bluuebunny/crossref_metadata_2025_split huggingface dataset:
Loss of isoprenylation significantly reduces IgE mediated mast cell activation
Authors: Dailey Jordan , Ballance William , Kee Sydney Ann , Kotha Aditya , Sebti Said , Ryan John J 
Published: January 2023

Abstract
Allergic disease is the 6th leading cause of chronic illness in the US and accounts for billions of dollars in healthcare costs annually. Mast cells are tissue resident innate immune cells linked to allergic disease and 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. Previous findings showed repurposing of statin drugs, such as Fluvastatin, reduced allergic symptoms in vitro and in vivo. We found that Fluvastatin suppresses IgE-mediated mast cell activation by inhibiting isoprenylation of proteins. However, statin effects were dependent on genetic background. To avert genetic background dependency, we tested the hypothesis that a dual farnesyl and geranylgeranyl transferase inhibitor (FGTI 2734) would suppress allergic disease withoutgenetic background limitations. We show that FGTI 2734 reduced IgE-mediated degranulation and cytokine production by bone marrow derived mast cells from multiple genetic backgrounds. Similarly, FGTI 2734 also suppressed IgE-mediated anaphylaxis on multiple genetic backgrounds. These in vivo effects extended to an allergic asthma model, in which FGTI 2734 inhibited pulmonary inflammation. Finally, we show that FGTI-2734 can significantly reduce IgE-induced cytokine production from human skin mast cells. These data show that dual farnesyl and geranylgeranyl transferase inhibitors may be an effective therapeutic treatment for mast cell-associated diseases.
Supported by NIH (1RO1AI101153)
The exon-skipping oligonucleotide, KitStop, depletes tissue-resident mast cells in vivo to ameliorate anaphylaxis
Authors: Hedgespeth Barry , Snider Douglas , Bitting Katie , Arthur Greer , Smith Cierra , Cruse Glenn 
Published: January 2023

Abstract
Anaphylaxis represents the most extreme, life-threatening form of allergic disease and is often an emergency requiring immediate intervention. It occurs following cross-linking of the high-affinity IgE receptor (FcɛRI) on the surface of mast cells by allergen-IgE complexes; however, non-IgE-mediated and idiopathic mast cell activation also contribute. Causes include environmental allergens, foods, venoms, and medications. In addition, some people with mastocytosis have repeated anaphylactic episodes during normal daily activities with no exposure to these triggers. While acute therapy consists primarily of epinephrine and supportive care, chronic therapy relies heavily on desensitization to the inciting allergen—a time-consuming and sometimes unsuccessful process. Desensitization also requires identification of the trigger which is not always possible. Here, we report the use of an exon-skipping oligonucleotide, previously termed KitStop, to safely reduce the severity of anaphylaxis via mast cell depopulation in tissues. KitStop administration results in the integration of a premature stop codon within the mRNA transcript of the Kit receptor—a receptor tyrosine kinase found primarily on mast cells and whose gain-of-function mutation can result in systemic mastocytosis. After either local or systemic KitStop treatment, mice had significantly reduced mast cell numbers in the skin, peritoneum, and lungs. In addition, KitStop-treated mice had a significantly diminished anaphylactic response in a model of passive systemic anaphylaxis. These data demonstrate that KitStop has the potential to serve as a powerful treatment option for patients that experience anaphylaxis, including idiopathic anaphylaxis.
Research was supported by the National Institutes of Health (NIH), National Institute of Allergy and Infectious Diseases (NIAID), Award Number R01AI143985 (G.C.), Department of Molecular Biomedical Sciences, College of Veterinary Medicine, start-up funds (G.C.), NCSU Center for Human Health and the Environment NIH P30ES025128 (G.C.), Institutional National Research Service Award from the Office of the Director of National Institutes of Health, NIH T32OD011130 (B.H., D.S.).
Corticosteroids treatment alters lung and gut microbiome communities that correlates to increased pathologic immune responses
Authors: Asai Nobuhiro , Fonseca Wendy , Yagi Kazuma , Ethridge Alexander D , Morris Susan H , Rasky Andrew J , Falkowski Nicole R , Huang Yvonne J , Huffnagle Gary B , Lukacs Nicholas W 
Published: January 2023

