bluuebunny/crossref_metadata_2025_split huggingface dataset:
Coronavirus induces diabetic macrophage-mediated inflammation via IFN-beta regulation of SETDB2
Authors: Melvin William James , Audu Christopher , Joshi Amrita , Davis Frank , Barrett Emily , Mangum Kevin , Obi Andrea , Moore Bethany B , Gallagher Katherine 
Published: February 2023

Abstract
COVID-19 induces a robust inflammatory ‘cytokine storm’ that contributes to an increased morbidity and mortality, particularly in patients with type 2 diabetes (T2D). Macrophages are a key innate immune cell population responsible for the ‘cytokine storm’ that has been shown in T2D to promote excess inflammation in response to infection. Using sera from human patients with SARS-CoV-2 and a murine hepatitis coronavirus (MHV-A59) (an established murine model of SARS), we identified that coronavirus induces an increased Mϕ-mediated inflammatory response due to a coronavirus-induced decrease in the histone methyltransferase, SETDB2. This decrease in SETDB2 upon coronavirus infection results in a decrease of the repressive trimethylation of histone 3 lysine 9 (H3K9me3) at NFkB binding sites on inflammatory gene promoters, increasing inflammation. Mϕs with a myeloid-specific deletion of SETDB2 displayed increased inflammation following coronavirus infection. Further, IFNβ directly regulates SETDB2 in Mϕs via JaK1/STAT3 signaling, as blockade of this pathway altered SETDB2 and the inflammatory response to coronavirus infection. Importantly, we also found that SETDB2 mediates a heightened inflammatory response in diabetic Mϕs in response to coronavirus infection. Treatment of coronavirus-infected diabetic Mϕs with IFNβ reversed the inflammatory cytokine production via upregulation of SETDB2/H3K9me3 on inflammatory gene promoters. Together, these results describe a potential mechanism for the increased Mϕ-mediated ‘cytokine storm’ in patients with T2D in response to COVID-19 and suggest that therapeutic targeting of the IFNβ/SETDB2 axis in T2D patients may decrease pathologic inflammation associated with COVID-19.
Combined blockade of IFN-Type I and TNF alpha receptors provides protection against respiratory syncytial virus (RSV)-induced disease and bronchoconstriction
Authors: Morris Dorothea R , Ansar Maria , Qu Yue , Ivanciuc Teodora , Casola Antonella , Garofalo Roberto P 
Published: February 2023

Abstract
While the anti-viral/protective role of interferon type I (IFN-I) and tumor necrosis factor (TNF)-α in naturally acquired RSV infections remains controversial, human and animal studies suggest the existence of a cross-regulation between these two innate pathways, which may contribute to disease and airway inflammation characteristics of this infection. In this study, we investigated the role of IFN-I:TNF-α axis in a mouse model of RSV infection, which provides robust experimental evidence of clinical disease, airway bronchoconstriction and viral replication, recapitulating aspects of the human infection. BALB/c mice were treated with antibodies targeting the IFN-I receptor (IFNR) and either TNF receptor (TNFR)1 or TNFR2 prior to RSV infection. Significant improvements to body-weight loss, illness score and bronchoconstriction measured by plethysmography were observed in mice treated with either antibody combination vs untreated mice (RSV/IgG control). Additionally, several cytokines were significantly reduced in both RSV-IFNR-TNFR mice including IL-1α (p=0.02), IL-6 (p=0.0002), IL-9 (p=0.003), IL-17 (p=0.01), IFN-γ (p=0.005), Ccl3 (p=0.01), Ccl4 (p=0.0003) and Ccl5 (p=0.02). While lung viral load in RSV-IFNR-TNFR1 mice remained comparable to the RSV control (p=0.30), RSV-IFNR-TNFR2 mice had a significant increase in viral replication (p=0.001). Interestingly, this increase in viral replication was not appreciated in previous works blocking IFNR or TNFR2 alone in a similar experimental RSV model. This data collectively demonstrates a synergism between IFN-I and TNF-α during RSV infection as well as highlights the importance of selective receptor blockade when considering RSV therapeutic development.
Identification of a New Lentivirus Restriction Counteracted by Vpr
Authors: Nicodemos Rodrigo A Matus , Ambrozak David , Ransier Amy , Darko Samuel , Douek Daniel C , Koup Richard A 
Published: February 2023

