bluuebunny/crossref_metadata_2025_split huggingface dataset:
SG-SP1 suppresses mast cell-mediated allergic inflammation via inhibition of FcɛRI signaling
Authors: KIM Min-Jong , Baek Moon-Chang , Kim Sang-Hyun 
Published: January 2023

Abstract
As the number of allergic disease cases increases, studies to identify new treatments take on new urgency. In previous study, gallic acid (GA), a part of EGCG, a major component of green tea, has shown anti-allergic inflammatory effect. However, GA has low metabolic stability and structure-activity relationship. Therefore, we designed SG-SP1 based on GA. We aimed to assess the effects of SG-SP1 on mast cell-mediated allergic inflammation using various animal and in vitro models. For in vitro experiments, IgE-mediated various mast cells (RBL-2H3 and primary cultured peritoneal and bone marrow-derived mast cells) were used to determine the role of SG-SP1 (0.1–1 nM). Immunoglobulin (Ig) E-induced passive cutaneous anaphylaxis and Ovalbumin-induced systemic anaphylaxis are standard animal models for immediate-type hypersensitivity. For in vitro, SG-SP1 reduced degranulation of mast cells by down-regulating intracellular calcium levels in a concentration-dependent manner. In addition, SG-SP1 decreased expression and secretion of inflammatory cytokines in activated mast cells. This suppressive effect was associated to inhibition of the phosphorylation of Lyn, Syk and Akt, and the nuclear translocation of nuclear factor-κB. Due to the strong inhibitory effect of SG-SP1 on Lyn, the known upstream signaling of FcɛRI-dependent pathway, we confirmed the direct binding of SG-SP1 to FcɛRI, a high affinity IgE receptor by surface plasmon resonance experiment. We determined that SG-SP1 directly interacts with FcɛRI and propose SG-SP1 as a therapeutic candidate for mast cell-mediated allergic inflammatory disorders via inhibition of FcɛRI signaling.
IgE-induced mast cell function is suppressed by antagonizing isoprenylation or Ras activation
Authors: Dailey Jordan , Ballance William , Sebti Said M , Ryan John J 
Published: January 2023

Abstract
Mast cells are tissue resident innate immune cells activated by IgE and other ligands. Upon activation they release histamine, cytokines, chemokines, proteases, and lipid mediators evoking allergic symptoms. New ways of targeting mast cells could greatly benefit allergic disease therapy. We previously found that statin drugs reduced IgE-mediated inflammation in vitro and in vivo by inhibiting production of the isoprenoid lipid geranylgeranyl pyrophosphate. However, this suppression was dependent on genetic background. To overcome this limitation, we tested a dual farnesyl and geranylgeranyl transferase inhibitor (FGTI 2734). Using mouse mast cells, we show that FGTI 2734 reduced IgE-mediated degranulation and cytokine production similar to statin effects. Unlike statins, FGTI 2734 did not show a significant difference in IC50 based on genetic background. Furthermore, FGTI 2734 suppressed IgE-mediated systemic anaphylaxis in vivo. Ras GTPases are important for mast cell function and among the signaling proteins modified by isoprenylation. We show that Pan-Ras inhibitors can mimic statin and FGTI 2734 effects on cytokine production in mouse mast cells. These data indicate that dual farnesyl and geranylgeranyl transferase inhibition may be effective in suppressing IgE-induced inflammation, possibly due by disrupting Ras-mediated signals.
Developing a nonhuman primate basophil activation assay by flow cytometry
Authors: Chu Haiyan , Lucchini Amanda , Frantz Aric 
Published: January 2023

Abstract
The Human Basophil Activation test (BAT) has been validated for many disease conditions, used for the diagnosis of allergy to drugs and foods, and for monitoring the role of basophils in immunotherapy. While several BAT kits are commercially available and widely used for humans, the BAT assay in nonhuman primate (NHP) is scarcely present in the literature. Given that NHPs are used as a nonclinical model in drug development and safety assessment when other animal species cannot be used, we developed a BAT using peripheral blood of cynomolgus macaques (Cyno). For human, the mAb panels 1) CCR3+SSClow,2) CD203c+SSClow, 3) CRTH2+CD3−, and 4) CD66−CD3−CD123+HLA-DR− have been used to identify total basophils. Following stimulation with allergens or a positive control (eg. anti-human FcɛRIα or IgE), the activated basophils can be gated as CD63+ and/or CD203+. For the Cyno BAT development, first, the anti-human mAbs noted above were tested for their cross-reactivity with Cyno. Second, the anti-human FcɛRIα and IgE mAbs were tested for their ability to crosslink and activate the Cyno basophils. Third, the basophil stimulation buffer (BSB) was formulated with calcium and heparin with or without IL-3 for blood collected in EDTA tubes. The FcɛRIα but not the IgE mAb, consistently activated basophils and upregulated CD63 expression. IL-3 added to BSB resulted in high background CD63 expression on unstimulated basophils and therefore was not used in the finalized method. Our data indicates that the only panel with all mAbs cross-reactive to Cyno was 4) CD66−CD3−CD123+HLA−. This panel identified the Cyno basophils that were SSCMed not SSCLow. In summary, a BAT has been successfully developed for use in exploratory NHP studies.
Neural Dysfunction Following Francisella tularensis Infection
Authors: Tena Anahis , Muniz Mireya G Ramos , Spencer Charles T 
Published: January 2023