Abstract
Inhaled corticosteroid (ICS) are recommended as first-line controller medications for persistent asthma. Microbial interaction among the gut-lung axis has been suggested. The impact of this crosstalk can modify homeostasis and disease development by altering the host’s immune responses and metabolic status. In this work, we investigated the effect of ICS on microbiome during chronic allergic pulmonary disease. Here we demonstrated that the alternations of lung and gut microbiome can be linked to higher immunological response in mice treated with ICS with Respiratory syncytial virus (RSV) infection. We observed that ICS use during chronic allergy protected the mice, as expected, from lung inflammation and mucus deposition. However, the lung and gut microbiome also presented significant alterations in both naïve mice treated with ICS alone or in ICS asthmatic mice. Microbiome analysis revealed that the principal component analysis of lung and cecum samples in the ICS groups are significantly different from other groups suggesting that ICS impacts the lung and gut microbiome directly with specific organisms altered. When we RSV infected naïve mice treated with ICS, compared to naïve without ICS we observed significant worse pathologic responses. qPCR analysis of Th2-cytokines and of mucus gene showed significantly higher expression in naïve mice treated with ICS than control treated RSV infected mice. Altogether, alternations of lung and gut microbiome can be associated with treatment with ICS with higher immunological response in naïve mice with ICS after RSV infection. Our future studies will focus on how the changed microbiome modifies the subsequent immune responses by focusing on local lung and systemic metabolite changes.
Interorgan transfer of intact micron-sized lipid droplets to macrophages during the Drosophila immune response
Authors: Brennan Catherine A , Kaur Ishneet , Myers Amber , Ogundipe Eniola 
Published: January 2023

Abstract
Macrophages are white blood cells of the immune system in both mammals and Drosophila. In addition to their roles in engulfing and killing microbial invaders by phagocytosis, these cells play important roles in embryonic development, tissue repair, metabolism, and maintenance of homeostasis. Under certain inflammatory conditions, mammalian macrophages accumulate lipids in the form of lipid droplets, including in infection, atherosclerosis, and obesity; however, neither the mechanisms nor purpose of this accumulation are well-understood. We have found that fly macrophages also accumulate lipid droplets, with astonishing speed following bacterial infection: filling up to 30% of cell volume within 3 hrs of infection. We have discovered a completely novel mechanism for this inflammatory lipid accumulation: instead of synthesizing lipids from fatty acids or glucose taken up from the blood, fly macrophages obtain intact, micron-sized lipid droplets from the adipose tissue, through a phagocytosis-like mechanism we term “phagoliposis”. By genetically blocking the metabolism of these lipid droplets, we have discovered some important inflammatory mechanisms fueled by these lipids.
Supported by grants from NIH (SC2AI133653)
Batf3-dependent regulation of the antifungal inflammatory response after pulmonary Aspergillus fumigatus challenge
Authors: Rapp Alexander , Obar Joshua J 
Published: January 2023

Abstract
Aspergillus fumigatus is a ubiquitous fungus that causes invasive aspergillosis in immunocompromised individuals. The innate immune system is critical in the maintenance of host resistance against A. fumigatus. While it is well established that monocyte-derived dendritic cells (moDC) are critical in maintaining host resistance against A. fumigatus, the role of other dendritic cells populations has not been thoroughly explored. Here we demonstrate that Batf3-dependent cells, likely classical type 1 dendritic cells (cDC1s), are critical in preventing invasive aspergillosis. Surprisingly, Batf3(−/−) mice display an elevated inflammatory response against A. fumigatus, particularly IL-17A/F. We show that elevated concentrations of IL-17A/F in the lungs of Batf3(−/−) mice following A. fumigatus infection perpetuates the secretion of various inflammatory lung cytokines in vivo. Neutralization of both IL-17A and IL-17F in Batf3(−/−) mice prevent excessive cytokine secretion and mortality due to A. fumigatus challenge of Batf3(−/−) mice. Following A. fumigatus challenge regulatory T cells are known to limit inflammation. Our preliminary antibody-mediated depletion studies demonstrate that CD4+ CD25+ T cells may be involved in preventing excessive IL-17A/F release in immunocompetent mice. In this study we have identified a critical role for Batf3-dependent cells in limiting A. fumigatus induced inflammatory and mortality due to IL-17 driven lung pathology.
Supported by NIH: [R01 AI139133-01A1]
Skin resident macrophage-derived microRNA21 drives susceptibility to Staphylococcus-aureus infection.
Authors: Salina Ana , Medeiros Alexandra Ivo , Serezani C Henrique 
Published: January 2023