Abstract
Lentiviruses encode non-structural accessory proteins that function to counteract host restriction factors. The lentivirus protein Vpr is known to block G2 to M transition of the cell cycle and degrades various host DNA repair proteins, including UNG2. The reason why Vpr induces these cellular changes in the infected cell is unknown.
We explored this question by direct infection of primary resting and activated CD4 T cells with a CCR5-tropic replication-competent GFP reporter virus. We measured GFP expression by flow cytometry and virus expression by ELISA. We also performed bulk and scRNAseq of sorted GFP+ cells to measure host mRNAs. Additionally, we used a Uracil-qPCR to quantify provirus uracil versus thymidine incorporation. Lastly, we stimulated infected resting cells with and without Vpr and measured cellular proliferation.
We detected resting GFP+ cells 3 to 4 days after infection. Transcriptome analysis of resting GFP+ versus activated GFP+ revealed a pathway for dNTP production in resting CD4 T cells where deoxyuracil is present instead of thymidine. We confirmed provirus uracil incorporation using the Uracil-qPCR assay. Stimulation of infected resting CD4 T cells showed infected cells are preventing from dividing by Vpr. Lastly, we found replication-competent virus was only produced from the initially infected parent cells instead of the divided daughter cells.
We conclude HIV can directly infect primary resting CD4 T cells. HIV-infected resting CD4 T cells incorporate uracils instead of thymidine. After T cell stimulation, Vpr prevents infected cells from dividing because it counteracts an innate lentivirus restriction mechanism against the integrated provirus through UNG2 recognition of the incorporated uracil.
Sodium pyruvate affects influenza A virus infection in vivo
Authors: Reel Jessica Morgan , Lupfer Christopher R 
Published: February 2023

Abstract
Pyruvate is a key metabolite for energy synthesis. After the production of pyruvate through glycolysis, the molecule then gets shuttled into the mitochondria. Once there, it is modified for use in the TCA cycle. While it is a key metabolite for the cell, it has been shown to have anti-inflammatory properties in vitro. Sodium pyruvate has been shown to have effects on metabolic pathways and their influence on the immune response to diseases. Previously we found that infecting mouse bone marrow derived macrophages (BMDM) with influenza A virus (IAV) and treating those macrophages with sodium pyruvate dampened the immune response compared to untreated controls and other infections. We found that subcutaneous injections in vivo, while increasing activity, did not show noticeable differences between placebo and sodium pyruvate treated groups during IAV infection. However, nebulizing sodium pyruvate significantly improved proportional weight loss compared to saline controls. Sodium pyruvate treated groups were found to lose less weight and consume more chow over the course of infection. Nebulizing C57BL/6J mice with sodium pyruvate decreased viral titer and pro-inflammatory cytokines 7 days post-infection as well. Conclusively, nebulizing sodium pyruvate ameliorates IAV infection in vivo.
TRIMming viral infection: identification and characterization of the novel antiviral TRIM7
Authors: Sullender Meagan E , Pierce Linley , Rodgers Rachel , Schriefer Lawrence , Dunlap Bria F , Doench John G , Orchard Robert C , Baldridge Megan T 
Published: February 2023

Abstract
Noroviruses (NoVs) are the leading cause of acute viral gastroenteritis worldwide, causing up to 700 million infections and 200,000 deaths annually, and treatment is limited to supportive care as there are currently no targeted antivirals nor vaccines against NoVs. Therefore, it is critical to discover novel antivirals and develop targeted interventions in order to improve outcomes.
Here we have used a novel genome-wide CRISPR activation screening approach to identify host genes that can inhibit murine norovirus (MNoV) infection or replication. A substantial number of high scoring gene candidates acted in interferon and immune regulation signaling networks, but several were not previously associated with antiviral activity or immunity. TRIM7, the strongest screen hit, was a little studied E3 ubiquitin ligase associated with metabolism and some cancers, yet its expression served as a powerful antiviral in an MNoV-infected cell. Since our initial screen, TRIM7 has begun to emerge as a strategic player in both positive and negative regulation of the innate immune response during infection.
We have since characterized the antiviral mechanism of action of TRIM7 against MNoV in vitro, and have found TRIM7 to be acting early in the viral life cycle to limit viral protein production. Co-immunoprecipitation and in vitro viral evolution experiments have identified MNoV NS6 (viral protease) and NS7 (RNA-dependent RNA polymerase) as important targets of the antiviral activity of TRIM7, and also point to key residues of interest associated with MNoV resistance to TRIM7. Finally, we have investigated the relevance of TRIM7 during in vivo infection using knock-in mouse lines to determine tissue specificity and potency of the antiviral effect.
Antigen receptor on a new lymphocyte represents a key immune player in autoimmune diseases
Authors: Ahmed Rizwan , Omidian Zahra , Giwa Adebola , Ananth Kusuma , Karakus kagan Ege , Aljack Mahamoud , Majety Neha , Yang Angela , Macdonald Alana , Tyagi Sanjana , Zhang Hao , Rabb Hamid , Jie Chunfa , Donner Thomas , Hamad Abdel 
Published: February 2023

