that the loss of KLF2 has a major influence on promoting Trm/TDRM establishment, Runx3 shapes Trm/TDRM phenotype, and Hobit and Blimp1 have fewer observable roles after LCMV infection of the parameters analyzed. A better understanding of the TF-mediated regulation of Trm/TDRM will help inform manipulation or programming of T cells for adoptive cell therapies, therapeutic depletion, and other applications. Funding Source n/a Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)
Spatial immunoprofiling of human colon in health and disease: insights into inflammation 2256907
Abstract Introduction Inflammatory bowel diseases (IBD) are chronic, relapsing inflammatory conditions of the gastrointestinal tract. Despite advances in understanding their pathogenesis, challenges remain in diagnosis, treatment, and long-term management [1]. Their clinical heterogeneity, variable therapeutic response, and lack of reliable biomarkers, underscore the need for improved spatial and molecular characterization of immune mechanisms to uncover pathophysiological processes and identify potential therapeutic targets [2]. Here we leveraged automated multiplex sequential immunofluorescence (seqIF™) [3] to spatially profile protein expression and cellular interactions across a spectrum of colon pathologies. Methods A tissue microarray comprising Crohn’s disease, ulcerative colitis, chronic colonic inflammation, neoplastic lesions, and normal colon tissues was stained and imaged on the COMET™ platform. Multiplexed images were analyzed at single-cell spatial resolution using HORIZON™ software. Results The high-plex proteomics panel enabled the simultaneous detection of over 60 biomarkers on the same tissue slide, allowing an in-depth characterization of inflammatory responses and mucosal remodeling associated with inflammatory colon pathologies. This spatial profiling of the tissue microenvironment revealed the cellular heterogeneity, stromal interactions, and a robust immune cell activation across diverse types of IBD. Conclusion These results provide a broad view of the complex cellular mechanisms underlying IBD. Automated high-plex immunophenotyping offers novel insights into immune cell dynamics and tissue organization, potentially guiding precision therapeutic strategies in colonic inflammation. References Selin et al. J Crohns Colitis. 2021 May 7;15(11):1959—1973. Zheng et al. J Inflamm Res. 2025 Jul 9:18:8977-8992. Rivest F et al. Sci Rep. 2023. 13(1):16994. Funding Source n/a Topic Categories Immune Mechanisms of Human Disease (HUM)
Activation-induced marker (AIM) assay to detect Chlamydia trachomatis-specific T cells in highly exposed women 2261007
Abstract Introduction Natural immunity to Chlamydia trachomatis (CT) develops in some women but is weak and requires sensitive assays. Activation-induced marker (AIM) assays detect antigen-specific T cells via surface markers and offer higher sensitivity, reduced cytokine bias, and intact viability compared with intracellular cytokine staining, important for sorting cells for downstream single-cell analyses. Methods PBMCs from 115 women highly exposed to CT (TRAC2) and 12 seronegative controls were stimulated for 24h with chlamydial CPAF or MOMP peptide pools. Cells were stained for viability, CD4, CD8, and six AIMs (CD69, PD-L1, CD25, OX40, CD40L, 4-1BB). AIM+ cells (expressing ≥2 markers) were normalized to a Leukopak control in each batch and background-subtracted using paired media controls. Results TRAC2 CT+ participants had significantly higher AIM+ CPAF- (p = 0.001) and MOMP-specific (p = 0.007) CD4 responses than controls, while CD8 responses were not significantly elevated (p = 0.18 and p = 0.27, respectively). Of CT+ participants, 41% (39/95) and 40% (37/93) had AIM+ CD4 responses to CPAF and MOMP (>2X control mean), respectively, while 20% (19/95) and 15% (14/93) showed corresponding CD8 responses. CD69 and PD-L1 were most commonly co-expressed, but with variable patterns across individuals. Conclusion AIM assays markedly improve detection of CT-specific T cells and support the central role of CD4 T cells in responses to CT infection. This platform will enable sorting and multi-omic profiling of CT-specific T cells across blood, cervical, and endometrial TRAC2 tissues by Ab-, mRNA-, and TCR-seq. AIM+ PBMCs will define individualized CT-specific TCR repertoires to match clonotypes in fragile mucosal samples without ex vivo stimulation. We will assess whether CT-specific cells are skewed toward distinct T-helper phenotypes or transcriptional profiles, whether they differ by antigen (CPAF vs MOMP), and which are linked to reduced recurrent or ascending infection. Funding Source T32 AI007001, R01 AI170959 Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)