Abstract
Staphylococcus aureus skin infection is orchestrated by the actions of resident and recruited immune cells that lead to abscess formation and bacteria contained. Modulation of the immune response can be achieved by microRNAs. MicroRNA21 (miR21) is a homeostatic regulator of macrophage polarization and miR21 could modulate the expression of either pro- or anti-inflammatory mediators. We are hypothesizing that miR21 is a central node between the regulation of the inflammatory response and antimicrobial effectors functions in the skin. Our data show that miR21 increases as early as 12h, peaks at 24h and 48h. The depletion of skin resident macrophages, but not recruited neutrophils, showed a specific role of skin macrophages in miR21 expression during skin infection. Skin infection in miR21Δmyel mice resulted in reduced lesion size and lower bacterial burden when compared to WT animals. We detected increased IL-1ß and TNF-α and decreased IL-6 and IL-1α levels in the skin of miR21 deficient mice. We also observed increased expression of pro-resolution genes, such as CD36 and collagen3a1 at day 1 after infection. The balance between proinflammatory cytokines, along with the presence of collagen 3 and CD36, resulted in a highly organized abscess with ticker capsule when compared to infected WT mice. Topical treatment with a miR21 antagomir induces reduced lesion size and bacterial burden. Nanostring analysis showed increased Myd88 expression in both naïve and S. aureus infected miR21Δmyel mice and blocking MyD88 actions prevented miR21 antagomir-mediated bacterial clearance in vivo. Our results show blocking miR21 expression leads to a well-controlled inflammatory response necessary to control skin bacterial host defense.
Supported by FAPESP 2018/01622-9, HL-103777, R01HL124159-01
Monocyte/macrophage heterogeneity during skin wound healing in mice
Authors: Pang Jingbo , Maienschein-Cline Mark , Koh Timothy 
Published: January 2023

Abstract
Cells of the monocyte/macrophage (Mo/Ma) lineage orchestrate diverse biological processes required for efficient skin wound healing. Current understanding of Mo/Ma heterogeneity during wound healing has been established by traditional experimental approaches such as flow cytometry, qPCR, and IHC, which indicates a phenotype switch from pro-inflammatory to pro-healing cells as wound healing progresses. However, these phenotypes are limited by the relatively small number of markers assessed. Therefore, our study aimed to generate an unbiased atlas of Mo/Ma phenotypes over the course of skin wound healing by scRNAseq.
Cells were dissociated from excisional skin wounds of normal healing (WT) mice on day 3, 6, and 10 post-injury and live CD11b+Ly6G− cells were isolated by FACS sorting. Using 10× Genomics Chromium system, we pooled data from a total of 2,813 high-quality cells and performed clustering using Seurat which resulted in 8 subpopulations of cells. Cluster dissimilarity and differential expression analysis categorized those clusters into three groups: early stage/pro-inflammatory, antigen-presenting, and late-stage/pro-healing phenotypes that changed over the course of healing. Signature genes and gene ontology analysis of each cluster were used to further characterize the cell populations. Moreover, Ly6C+ vs Ly6C− Mo/Ma were FACS sorted and qPCR was used to validate select marker genes from our scRNAseq clusters. Finally, cell trajectory analysis by pseudotime and RNA velocity indicated the transitions between different Mo/Ma clusters over the course of healing. In summary, our data demonstrated the complexity of Mo/Ma phenotypes and their diverse functions during skin wound healing in mice.
Funded by NIH/NIGMS: R35GM136228
A non-canonical role of caspase-1 in regulating bacterial physiology and antimicrobial resistance
Authors: Akhade Ajay Suresh , Mosquera Gabriela Verdezoto , Arrieta-Ortiz Mario L , Kaur Amardeep , Peterson Eliza J R , Baliga Nitin S , Hughes Kelly T , Subramanian Naeha 
Published: January 2023