Abstract
Francisella tularensis is a gram-negative intracellular coccobacillus that is the causative agent of tularemia. Local, acute inflammation is beneficial since it can clear bacterial infections, remove cellular damage and promote the healing of tissue. However, F. tularensis is capable of bypassing the immune system and causing an overactivation of inflammation, known as a “cytokine storm”. This cytokine storm increases host cell damage, nutrient availability and serves an immune distraction allowing the bacterium to replicate. Other infections that cause cytokine storms include ebola, pandemic influenza, hantavirus, and HIV, all BSL3 pathogens.
The routes of transmission for F. tularensis are many and intertwined. Infection could be the result of inhaling air droplets containing infectious bacteria, coming into direct contact with an infected animal or being bitten by an infected arthropod such as a tick, flea or mosquito. Eating and drinking contaminated meat and water can also lead to infection and different disease manifestations occur depending on the route of transmission.
While extensive research has documented infection of the spleen, liver and lung, there hasn’t been any reports of F. tularensis productively infecting the brain, until now. This work focuses on characterizing the effects of F. tularensis infection of the brain. We hypothesize that damage caused by F. tularensis neural infection will have a negative effect on behavior. This work aims to identify these negative effects based on the anatomical location of the bacteria foci, regions containing numerous bacteria. With the use of brain mapping and behavioral assays, we seek to determine the impact of F. tularensis infection on behavior and cognition.
Essential role of CCL2-CCR2 axis in the pathogenesis of chronic irritant contact dermatitis
Authors: Shibuya Rintaro , Kitoh Akihiko , Hanakawa Sho , Ishida Yoshihiro , Kabashima Kenji 
Published: January 2023

Abstract
Chronic irritant contact dermatitis (ICD) including hand eczema is a common skin disorder caused by the repetitive exposure to irritants such as chemical detergents and organic solvents. Products including sodium dodecyl sulfate (SDS), such as soap or shampoo, are the leading cause of occupational ICD. Previous studies have shown that irritants cause epidermal damage which induce the production of proinflammatory mediators, followed by a dermal influx of inflammatory cells. However, the detailed immunological cascades occurring in the chronic ICD remains unclear.
Repeated application of SDS, onto murine ear skin induced ear swelling, accompanied by epidermal hyperplasia and barrier dysfunction. Dermal infiltration of neutrophils, eosinophils and monocytes were remarkable in the SDS-applied skin. Our observation described above was consistent with the features of human chronic ICD. Here, we found that CCR2 deficiency or blockade of CCL2, a major ligand for CCR2, significantly attenuated SDS-induced ear swelling as well as inflammatory cell accumulation. Among the differentially expressed genes, which were up-regulated by SDS application, genes related to cell interaction at the vascular wall were significantly affected by CCR2 deficiency. Further, we investigated whether CCR2 on radiosensitive cells or radioresistant cells is critical. Lethally irradiated CCR2−/− mice transplanted with CCR2+/+ bone marrow cells (BMC) exhibited impaired SDS-induced ICD, while CCR2+/+ mice transplanted with CCR2−/− did not.
Collectively, our data suggest that CCR2 expression by radioresistant cells plays an essential role in the pathogenesis of chronic ICD.
Cytosolic LPS sensing triggers the release of a lectin DAMP that promotes inflammation
Authors: Russo Ashley Jean , Vasudevan Swathy , Menoret Antoine , Duduskar Shivalee , Fettis Margaret , Li Chuan , Huergo Santiago Mendez , Kumari Puja , Chandran Karthik , Vanaja Sivapriya Kailasan , Bauer Michael , Sponholz Christoph , Rabinovich Gabriel , Hudalla Gregory , Vella Anthony , Deshmukh Sachin , Zhou Beiyan , Rathinam Vijay 
Published: January 2023