T-T cell crosstalk: Insights into mechanisms and disease development 2254071
Abstract Introduction Intercellular signaling is essential for immune coordination and cellular identity. While interactions between innate and adaptive immune cells are established, the mechanisms driving direct T—T cell communication are poorly understood. We hypothesized that CD4 memory T cells (CD4 Tmem) can directly interact with naïve CD8 T cells (CD8 TN) to influence their activation state, and we tested this both ex vivo and in vivo. Methods We identified the phenotype by performing ex vivo co-culture of human CD4 memory T cells (CD4 Tmem) with naïve CD8 T cells (CD8 TN) and assessing surface marker expression (CD45RO, CCR7). To test in vivo relevance, OVA-specific CD8 TN (OT-I) were co-adoptively transferred with OVA-expressing CD4 Tmem into wild-type B6 mice, and activation was measured by surface marker expression. Mechanistic signaling was evaluated using both CD8 TN and CD4 Tmem cells from NR4A-GFP reporter mice, where GFP expression serves as a reporter of NR4A activity downstream of TCR signaling. To assess disease relevance, we developed an ex vivo model using ApoE⁻/⁻ aorta co-cultured with T cells. Results Ex vivo, CD4 Tmem induced an activated-memory phenotype in CD8 TN. In vivo, co-transferred OT-I cells showed activation and differentiation, whereas OT-I cells injected alone did not. CD8 TN co-cultured with activated CD4 Tmem upregulated CD44 and NR4A-GFP, indicating TCR engagement. In the atherosclerosis model, co-cultured CD8 TN and CD4 Tmem cells induced Annexin V+ apoptosis in CD45⁻ aortic cells, whereas CD8 TN alone did not. Conclusion These results highlight a CD4-driven mechanism that activates naïve CD8 T cells and promotes vascular pathology. Funding Source Augusta University Start-up Funds Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)
Skull bone marrow is an early source of antiviral CD8 T cell priming and expansion during neurotropic virus infection 2241010
Abstract Introduction The anatomical locations of generating adaptive immune responses to neurotropic viruses are still being investigated. Skull bone marrow (BM) has recently been identified as a pivotal immune site that can sense and respond to a local insult in the brain. Yet, there is a lack of understanding of the role of skull BM during viral encephalitis. We hypothesized that the skull BM is an important niche for antiviral CD8 T cells priming, expansion, and maintenance during Theiler’s murine encephalomyelitis virus infection. Methods Using peptide:MHC tetramer analysis, we determined that antiviral CD8 T cells are primed in the skull BM in early infection, at time points comparable to cervical lymph nodes. Employing MHC class I conditional knockout mice generated by our research group, we inactivated H-2Kb and H-2Db class I molecules on discrete antigen-presenting cell types. Results We determined using these transgenic mice that viral antigen presentation on dendritic cells (DCs) and endothelial cells is essential for the generation of virus-specific T cells in skull BM during acute viral encephalitis. However, antigen presentation by DCs is not required for long-lived antiviral CD8 T cells that reside in skull BM. Moreover, the use of FTY720 to block lymphocyte migration diminished skull antiviral CD8 T cell numbers, suggesting that peripheral trafficking is required for generating skull antiviral CD8 T cells. Conclusion We present compelling evidence for the importance of skull BM as a responsive site that is highly suitable for rapid generation of antiviral CD8 T cells during acute viral encephalitis. Skull BM also serves as a reservoir for memory T cells, long after the resolution of a neurotropic virus infection. This work sheds light on the skull as a potential target for further modulation of the immune response to viral infections in the brain. Funding Source NIH grants R01 NS5103212 and RF1 NS122174 Topic Categories Viral Immunology (VIR)
Pulse Qualification Technology 2306501