Abstract
Caspase-1 is a key effector molecule involved in inflammasome activation and has a well-established role in restricting the growth of intracellular pathogens like Salmonella by triggering a form of cell death called pyroptosis. Here we reveal a non-canonical, cell death independent role for caspase-1 in controlling the transcriptional state and drug resistance of intracellular Salmonella. Using Pathogen-sequencing, a method for sensitive transcriptional profiling of miniscule numbers of intracellular bacteria from infected macrophages, we show that that caspase-1 regulates key processes involved in survival and antimicrobial susceptibility of intracellular Salmonella. Host caspase-1 increased susceptibility of Salmonella to endogenous cationic antimicrobial peptides, as well as to a cationic polypeptide antibiotic used as a last-line drug in Gram-negative bacterial infections. These effects of caspase-1 were independent of its enzymatic activity but dependent on its ability to repress activation of a two-component signal transduction system in intracellular bacteria. These effects were also independent of caspase-11. Our data suggest a “backup” role for caspase-1 in dampening antimicrobial resistance of intracellular Salmonella which evade initial innate immune detection and restriction by caspase-1. These findings also reveal the role of a key innate immune effector in altering pathogen physiology, broadening our view of host-pathogen crosstalk with possible implications for targeting caspase-1 in host-directed therapy to combat antimicrobial resistance.
Supported by grant from NIH (R01 AI155685-A1) and AAI Careers in Immunology Fellowship (2018-2019)
N, N-Dimethylacetamide (DMA) improves survival in a lipopolysaccharide (LPS)-induced endotoxemia model
Authors: Xiao Zhihui , Reznik Sandra 
Published: January 2023

Abstract
Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. In cases of severe sepsis, the risk of mortality is 40% to 60%. Proinflammatory cytokine levels are elevated in patients causing capillary leakage, coagulation, and organ damage. N, N-dimethylacetamide (DMA) is a widely used drug excipient, which was found to possess anti-inflammatory properties. In this project, we studied the potential effects of DMA in improving sepsis survival and organ damage and in regulating the immune system.
To study the potential therapeutic effect of DMA, we utilized an LPS-induced endotoxemia murine model to simulate sepsis-like symptoms. DMA improved 72-hour survival from 17% to 83% in severe endotoxemia. Improved mobility and HR were also observed in DMA treated LPS groups as compared to the LPS only groups. To explore the mechanism of DMA’s ability to improve endotoxemia outcomes, primary murine neutrophils were used. DMA reduced mTNFa levels in primary neutrophils treated with LPS for 24 and 48 hours at 10 mM, the same concentration previously reported to attenuate cytokine secretion in vitro. However, mIL-6 levels were not affected. To further study the effect of DMA on macrophages in vitro, differentiation of human monocyte U937 was induced by PMA. In line with our in vivo and ex vivo studies, U937-macrophages released hIL-6 and hTNFa in response to LPS stimulation while DMA significantly reduced cytokine release when cells were treated with DMA and LPS concomitantly. Additionally, cells co-treated with DMA and PMA during differentiation released less cytokines when compared to the corresponding group treated with PMA only.
Interferon-induced transmembrane protein 3 (IFITM3) limits lethality of SARS-CoV-2 in mice
Authors: Yount Jacob S , Zani Ashley , Kenney Adam D 
Published: January 2023

Abstract
Interferon-induced transmembrane protein 3 (IFITM3) is a host antiviral protein that alters cell membranes to block fusion of viruses. Published reports have identified conflicting pro- and anti-viral effects of IFITM3 on SARS-CoV-2 in cultured cells, and its impact on viral pathology in vivo remains unclear. Here, we show that IFITM3 knockout (KO) mice infected with mouse-adapted SARS-CoV-2 experienced extreme weight loss and lethality, while wild type (WT) mice lost only 10% of their body weight and recovered. KO mice had higher lung viral titers and increases in lung inflammatory cytokine levels, CD45-positive immune cell infiltration, and histopathology, compared to WT mice. Mechanistically, we observed disseminated viral antigen staining throughout the lung tissue and pulmonary vasculature in KO mice, while staining was observed in confined regions in WT lungs. Global transcriptomic analysis of infected lungs identified upregulation of gene signatures associated with interferons, inflammation, and angiogenesis in KO versus WT animals, highlighting changes in lung gene expression programs that precede severe lung pathology and fatality. Corroborating the protective effect of IFITM3 in vivo, K18-hACE2/IFITM3 KO mice infected with non-adapted SARS-CoV-2 showed enhanced, rapid weight loss and early death compared to control mice. Increased heart infection was observed in both mouse models in the absence of IFITM3, indicating that IFITM3 constrains extrapulmonary dissemination of SARS-CoV-2. Our results establish IFITM3 KO mice as a new animal model for studying severe SARS-CoV-2 infection, and overall demonstrate that IFITM3 is protective in SARS-CoV-2 infections of mice.
Supported by grants from the NIH (R01 AI130110, R21 AI151230, R01 HL154001, R21 AI142256, and U54 CA260582)
Role of c-Rel O-GlcNAcylation in regulatory B cells
Authors: Liu Angela R , Ramakrishnan Parameswaran 
Published: January 2023