Abstract

PURPOSE:
We have recently identified a previously unknown lymphocyte that is a dual expresser (DE) of productively rearranged and surface-expressed TCRαβ and BCR (surface immunoglobulin, Ig) (Ahmed et al, Cell, 2019: 177:11583). Importantly, a single immunoglobulin heavy-chain, IGHV clonotype (clone-x) predominates DEs that encodes a potent autoantigen (x-autoantigen) in its CDR3 region. The x-autoantigen (as a soluble intact x-mAb) cross-activate autoreactive T cells. The goal of this study is to investigate the properties of x-mAb reactive T cells and examining the mechanisms of how x-mAb recognizes and activates the tolerant autoreactive T cells in autoimmune diseases particularly in T1D.

METHODS:
We used EBV immortalized DE clone as a source of x-mAb and FACS-based protocols to identify x-mAb-responsive autoreactive T cells and their functional properties. ImmunoSEQ assay used to characterize TCR repertoires.

RESULTS:
Preliminary data show that x-mAb potentially binds and activates a subset of autoreactive T cells in T1D compared to HC subjects. Additionally, x-mAb-reactive T cells exhibits an activated and antigen experienced phenotype, including expression of CD45RO, CD44, and CD69. TCRVβ repertoire analysis shows that x-mAb reactive T cells are enriched for public clonally expanded TCRs in T1D patients. Further, x-mAb activates the autoreactive T cell through by crosslinking directly to their T cell receptor (TCR).

CONCLUSIONS:
DE cells in T1D patients secretes a public x-mAb that binds and activate specific subset of autoreactive T cells predominated by few clonotypes that express public TCRs. Our results are revealing previously unknown mechanism that appears to be a play critical role in pathogenesis of T1D.
IL-17 receptor A signaling on B cells promotes B-cell responses to TLR7 and enhances autoreactive germinal center B-cell development
Authors: Liu Shanrun , Sun Chiao-Wang , Lu Changming , Ryan Thomas M , Hsu Hui-Chen , Mountz John D 
Published: February 2023

Abstract
IL-17 receptor A (IL-17RA) expression has been identified in B cells and macrophages. There was low expression of IL-17RA on T cells. Surprisingly, we previously showed that IL-17RA signaling through NF-κB p65/p50 has a complex effect on both B and T-cell localization and germinal center (GC) architecture, making it difficult to dissect the specific effects of IL-17RA on B cells. To isolate the effects of the absence of IL-17RA specifically on B cells, we have generated a B-cell specific IL-17RA knockout (Il17rafloxed/floxed x Cd19.Cre) BXD2 mice. Flow cytometry confirmed the loss of IL-17RA expression in only CD19+ B cells but not CD3+ T cells or CD11b+ macrophages. There was a 2-fold decreased spontaneous GC B cell population in Il17rafloxed/floxed x Cd19.Cre BXD2, compared to Il17rawt/floxed x Cd19.Cre BXD2 mice. Interestingly, the abnormal activation and expansion of T-follicular helper cell (Tfh) phenotype in BXD2-Il17ra−/− mice was abrogated in Il17rafloxed/floxed x Cd19.Cre BXD2 mice. In the BXD2-Il17ra−/− mice, B cells were anergic to TLR7 stimulation and exhibited a strong stimulus-specific transcription repressor p50/p50 homodimer and a lower NF-κB phospho-p65. To demonstrate that this anergy is due specifically to IL-17RA on B cells but not other cells, we have identified that there was a lower TLR7-induced CD69 in vitro in Il17rafloxed/floxed x Cd19.Cre BXD2 mice. Taken together, our results suggest that IL-17RA signaling directly regulates B cell autoimmune phenotype in the BXD2 mice through an enhancement of B-cell responses stimulated by the NF-κB signaling pathway. A deficiency of such signal enforces B cell anergy and prevents autoreactive GC formation.
PhIP-Seq analysis reveals autoantibodies for novel antigens in the mouse model of Coxsackievirus B3 infection
Authors: Rasquinha Mahima T , Lasrado Ninaad , Larman Benjamin H , Reddy Jay 
Published: February 2023

