bluuebunny/crossref_metadata_2025_split huggingface dataset:
Specific expression of a BTB-ZF transcription factor defines innate-like regulatory T cells that are essential for intestinal homeostasis
Authors: Krzyzanowska Agata K , Haynes Rashade A , Kovalovsky Damian , Edelblum Karen L , Corcoran Lynn M , Rabson Arnold B , Denzin Lisa K , Sant'Angelo Derek B 
Published: February 2023

Abstract
Regulatory T cells (Tregs) play a pivotal role in regulating immune responses and are a significant source of the anti-inflammatory cytokine IL-10 in the intestine. Defects in Tregs function lead to the development of systemic and intestinal inflammation. Using a single-cell BTB-ZF transcription factor expression analysis, we identified a distinct subset of Tregs. These cells have an activated phenotype (CD62Llo, CD44hi) and constitutively express the Il10. Moreover, these Tregs accumulate in the intestine and expand in number following DSS induced colitis. The targeted deletion of the defining transcription factor impaired these cells' ability to secrete IL-10, leading to significant disruptions in mucosal architecture and the development of intestinal inflammation following treatment with dextran sodium sulfate (DSS). Thus, the conditional knockout (cKO) mice developed much more severe symptoms leading to the death of 60% of them. In contrast, all wild type mice fully recovered. Notably, adoptive transfer of this Treg subset was sufficient to prevent the death of cKO mice. In comparison, the transfer of cKO Tregs was not able to rescue the mice. Collectively, our data show that we have identified a distinct subset of Tregs and determined a specific member of the BTB-ZF family that controls their function. This rare population of T cells has potent immunosuppressive abilities and, we propose, rather than acutely producing IL-10 in response to activation, these cells maintain intestinal homeostasis through continuous secretion of IL-10. Our findings imply that analogous to NKT cells, some thymic-derived Tregs have innate-like effector functions that are not dependent upon differentiation in the periphery.
Fibroblast-like synoviocytes shape inflammation and wound healing in Lyme arthritis
Authors: Wahhab Amanda , Danner Rebecca , Rouse Joseph , Timmler Sarah , Lochhead Robert B 
Published: February 2023

Abstract
Lyme arthritis (LA) is caused by infection with Borrelia burgdorferi (Bb) and, in many patients, can trigger autoimmune and autoinflammatory pathways. Despite proper treatment, some individuals can develop proliferative synovitis that persists for months or years, a condition known as post-infectious LA. Fibroblast-like synoviocytes (FLS) are the most abundant cell type found in the synovial lesion and are critical in wound healing. The aim of this study is to determine the role of FLS in dysregulated pro-inflammatory and tissue repair pathways in LA. FLS were cultured from joints of C57BL/6 (B6) mice and B6 Il10−/− mice and stimulated with interferongamma (IFNγ) and/or Bb. Cell culture supernatants were used for multiplex cytokine analysis, and RNA was isolated from cells for gene expression analysis. FLS were also used in flow cytometry and color flow imaging to identify immune markers, and in cell migration and proliferation assays as a measure of wound healing. When treated with IFNγ, FLS developed an immune-like phenotype. These immune fibroblasts were hyper-responsive to Bb and produced high levels of inflammatory mediators. FLS were also capable of internalizing Bb and expressing MHC-II molecules, suggesting the ability to present Bb antigen. Moreover, B6 FLS treated with IFNγ and/or Bb displayed an impaired wound healing phenotype, whereas this phenotype was seen intrinsically in both treated and untreated Il10−/− FLS. Under pro-inflammatory conditions seen in joints of patients, FLS likely play a key role in numerous pathogenic pathways, most notably in driving inflammation and blocking appropriate repair of damaged tissue. Further studies will examine the epigenetic pathways in both murine and human synovial tissue.
L-plastin supports macrophage NLRP3 inflammasome activation and pulmonary fibrosis
Authors: Joshi Hemant , Almgren-Bell Alison , Anaya Edgar , Todd Elizabeth M. , Morley Sharon Celeste 
Published: February 2023