Abstract Introduction In a typical flow cytometer, the signal generated by a single cell crossing the focused light beam produces an electrical voltage pulse. Three meta-parameters–height (H), area (A), and width (W) of the pulse–are traditionally used to characterize cells across scatter and fluorescence channels. However, cell clumping and closely spaced events can produce overlapping pulses, leading to coincidence effects that compromise data accuracy Methods Standard doublet discrimination using area-versus-height plots often relies on subjective user judgment, underscoring the need for more robust and objective gating strategies. Results Standard doublet discrimination using area-versus-height plots often relies on subjective user judgment, underscoring the need for more robust and objective gating strategies. Conclusion Results are shown using biological samples and NIST-traceable calibration beads. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)
Nutritional Regulation of PKCζ: Omega-3 Fatty Acids Shape Neonatal T Cell Development 2255253
Abstract Introduction Infants born to mothers with a family history of allergy often display transiently low cord blood T cell (CBTC) protein kinase C ζ (PKCζ) levels, which are associated with an increased allergy risk. Our previous studies have shown that PKCζ is a key regulator of neonatal T cell development, promoting a Th1 cytokine bias. Maternal supplementation with ω-3 polyunsaturated fatty acids (PUFAs) during pregnancy enhanced neonatal PKCζ expression and protected against allergic sensitisation. The purpose of our investigation was to discover whether PKCζ is both a biomarker of allergy risk and a target for allergy prevention. Methods CBTC and a surrogate CBTC culture maturation model was used to examine the levels of PKCζ and cytokines produced, employing monoclonal antibodies and flow cytometry. The modulatory effects of ω-3 PUFAs was assessed by adding the fatty acids directly to the CBTC. Similarly, when examining the effects of PUFA metabolic products. The role of the fatty acid metabolism was examined by using inhibitors of the lipoxygenase and cyclooxygenase pathways. Results The data demonstrate that ω-3 PUFA treatment caused an increase in PKCζ expressed by the CBTC. Interestingly we found that while some PUFAs caused an increase, others not only failed to increase the PKCζ levels but induced a decrease in this expression. The increase in PKCζ in CBTC induced by ω-3 DHA was reflected in a transition of cells maturing away from a Th2 allergy-promoting cytokine propensity (i.e. Th1 cytokine profile predominance). Its metabolism through the lipoxygenase pathway was import for this effect and it was related to the generation of resolving-D1. Conclusion The findings identify PKCζ as a target in newborn T cell to prevent development towards a Th2-cytokine propensity and potentially allergic sensitisation. These expand the ‘hygiene hypothesis’ to a nutrient environment that regulates PKCζ expression. Funding Source Adelaide Scholarships International (ASI, 2016-2020), University of Adelaide; Invest for Success grant, Robinson Research Institute, The University of Adelaide. Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)
Inorganic Ionic Polymerization‐Anchored Polymer Networks for Ultrastrong and Ultratough Eutectogels
ABSTRACT Eutectogels hold considerable promise for applications in flexible electronics, soft robotics, and protective systems owing to their combination of the environmental stability of deep eutectic solvents and the mechanical robustness of polymer networks. However, predominantly weak intermolecular interactions among polymer chains within eutectogels restrict their mechanical performance, making it challenging to simultaneously achieve high strength and high toughness. Herein, we propose an inorganic ionic polymerization‐anchored polymer network (IIP‐APN) strategy to construct ultrastrong and ultratough eutectogels. Specifically, calcium phosphate oligomers (CPO) are incorporated into the polyvinyl alcohol (PVA) chain network to serve as nanoanchors. Through inorganic ionic polymerization, hydroxyapatite nanorivets are generated to anchor PVA chains, forming a hierarchically integrated organic–inorganic composite network. This unique riveting network structure imparts record‐breaking ultrahigh toughness (696.4 ± 119.1 MJ m − 3 ) and high strength (58.02 ± 3.87 MPa) to the resulting PVA/CPO eutectogels, significantly surpassing existing gel materials. Moreover, the PVA/CPO eutectogels demonstrate excellent energy absorption and dissipation, outstanding fatigue resistance over 8000 cycles, and the capacity for damage repair via secondary inorganic ionic polymerization. Consequently, PVA/CPO eutectogels exhibit significant potential for technological applications. The proposed IIP‐APN strategy provides a powerful platform for the design and development of high‐performance gel materials.