Abstract
Myocarditis is a frequent occurrence in children and adolescents that can lead to dilated cardiomyopathy, and viruses are commonly suspected in those affected. Experimentally, Coxsackievirus B3 (CVB) infection in mice occurs in viral and non-viral phases, but in continuum, and histologically, the disease features have similarities with that of human disease. Mechanistically, by using traditional methods, autoantibodies have been demonstrated for various antigens in post-infectious myocarditis leading to a suggestion that autoimmunity could be an underlying mechanism. However, their role continues to be enigmatic, raising a question that whether autoantibody repertoires are sufficiently characterized. Here, we describe a versatile and high-throughput, phage-immunoprecipitation sequencing platform that permitted us to comprehensively evaluate antibody reactivities in serum samples harvested from mice infected with CVB. By screening the phage-displayed virus library (VirScan), antibody reactivity occurred specifically to CVB, and thus, validating our methodology. Similar analysis with the human library revealed antibody reactivity for antigens conserved in both humans and mice, suggesting a role for them in the viral pathogenesis. Alternatively, detection of these autoantibodies may have a diagnostic value in the evaluation of myocardial damage, including cardiac autoimmunity as might occur in CVB infection.
L-type amino acid transporter 1 is essential for Th2 cell-mediated allergic inflammation
Authors: Kaminuma Osamu , Miura Kento , Yamasaki Norimasa , Ogata Sawako , Saeki Mayumi , Hiroi Takachika , Iwata Yohei , Sugiura Kazumitsu , Mori Akio , Endou Hitoshi , Hayashi Keitaro 
Published: February 2023

Abstract
L-type amino acid transporter 1 (LAT1), a transporter of large neutral amino acids, is rarely expressed in resting T cells, whereas its expression is remarkably upregulated upon activation. We have proven the essential role of T cells in the development of allergic inflammation in various target organs, though the contribution of LAT1 to T cell function in allergic diseases has not been clarified. Here, we investigated the effect of a LAT1 inhibitor, JPH203, on antigen-induced inflammatory responses in the skin, lungs, and nasal mucosa of immunized mice or mice transferred with in vitro-differentiated antigen-specific Th2 cells. Consistent with the augmented expression of LAT1 in skin-accumulated CD4+ T cells of atopic dermatitis patients, antigen-induced skin inflammation with eosinophil infiltration and Th2 cytokine production developed in immunized and Th2 cell-transferred mice was suppressed by JPH203 treatment. Essentially the same efficacy of JPH203 was seen in antigen-induced nasal eosinophilic inflammation accompanied by nasal hyperresponsiveness evoked in both mice. Remarkably, antigen-induced bronchial hyperresponsiveness was suppressed, whereas lung accumulation of eosinophils or antigen-specific T cells was not affected, by JPH203 in Th2 cell-transferred mice. The blockade of LAT1-mediated glycolysis, oxidative phosphorylation, and activating transcription factor 4-regulated amino acid starving responses in cytokine production and proliferation of Th2 was suggested to be involved in the efficacy of JPH203. LAT1 is essential for the pathogenesis of activated Th2 cell-mediated allergic inflammation and would be a promising target for the development of new means to treat allergic diseases.
Phytochemical Effects on FcepsilonRI and MrgprX2-mediated Activation of Human Mast Cells
Authors: Kulka Marianna 
Published: February 2023

Abstract
Phytochemicals and herbal extracts are currently used in food supplements as a means of preventing and treating disease. Although phytochemicals have been studied for their potentially beneficial effects on coronary heart disease and their ability to prevent atherosclerosis, certain cancers and inflammatory disease, very little attention has been focused on the potential health benefits of phytochemicals in atopic disease. Mast cells synthesize and store several proinflammatory mediators and are centrally important in atopic diseases such as asthma. Our team has been characterizing the effect of several phytochemicals (flavonoids and polysaccharides) on mast cell activation and pro-inflammatory mediator production when mast cells were activated via the high affinity Fc epsilon receptor I (FceRI) or mas-related G protein coupled receptor, MrgprX2. We show that quercetin and resveratrol decrease mast cell degranulation of both bone marrow-derived mast cells and the human mast cell line (LAD2) by at least 50% (at 10 ug/mL) when the cells were activated via FceRI with IgE and antigen. However, these flavonoids had no effect on LAD2 cells activated via MrgprX2. Polysaccharides isolated from seaweed inhibited IgE/Ag and MrgprX2 activated degranulation of LAD2 cells (by 50%, p&lt;0.01) with some polysaccharides specific to only MrgprX2 activation. These results suggest that mast cell activation can be differentially modulated by phytochemicals.
Characterization of the mast cell surface proteome leads to the identification of CD98hc as a critical surface molecule for optimal mast cell function
Authors: Piliponsky Adrian Martin , Saha Siddhartha , Samanas Nyssa , Shubin Nicholas J , Niino Kerri , Bhise Gauri , Acharya Manasa , Seo Albert , Camp Nathan , Deutsch Gail , James Richard 
Published: February 2023