Abstract
O-GlcNAcylation is a dynamic posttranslational modification, characterized by the addition of an O-linked-β-N-acetylglucosamine to a serine, threonine or tyrosine leading to the fine-tuning of various protein functions. Previously, we have shown that hyperglycemia induces O-GlcNAcylation of the nuclear factor kappa B (NF-κB) transcription factor c-Rel subunit at serine 350. We have found that c-Rel O-GlcNAcylation enhances the transcription of c-Rel dependent proinflammatory cytokines in T cells and inversely regulates FOXP3 expression in T regulatory cells. Other studies have shown that enhanced O-GlcNAcylation dysregulates IL-10 signal transduction and results in decreased IL-10 production. Regulatory B cells primarily secrete IL-10 to conduct its immunosuppressive function. Here, we show that c-Rel O-GlcNAcylation regulates c-Rel-dependent transcription in regulatory B cells. LPS, CD40L, and TLR stimulation under hyper O-GlcNAcylation condition differentially regulates the generation of regulatory B cells from primary B cells isolated from WT and c-Rel knockout C57BL/6 and NOD mice. Mutation of the O-GlcNAcylation site in c-Rel (serine 350 to alanine) also showed differential binding to the IL-10 promoter. This study reveals c-Rel O-GlcNAcylation as a novel mechanism regulating transcription in B regulatory cells with implications in autoimmunity.
ARL was supported by the Visual Sciences Training Program grant from the National Institutes of Health (T32 EY007157). PR was supported by supported by the National Institute of Allergy and Infectious Diseases at National Institute of Health grants R01 AI116730 and R21 AI144264, and National Cancer Institute at National Institute of Health grant R21 CA246194.
Fibrinogen Depletion Ameliorates Inflammation and Vision Loss in Mouse Models of Diabetes
Authors: Cardona Astrid E , Sarker Borna , Cardona Sandra M , Church Kaira A , Vanegas Difernando , Velazquez Priscila , Rodriguez Derek , Mendiola Andrew , Kern Timothy , Muzzio Isabel , Stephens Robin 
Published: January 2023

Abstract
Microglia-mediated inflammation plays a significant role in neuronal and vascular damage in diabetic retinopathy, but the mechanism linking inflammation, neurodegeneration, and impaired vascular integrity is still unclear. Our previous studies from diabetic mouse models showed accumulation of fibrinogen, at vessel lesions surrounded by perivascular microglial clusters. In this study, we evaluated whether the pathological hallmarks of gliosis and vascular aberrations characterized in diabetic animal models are consistent with those in diabetic human retinas. Postmortem human retinas were analyzed by immunohistochemistry for markers of gliosis, vascular integrity, and fibrinogen deposition. Immunohistochemical and gene expression analyses of human postmortem retinas revealed evidence of an inflammatory microenvironment, with microgliosis and impaired vasculature. To define the therapeutic potential of reducing fibrinogen in DR, the defibrinogenating agent ancrod was administered in a two-hit inflammatory diabetic mouse model, after which retinal pathology and visual acuity were assessed. Histopathological analyses revealed microglial activation, vascular aberrations, and fibrinogen deposition in the diabetic murine retina. Notably, after treatment with ancrod, diabetic mice appeared to improve visual acuity, which was associated with reduced microglia activation and less fibrinogen deposition in the retina. This study shows that fibrinogen-mediated microglial activation, blood-retinal barrier damage, and vision loss, can be ameliorated by reducing fibrinogen levels. Overall, these findings suggest that that fibrinogen contributes to microglia-mediated inflammation in the diabetic retina.
Supported by R01 EY029913