Abstract
NLRP3 inflammasome induces IL-1β production and pyroptotic cell death in macrophages to fight against invading pathogens. However, excessive inflammasome activation promotes tissue fibrosis and inflammatory diseases such as acute lung injury, liver injury, and acute respiratory distress syndrome (ARDS). NLRP3 inflammasome activation has been extensively studied, but exact regulatory molecular mechanisms remain unclear.
We have found that the actin-bundling protein L-plastin (LPL) is required for efficient activation of the NLRP3 inflammasome pathway. In ex-vivo stimulation, lung resident alveolar macrophages (AMs) lacking LPL produced less IL-1β and IL-18 upon NLRP3 stimulation. LPL−/− AMs from ASC-citrine reporter mice exhibited reduced ASC speck formation after NLRP3 activation. Similarly, bone-marrow derived macrophages (BMDMs) displayed reduced ASC oligomerization. Downstream of ASC oligomerization, caspase-1 activity and downstream IL-1β and pyroptotic gasdermin-D processing were also reduced in LPL-deficient macrophages. Further, LPL−/− mice displayed reduced peritonitis after monosodium urate (MSU) treatment, an experimental NLRP3-induced inflammatory disease model. Since AMs are crucial for NLRP3-mediated lung fibrosis, we next examined NLRP3-dependent bleomycin-induced lung injury. We found that LPL−/− mice responded mildly to bleomycin and exhibited significantly reduced fibrosis development. Thus, we identify a previously unknown significant function of LPL in regulating NLRP3 inflammasome signaling in macrophages. This finding presents LPL as a potential target for diagnosis and treatment for NLRP3-associated inflammatory disorders such as ARDS, COPD and lung fibrosis.
Distinct lymph node entry efficiencies for CD8+ and CD4+ T cells are eliminated during malnourishment.
Authors: Bowman Lauren A , Goulmamine Syreen , Gibson David , Little Amie , Bupp Melanie Gubbels 
Published: February 2023

Abstract
Previous work suggests that glucocorticoids upregulate expression of CD127, an IL-7 receptor component, on peripheral naïve T cells (NT), even though the total number of peripheral NT declines dramatically during malnourishment. We proposed that CD127 up-regulation contributes to peripheral NT reduction by increasing the scavenge rate of IL-7, providing a mechanism to rapidly adjust the total number of NTs during malnutrition. Each malnourished NT would then receive a larger dose of IL-7 than control NTs. We next wondered if this larger dose might confer additional energy-saving behaviors. NT migration across HEV’s may be a high energy-consuming activity. Thus, we compared lymph node entry rates of adoptively-transferred malnourished and control NT in malnourished and control recipients. Control CD4+ NTs migrated more efficiently than control CD8+ NT, but malnourished CD4+ and CD8+ NT migrated at equivalent rates, regardless of recipient diet. We next analyzed expression of proteins known to be involved in NT migration to uncover the currently unknown mechanisms responsible for these various migration patterns. Flow cytometry analysis revealed malnutrition significantly reduces expression of both components of LFA-1 (CD18 and CD11a), CD49d (a VLA-4 component), and S1PR1 in CD4+ and CD8+ NT. We also compared expression levels of ICAM-1 (CD54), a protein expressed in HEV’s which binds to LFA-1 on migrating NT, in malnourished and control lymph node tissue via confocal microscopy. Improved understanding of altered migration molecule expression during malnourishment should enhance our knowledge of the energy-conserving behavior of NT, as well as uncover strategies to improve vaccination responses in malnourished children.
Podosome dissolution and focal adhesion assembly drive the enhanced migratory capacity of dendritic cells infected by ectromelia virus
Authors: Toka Felix N , Biernacka Zuzanna , Struzik Justyna , Gierynska Malgorzata , Szulc-Dabrowska Lidia 
Published: February 2023