Abstract
Activation of inflammatory caspases, such as caspase-11 and caspase-4, by lipopolysaccharide (LPS) from Gram-negative bacteria, is a key mechanism of innate immune defense against infections. This cytosolic LPS sensing pathway results in activation of a pore-forming protein, gasdermin D, which mediates an inflammatory form of cell death called pyroptosis. In addition, caspase-11-mediated cytosolic LPS sensing leads to the activation of caspase-1 and the maturation of IL-1b and IL-18. Importantly, cytosolic LPS-induced pyroptosis is associated with the release of intracellular proteins that act as alarmins or damage-associated molecular patterns (DAMPs) to modulate the inflammatory response and play an important role during infectious diseases and sepsis. However, the identity and function of DAMPs released downstream of caspase-11 activation are poorly characterized. Using a proteomic approach involving ProteomeLab’s PF2D fractionation system followed by mass spectrometry, we have discovered that galectin-1, a b-galactoside-binding lectin, is released as a consequence of cytosolic LPS sensing. By utilizing various mutant cell lines and primary cells we show that galectin-1 secretion is dependent on caspase-11 and gasdermin D, but not NLRP3 or caspase-1. Importantly, in vivo studies using galectin-1-deficient mice showed that galectin-1 acts as a DAMP to promote systemic inflammation and plays a detrimental role during septic shock. Additionally, human septic patients were found to have increased levels of galectin-1 in their serum. Taken together, we have characterized the secretion and function of a novel lectin DAMP released upon lytic cell death and provide new insights into outcomes of inflammasome activation.
Innate immunity and bacterial conflict: the deep origins of cGAS-STING signaling and discovery of uncharacterized animal immunity pathways
Authors: Burroughs A Max , Aravind L 
Published: January 2023

Abstract
Just as animal innate immunity pits endogenous response pathways against invading extragenic threats, microbiomes are governed by myriad pathways mediating inter-genomic resource procurement, collectively known as ‘conflict systems.’ Leveraging the continually-increasing amount of genome data, we have probed such systems and in the process uncovered the evolutionary origins of core components of animal immunity. This includes the discovery that cyclic-nucleotide synthesizing enzymes like human cGAS and OAS1 and their receptors like STING were acquired from comparable conflict-triggered nucleotide signaling systems in bacteria. While our analyses point to previously-unknown diversity animal innate immunity, particularly in marine invertebrates, this diversity pales in contrast to bacterial pathways which contain a wealth of completely uncharacterized nucleotide synthetase enzymes, nucleotide receptors, and response effector domains.
Further, studies from our group point to entirely-uncharacterized components of the animal innate immunity response that have also been acquired from bacterial conflict systems. These include the viral RNA-acting human Leng9 phosphoesterase and the human N4BP2L1 nucleotide kinase. These studies also uncover a connection between immunity-driven second messenger nucleotide generation and a previously-unrecognized receptor domain in the human apoptotic factor MAP3K5.
The Rab11-family interacting protein 2 is an important regulator of inflammatory cell death in human macrophages
Authors: Husebye Harald , Gravastrand Caroline , Skjesol Astrid , Yurchenko Mariia , Jakovljev Aleksandra , Afset Jan Egil , Espevik Terje 
Published: January 2023

Abstract
Evidence that membrane trafficking and anchoring at the trans-Golgi network is important for NLRP3 inflammasome activation is now emerging. The Rab11-family interacting protein 2 (FIP2) is a key regulator of intracellular vesicle trafficking and has been shown to bind to a number of phosphatidylinositol phosphate (PtdIns) species, including PtdIns4P that is prevalent in the membranes of the Golgi apparatus.
Here we report a role of FIP2 in NLRP3 inflammasome activation downstream of TLR4. High resolution confocal microscopy data revealed that FIP2 was resident in the Golgi and the trans-Golgi-network. In LPS primed and Nigericin treated cells, FIP2 was frequently observed on specks containing the inflammasome adaptor ASC. FIP2 positive ASC-specks were observed in mouse macrophages, THP-1 monocytes, primary human monocytes and macrophages.
In addition, FIP2 showed a marked co-localisation with Caspase-1 in Golgi. Also, FIP2 co-localised with NLRP3 in additional cellular compartments. Indeed, co-immunoprecipitation studies in HEK293 cells showed that FIP2 could bind both NLRP3 and Caspase-1.
Moreover, FIP2 silenced THP-1 monocytes showed impaired ASC-speck formation and marked reduction in secretion of cleaved IL1-β and Caspase-1. Similar results were found when FIP2 silenced THP-1 cells were LPS primed and treated with virulent clinical isolates of uropathogenic Escherichia coli (UPEC). Together these data show that FIP2 is a regulator of NLRP3 inflammasome assembly and activation.
DDX39A interacts with LGP2 to inhibit RLR responses
Authors: Stone Amy E.L. , Thompson Ahna B , Jamieson Abby W , Feliciano Shelby , Said Ali , Gale Michael J 
Published: January 2023