Abstract
Mast cells are known for their involvement in many distinct pathological conditions, suggesting that mast cells recognize and respond to various stimuli and thus require a rich cell surface protein repertoire. Mast cell surface proteomes have not been comprehensively characterized.
In this study, we aimed to further characterize the mast cell surface proteome to obtain a better understanding of how mast cells function in health and disease.
We enriched for glycosylated surface proteins expressed in murine bone marrow-derived mast cells (BMCMCs) and identified them using mass spectrometry analysis. This approach resulted in the identification of 1270 proteins in BMCMCs, 403 of which were localized to the plasma membrane. The most common protein classes among plasma membrane proteins are represented by small GTPases, receptors and transporters. Novel surface proteins in mast cells was validated by qPCR and flow cytometry analysis in BMCMCs and peritoneal mast cells (PMCs).
Among the novel surface proteins, we found that CD98 heavy chain (CD98hc) encoded by the Slc3a2 gene, was highly expressed in mast cells. Slc3a2 gene disruption by CRISPR/Cas9 gene editing resulted in a significant reduction in CD98hc expression, mast cell degranulation, adhesion and proliferation.
Our study indicates that we can use glycoprotein enrichment coupled with mass spectrometry to identify novel surface molecules in mast cells. Moreover, we found that CD98hc plays an important role in mast cell function.
IL-33 sensitizes mast cells to ATP-induced cytokine secretion
Authors: Pryor Destiny T. , Straus David 
Published: February 2023

Abstract
Mast cells mediate inflammatory responses that promote hypersensitivity reactions and allergic disease. Mast cell activation can be induced by danger-associated molecular patterns such as IL-33 and ATP released by cell necrosis. IL-33 and ATP are elevated in the asthmatic lung, but how these factors act together is poorly understood. Using mouse bone marrow-derived mast cells, we show that IL-33 greatly sensitizes mast cells to ATP. Short term (30 minute) stimulation with IL-33 followed by its removal by washing, increased subsequent ATP-induced cytokine secretion up to 15-fold. This was consistent for ATP-induced IL-6, TNF, and IL-13. IL-33 did not significantly alter mast cell sensitivity to ATP as judged by IC50 calculations, and had no effect on ATP-induced degranulation. We therefore propose IL-33 enhances ATP-induced transcription factor activation without altering calcium-mediated degranulation. We hypothesize that an IL-33-ATP cascade occurs during allergic inflammation and functions as an amplification loop promoting chronic disease.
Internalization of benzylisoquinoline alkaloids by resting and activated bone marrow-derived mast cells utilizes energy-dependent mechanisms
Authors: Alam Syed Benazir , Kulka Marianna 
Published: February 2023

Abstract
Berberine (BBR), a benzylisoquinoline alkaloid, binds to heparin and targets glycosaminoglycan-rich granules of myeloid-derived mast cells, but, the mechanism of berberine’s internalization is poorly understood. Our data showed that 84±3.3% of highly granulated bone marrow-derived mast cells (BMMC) internalized BBR (10 ug/mL) after 45 hr. Methanol fixation completely blocked this process, suggesting an energy-dependent active transport mechanism, and 3-mtheyladenine (600 uM) inhibited internalization by ~43%, indicating that lysosomal self-degradation via the autophagy pathway was involved. BBR incorporation into lipoplexes did not improve BBR internalization, but, rather decreased it by ~30% (1 ug/mL Lipo/BBR; n=3, p&lt;0.05), suggesting that BBR internalization may utilize clathrin-dependent endosomal endocytosis. Interleukin-3 (IL-3)-mediated activation of BMMC augmented BBR internalization (~54% by 40 ng/mL IL-3) in a concentration-dependent manner, further suggesting that clathrin-mediated endocytosis as well as active cell division, have a profound role in BBR internalization. Altogether, our data suggest that internalization of BBR in resting and IL-3 activated BMMC, utilizes clathrin-dependent pathway. Our findings provide important new mechanistic insights into the internalization of benzylsoquinoline compounds by resting and activated mast cells and open new avenues for designing organelle-targeted nanotherapeutics.