Abstract
Podosomes and focal adhesions (FAs) are both adhesive cytoskeletal structures engaged in cell-matrix adhesion of many different cell types, including dendritic cells (DCs). Despite sharing almost, the same proteins, e.g. vinculin, paxillin, talin and Src family proteins, podosomes and FAs show differences in their architecture, dynamics and function. While highly dynamic podosomes are organized in an actin-rich core surrounded by a ring of adhesive molecules with a perpendicular orientation to the extracellular matrix (ECM), more stable FAs have elongated structure and exhibit tangential orientation to the substrate surface. A switch from podosomes to FAs is often observed during TLR4-mediated DC maturation and is associated with an increased motility of these cells. Ectromelia virus (ECTV) is an orthopoxvirus that can control several different aspects of the cytoskeleton organization and dynamics in DCs and, thus, provides a useful model to study virus-cytoskeleton interactions and cell motility in these specialized immune cells, in which ECTV can productively replicate. Our results show that DCs infected with ECTV completely lose podosomes during early stages of infection, within the viral factory formation phase in the cytoplasm (4 hpi). Instead of podosomes, ECTV-infected DCs develop FAs, which are larger but less numerous than in uninfected control cells. FAs are predominantly observed within long-branched cellular projections formed extensively during later stages of infection (18–24 hpi). Loss of podosomes is associated with enhanced migration of infected DCs in vitro, suggesting that ECTV may potentially exploit DC migration to facilitate dissemination of the virus in vivo.
Inhibition of Epigenetic Reader BET Proteins by Apabetalone Counters Inflammation in Activated Innate Immune Cells from Fabry Disease Patients Receiving ERT
Authors: Fu Li , Wasiak Sylwia , Tsujikawa Laura , Rakai Brooke D. , Stotz Stephanie C. , Wong Norman C.W. , Johansson Jan O. , Sweeney Michael , Mohan Connie M. , Khan Aneal , Kulikowski Ewelina 
Published: February 2023

Abstract
Fabry disease (FD) is a rare genetic disorder with a deficit in the degradation pathway of a glycolipid, globotriaocylceramide (Gb3). Gb3 deposits in various tissues evoke immune-mediated systemic inflammation, driving the progression of FD complications. In particular, cardiac and renal problems, leading causes of mortality in FD patients, are insufficiently managed by enzyme replacement therapy (ERT) in the long term. Here, we examined immune profiles in unstimulated peripheral blood mononuclear cells (PBMCs) from FD patients on ERT relative to untreated patients. The results showed an upregulation of the CCR2/MCP-1 axis, particularly in the presence of renal dysfunction. We also observed an increase of IL12B transcripts in unstimulated PBMCs when tracking the immune status over two years of ERT in patients with early-stage chronic kidney disease (CKD). Interestingly, pro-inflammatory responses in LPS-stimulated PBMCs were countered by apabetalone, a clinical-stage candidate that inhibits Bromodomain and Extra Terminal (BET) proteins, epigenetic readers. Apabetalone treatment dose dependently inhibited MCP-1 and IL-12 production by up to 90%. Apabetalone downregulated the transcription of other pro-inflammatory cytokines including TNF-α, IL-6 and IL-8 by 70%, 48% and 54%, respectively. Reactive oxygen species (ROS) are an indicator of oxidative damage caused by intracellular Gb3 deposits in FD patients. Apabetalone suppressed ROS levels by up to ~70% in LPS-stimulated neutrophils. Hence, apabetalone treatment may reduce pathological inflammation and oxidative stress in FD patients and thus complement ERT to optimize patient outcomes, warranting further investigation of apabetalone as a therapeutic for FD.
A single-cell atlas of the lung mononuclear cell response in Staphylococcus Enterotoxin B-induced Acute Respiratory Distress Syndrome
Authors: Wilson Kiesha Maria , Cannon Alkeiver S , Sultan Muthanna , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: February 2023

Abstract
The emergence of SARS-CoV-2 variants will change the landscape of what we already know about COVID-19, making it important to understand the immunological and molecular mechanism of this illness. The virulent nature of the virus, disease outcomes and lack of suitable facilities for studying virus-infected samples has made the need for small animal models urgent. Staphylococcus enterotoxin B (SEB)-induced Acute Respiratory Distress Syndrome (ARDS) mimics the cytokine storm presented in patients with severe COVID-19 and is a safer alternative than working with infected samples. To identify pathways in pulmonary cells that potentially cause ARDS, we utilized single-cell RNA sequencing (scRNASeq) to profile the whole lung cells of C3H/HeJ mice treated with SEB and healthy controls. While it is well established that SEB induces robust proliferation of CD4+ T cells with the receptor Vβ8.2 by binding of these cells with antigen presenting cells, effects in other cell types is not well described. We found that intranasal administration of SEB induced the proliferation of three subsets of macrophages, neutrophils, and cytotoxic T cells in the lungs. Our data also illustrated a loss of multiple cell types including resident macrophages, multiple subsets of B cells, and club cells. Macrophage subsets displayed increased expression of genes related to interferon signaling and lipid metabolism. This data emphasizes the importance of understanding the role of macrophages in ARDS, and can further provide insights into elucidating the mechanisms not only in SEB-induced ARDS but also in severe COVID-19. This may serve as a model for severe case of the disease. (Supported by NIH P01AT003961, P20GM103641, R01AI129788, R01 ES030144 and R01AI123947)
The chromatin reader SP140 regulates intestinal innate immune responses to microbes
Authors: Fraschilla Isabella Rose , Jeffrey Kate L 
Published: February 2023