Abstract
RIG-I-like receptors (RLRs) are DEAD-box helicases and cytoplasmic pathogen recognition receptors that recognize and bind to nonself/viral RNA and trigger innate antiviral immunity. RLRs are essential for the detection of RNA virus infection and include RIG-I, MDA5, and LGP2. LGP2 has been implicated as RIG-I or MDA5 co-factor to regulate their activity. We conducted a yeast 2-hybrid (Y2H) screen (using LGP2 as bait) of a human hepatic cDNA library to identify protein binding partners of LGP2. Identified proteins were validated as LGP2 interactors through overexpression and co-immunoprecipitation assays of LGP2 binding. DDX39A, a DEAD Box helicase protein with known roles in RNA splicing, was identified as a binding partner. DDX39A overexpression was found to inhibit interferon-beta production from RLR-signaling in a cell-based ligand-free model in a dose-dependent manner. Further, in a cell-based interferon beta-promoter-luciferase assay model of Sendai virus (SenV) infection, DDX39A and LGP2 synergistically inhibited promoter activity. Similar results were seen in a cell-based NFkB-luciferase assay of the same system. Endogenous expression of DDX39A was assayed through the Huh7 human hepatoma cell line system with various stimuli including viral infection with SenV and Interferon-beta treatment showing that DDX39A is constitutively expressed in those cells. Structure-function mutants of DDX39A and LGP2 were used to determine the protein domains and functions required for interaction via co-immunoprecipitation assays. Through these studies, we will define the interactions between LGP2 and DDX39A, and reveal the roles these proteins have as co-factors in RLR signaling.
Complement regulators and mechanisms vary by sex in intestinal ischemia reperfusion injury
Authors: Wu Miaomiao , Rowe Jennifer , Fleming Sherry D 
Published: January 2023

Abstract
Complement activation plays a pivotal role in intestinal ischemia reperfusion (I/R)-induced injury, leads to significant morbidity and mortality. The injury includes significant inflammation including complement activation. Although established primarily using a male models, intestinal I/R-related diseases occur in both sexes. However, multiple animal models of ischemia and complement activation demonstrate sexual. Thus, we hypothesized that complement regulation may differ by sex during intestinal I/R altering the timing, quantity or required molecules. Both sexes of C57Bl/6 mice were subjected to Sham treatment or 30 min intestinal ischemia followed by 15 min, 30 min, 60 min, and 120 min reperfusion. We demonstrate that compared to male mice, females sustain attenuated intestinal I/R-induced tissue damage. In response to I/R, males produce PGE2 regulated by COX-2, while females produce LTB4 by A5-LOX. Complement initiation pathways differed with males tending to use classical pathway while females tend to use MBL pathway. Together these data suggest that local control of complement activation and regulation, eicosanoid production and neutrophil infiltration in response to intestinal I/R varies between sexes suggesting that distinct therapeutic intervention may be needed in clinical ischemic diseases.
Single cell RNA sequencing reveals novel inflammatory mechanisms of human acute respiratory distress syndrome
Authors: Rosborough Brian R , Chen Jie , Jiang Yale , Kitsios Georgios , McVerry Bryan , Lee Janet S , Ray Anuradha , Chen Wei , Ray Prabir 
Published: January 2023