IFNAR and STING signaling restrict proliferation of hapten- primed CD8 T cells producing IFNγ in response to skin sensitization 2255903
Abstract Introduction Contact hypersensitivity (CHS) is a CD8 T cell-mediated response to hapten skin sensitization and challenge. While sensitization of wild-type (WT) mice induces hapten-reactive CD8 T cells that produce the effector cytokines IFNγ and IL-17 required to mediate CHS, the role of type I IFN in CHS is unclear. This study investigated a putative role for type I IFN receptor (IFNAR) signaling during hapten-specific CD8 T cell priming. Methods DNFB sensitization Results Dinitroflourobenzene (DNFB) sensitized IFNAR-/- mice had 2-3 fold more hapten-primed CD8+ T cells producing IFNγ but not IL-17, vs. WT mice in the skin draining lymph nodes and elevated frequencies of CD11ahiCD44hiCD8+ cells proliferating and expressing CXCR3 on day +5 post-sensitization. To begin to test if the IFNAR pathway is activated through cGAS binding, STING mice were sensitized. DNFB sensitized Sting-/- mice had increased numbers of hapten-primed CD8+ T cells producing IFNγ in the skin draining lymph nodes as well as higher frequencies of proliferating CD11ahiCD44hiCD8+ cells that express CXCR3. Numbers of hapten-primed CD8+ T cells producing IFNγ in the skin draining lymph nodes of sensitized Sting-/- mice remained significantly higher than WT at day +8 post sensitization despite the presence of CD4+CD25+ cells. Conclusion These results suggest IFNAR signaling via Sting is required to regulate the magnitude and duration of CHS responses through limitation of effector IFNγ producing CD8 T cell expansion. Funding Source n/a Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)
Natural Antibody Recognition of DAMPs Is Required for Inflammasome-Driven Anti-Tumor Immunity 2257202
Abstract Introduction Damage-associated molecular patterns (DAMPs) released from malignant cells are known to induce immune responses. Natural Antibodies (Nabs), primarily produced by B1 B cells are known to bind DAMPs and clear out excessive DAMPs associated with cell death, preventing autoimmunity. However, the mechanisms linking NAb mediated DAMP recognition to anti-tumor immunity remain unclear. Methods The role of natural antibodies (NAbs) in DAMP-mediated immunity was investigated using transgenic mice deficient in B cells (CD19creDTA) and NAbs (IgHEL MD4). DAMPs including Uric acid, Calreticulin, and HMGB1 were tested for their ability to elicit immune responses against neoantigen-expressing cells. Additional knockout mouse models were used to define signaling pathways triggered by DAMP-NeoAg-NAb complex that promote cytotoxic T lymphocyte (CTL) activation. Results In the absence of NAbs, DAMPs failed to induce cytolytic activity or generate neoantigen-specific CTLs, indicating that the circulating NAb repertoire alerts the immune system to the presence of transformed cells. Moreover, B16F10 melanoma models confirmed that NAbs were required to license DAMP-mediated anti-tumor immunity; without them, a significantly greater tumor burden was observed. However, immunization with DAMPs and B16 peptides and restoration of the NAb repertoire resulted in reduced tumor burden. Conclusion Natural antibodies are essential for DAMP-induced type 1 immunity, enabling early recognition and elimination of transformed or precancerous cells. Funding Source National Institutes of Health (NIH) grants: NIH grants R35 HL155458 (C.V.J.) National Cancer Institute Cancer Center Support Grant 5P30CA023108 (F.W.K.) NIH NIGMS P20GM130454 (F.W.K.) NIH S10 1S10OD030242, (F.W.K.) Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)