Abstract
Genome-wide association studies (GWAS) identified single nucleotide polymorphisms (SNPs) within SP140 that associate with Crohn’s disease (CD) in addition to multiple sclerosis (MS) and chronic lymphocytic leukemia (CLL). SP140 is a transcriptional regulator expressed exclusively in immune cells. It is part of the Speckled Protein family that also contains SP100, SP110, and SP140L and shares high homology with autoimmune regulator (Aire). We have previously shown that SP140 is a critical orchestrator of macrophage identity by occupying and repressing silenced lineage-inappropriate genes. However, the role for SP140 functional domains in regulating macrophages identity and transcriptional responses to microbial sensing remained unknown. Here we show how SP140 influences innate immune responses to microbes. By engineering domain mutants, we investigate how the chromatin-binding domains (SAND, plant homeodomain, and bromodomain) and the caspase activation and recruitment domain (CARD) within SP140 dictate macrophage cytokine production. Furthermore, we determine how SP140 localization to promyelocytic leukemia (PML)-nuclear bodies affects function. Global deficiency of SP140 results in exacerbated colitis in mice, and a loss of SP140 specifically in the hematopoietic compartment results in microbial dysbiosis. Collectively, our work provides functional characterization of the novel chromatin regulator SP140 and how disease variants may contribute to immunological disease.
Regulating the unregulated: what host factors are required for GSDMD pore formation?
Authors: Evavold Charles Lee , Hafner-Bratkovic Iva , Devant Pascal , Kagan Jonathan C. 
Published: February 2023

Abstract
Cleavage of gasdermin D (GSDMD) by inflammatory caspases results in pore formation at the plasma membrane. GSDMD pores are recognized to mediate pyroptotic lysis of the cell or direct translocation of IL-1 family cytokines from the cytosol into the extracellular space. While several regulators of GSDMD cleavage have been identified, positive regulators of pore formation that act after cleavage have occurred are unknown. To identify proteins that promote GSDMD pore formation at the plasma membrane after GSDMD cleavage, we utilized a genome-wide CRISPR/Cas9 screening platform in macrophages. This screen uncovered a novel link between host metabolism and cell death at the level of GSDMD pore oligomerization. Several hits in this screen belong to the Ragulator-Rag complex that serves to activate mTORC1. Our data suggests that this Ragulator-Rag-mTORC1 axis promotes pyroptosis through enforcing functional GSDMD pore formation.
NLRP3 and AIM2 inflammasome-triggered Th17 cell response promotes severe immunopathology in schistosomiasis
Authors: Morales Yoelkys , Miller Emily A , Shecter Ilana , Fitzgerald Katherine A , Golenbock Douglas T , Stadecker Miguel J , Kalantari Parisa 
Published: February 2023

Abstract
Infection with the helminth Schistosoma mansoni can cause exacerbated morbidity and mortality via a pathogenic host CD4 T cell-mediated immune response directed against parasite egg antigens, with T helper (Th) 17 cells playing a major role in the development of increased granulomatous hepatic immunopathology. A role of inflammasomes in intensifying disease has been reported, however, neither the types of inflammasomes involved, nor their impact on the Th17 response are known. Here we show that the observed enhanced egg-induced IL-1β secretion and pyroptotic cell death required both NLRP3 and AIM2 inflammasome activation. Schistosome genomic DNA activated AIM2, whereas reactive oxygen species, as well as potassium efflux, were the major activators of NLRP3. In addition, dual activation of caspase-1 and caspase-8 was required for egg-induced IL17 production. NLRP3 and AIM2 deficiency led to a significant reduction in Th17 responses, suggesting that both play a crucial role in triggering inflammation. We also determined that inflammasome-induced IL-1β promotes the anti-inflammatory Th2 cytokine IL-4. Our findings establish NLRP3 and AIM2 inflammasomes as central inducers of pathogenic Th17 responses in schistosomiasis.
The SARS-CoV-2 proteins NSP1 and NSP13 inhibit interferon activation through distinct mechanisms
Authors: Vazquez Christine , Swanson Sydnie , Negatu Seble , Dittmar Mark , Ramage Holly , Cherry Sara , Jurado Kellie 
Published: February 2023

Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic, infecting over 100 million people and claiming over 2 million lives globally. This drastically highlights the need for understanding the host-virus protein-protein interactions and immune evasion strategies governing SARS-CoV-2 pathogenesis. Our overall aim was to identify antiviral immune targeting during SARS-CoV-2 infection and protein overexpression. To understand if SARS-CoV-2 can block immune signaling, we first examined nuclear translocation of the transcription factors, IRF3 and NF-κB, as nuclear translocation is one step in interferon induction, during SARS-CoV-2 infection of the human airway epithelial cells. We found that in infected cells, there was limited IRF3 and NF-κB nuclear translocation, suggesting SARS-CoV-2 can limit immune activation. Additionally, through promoter luciferase signaling assays, the viral proteins NSP1, the host translation shutoff factor, and NSP13, the RNA helicase, can block signaling to the IFN-β and NF-κB promoters. To further investigate this reduction in promoter signaling, we examined nuclear translocation of NF-κB in the context of NSP1 and NSP13 plasmid overexpression. Surprisingly, we found that while NSP13 reduced NF-κB nuclear translocation, NSP1 was unable to despite its ability to limit promoter signaling. Instead, we found, utilizing Click-it chemistry, that NSP1 reduces host protein synthesis. Last, we found NSP13 targets TBK1 activation by limiting its phosphorylation in a dose-dependent manner. Collectively, our data illustrate that SARS-CoV-2 can block multiple innate immune signaling pathways through distinct mechanisms.
Blockade of IFN-I improves adaptive immune responses and immune coverage after vaccination
Authors: Palacio Nicole M , Dangi Tanushree , Wang Yidan , Chung Young Rock , Sanchez Sarah , Penaloza-MacMaster Pablo 
Published: February 2023

Abstract
IFN-I is a main innate antiviral defense critical for the activation of adaptive immunity. However, we hypothesized that acute viral clearance by IFN-I could limit antigen availability following initial antigen encounter, which could in turn, reduce the priming of the adaptive immune system. To test this hypothesis, we immunized mice with different vaccines or acute viruses together with a low dose of an IFN-I receptor-blocking antibody to induce a short-term abrogation of IFN-I signaling. This resulted in transient increase in antigen levels followed by antigen clearance. Importantly, short-term IFN-I blockade during coronavirus, flavivirus, rhabdovirus, or arenavirus immunization resulted in a potent improvement of adaptive immune responses, which conferred improved protection following future re-infections. Short-term IFN-I modulation also improved immune coverage elicited by viral vaccines or acute viral infections. In particular, administration of an HIV-1 vaccine together with the IFN-I blocker generated highly cross-reactive antibody responses against other HIV-1 clades; and similarly, immunization with a mouse coronavirus (MHV) together with the IFN-I blocker generated more cross-reactive antibody responses against other related coronaviruses, including SARS CoV-2. These data demonstrate a novel mechanism by which innate immunity modulates the magnitude and breadth of adaptive immune responses, providing insights for rational vaccine design.
Titers, prevalence and duration of SARS-CoV-2 antibodies in two waves of a local COVID-19 outbreak
Authors: Hedges Jodi F , Thomspon Macy A. , Snyder Deann T. , Robison Amanda , Taylor Matthew P. , Jutila Mark A 
Published: February 2023

Abstract
Information concerning the development of neutralizing antibodies and their duration will be critical to establishing herd immunity for COVID-19. We sought to evaluate SARS-CoV-2 spike protein receptor binding domain (RBD)-specific antibodies, their duration and capacity for SARS-CoV-2 neutralization in volunteers while the pandemic spread within our community starting in March 2020. Those participants with the highest starting titers had the longest lasting response, up to 9 months post-diagnosis. SARS-CoV-2 neutralization capacity was correlated with anti-RBD antibody levels. The majority of our participants with confirmed COVID-19 diagnosis had very mild or asymptomatic infections. We also detected low and largely non-neutralizing anti-RBD IgG titers in a few participants with no known COVID-19 diagnosis. Finally, we found that antibody responses induced by vaccination were significantly higher than those induced by natural infection. Thus, our study suggests that vaccination is still critical even for those naturally infected or diagnosed with COVID-19.