Abstract
The acute respiratory distress syndrome (ARDS) is a common etiology of acute respiratory failure that results from overwhelming pulmonary inflammation. Since prior bulk RNA sequence analyses have provided limited insights into ARDS pathogenesis, we used the approach of single cell RNA sequencing (scRNA-seq) to probe ARDS at a higher resolution. Towards this end, scRNA-seq data of peripheral blood mononuclear cells (PBMCs) of patients with pneumonia and sepsis with early ARDS were compared to that of sepsis patients who did not develop ARDS. We detected a previously unrecognized reduced frequency of NK cells in the PBMCs of ARDS patients, which also showed complete loss of IFNGR1 expression. Monocyte clusters from ARDS patients revealed multiple distinguishing characteristics in comparison to monocytes from patients without ARDS. This included a pro-inflammatory signature with up-regulation of multiple type I IFN-induced genes, especially in CD16+ cells. To generate an ARDS risk score, we identified 29 differentially expressed genes in the monocytes of these patients, 17 of which showed a similar profile in cells of patients in independent cohorts. Genes with increased expression in monocytes that may have relevance in ARDS pathogenesis include RAB11A, known to inhibit neutrophil efferocytosis, ATP2B1, a calcium pump that exports Ca2+ implicated in endothelial barrier disruption, and SPARC, associated with processing of pro-collagen to collagen. Together, our findings identify molecules that are likely involved in ARDS pathogenesis that may inform biomarker and therapeutic development.
TGF-beta-mediated lung fibroblast-to-myofibroblast transition exhibits IFN-γ resistance via SHP2/SOCS3 activation
Authors: Chen Chia-Ling , Chang Chun-Jung , Lin Chiou-Feng 
Published: January 2023

Abstract
Idiopathic pulmonary fibrosis is a lethal disease of unknown etiology. Abnormal activation of TGF-β is a common cause in developing fibrotic lesions, where Src homology-2 domain-containing phosphatase 2 (SHP2) plays a critical regulator in fibroblast activation and fibrosis. We found that a marked expression of phospho-SHP2, phospho-STAT3 and the suppressor of cytokine signaling 3 (SOCS3) were detected in bleomycin-induced rat lung fibrosis, which was highly association with pulmonary parenchymal lesions and collagen deposition. Human pulmonary fibroblasts differentiated into myofibroblasts via TGF-β1 treatment were characterized with the increased bundles of F-actin, enhanced expression of α-smooth muscle actin (α-SMA), fibronectin and collagen, which also exhibited the activation of SHP2, SOCS1/3, PIAS1, STAT3, IL-6 and IL-10. An AKT-regulated glycogen synthase kinase (GSK)-3β/SHP2 axis was induced in ROS-dependent regulation during myofibroblast differentiation. The significant retardation of interferon (IFN)-γ signaling in myofibroblasts was revealed by the decrease expression of phospho-STAT1 and IFN-γ-associated genes along with IFN-γ-inducible protein 10 (IP-10) reduction. Treatment of nintedanib (bibf1120) in myofibroblasts effectively inhibits both SHP2 and SOCS3 activation accompanying by a suppression of fibroblast-to-myofibroblast transition (FMT). Moreover, synergistic treatment of nintedanib and IFN-γ markedly ameliorated TGF-β1-mediated FMT. Accordingly, we speculate that the activation of both ROS/AKT/GSK-3β/SHP2 and IL-6/STAT3/SOCS3 axis signaling causes myofibroblasts hyposensitivity to IFN-γ whereby escaping from IFN-γ immune-surveillance during FMT.
PU.1 and IRF8 modulate the activation of NLRP3 inflammasome via regulating the monocyte-specific expression of the NLRP3 and its component genes
Authors: Yashiro Takuya , Yamamoto Machiko , Araumi Sanae , Nishiyama Chiharu 
Published: January 2023

Abstract
NLRP3 inflammasome plays crucial roles in the host defense against pathogens by activating Caspase-1, which catalyzes the processing of immature IL-1β and IL-18 into a biologically active form. NLRP3 expression is restricted in monocytic lineages such as monocytes, macrophages, and dendritic cells, thus its expression must be strictly regulated. However, the regulatory mechanism of lineage-specific transcription of the NLRP3 gene remains largely unknown. In this study, we investigated the molecular mechanisms of cell-type specific NLRP3 expression.
RT-PCR revealed that the NLRP3 gene was dominantly driven from the distal promoter in a human monocytic cell line, THP-1. By reporter assay, we identified several cis-acting elements including an Ets/IRF composite element (EICE), which is frequently associated with monocytic lineage-specific gene regulation, in the distal promoter of the human NLRP3 gene. EMSA and ChIP assay demonstrated that transcription factors, PU.1 and IRF8, both of which are known to play essential roles in the development of monocytic lineage, cooperatively bind to the EICE. The expression of NLRP3 was significantly decreased by the siRNA-mediated knockdown of PU.1 and/or IRF8. In addition, the knockdown of PU.1 and/or IRF8 down-regulated the expression of ASC and Caspase-1, which form inflammasome complex with NLRP3. Furthermore, we found that the silencing of PU.1 and IRF8 dramatically abrogated the processing of pro-IL-1β.
These results indicate that PU.1 and IRF8 are involved in the monocytic lineage-specific NLRP3 expression by binding to the EICE site within the distal promoter.