bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic analysis of Bphse and Shs, novel loci controlling hypersensitivity to histamine in mice
Authors: Raza Abbas , Diehl Sean A. , Finstad Kristiaan H. , Case Laure K. , Krementsov Dimitry N. , Blankenhorn Elizabeth P. , Teuscher Cory 
Published: January 2023

Abstract
Histamine (HA) is a biogenic amine produced by mast cells and basophils that regulates many physiological processes including acute and chronic inflammation, immune regulation, and immediate hypersensitivity reactions. Previously, we mapped Bphs, the autosomal dominant locus in mice controlling hypersensitivity to HA following exposure to either Bordetella pertussis or pertussis toxin (PTX) to chromosome 6 (Chr6). Positional candidate gene cloning identified Bphs as HA H1 receptor (Hrh1/H1R). The Bphs-susceptible H1R allele possesses the Pro263, Val312 and Pro330 haplotype while the Bphs-resistant H1R allele have the Leu263, Met312 and Ser330 haplotype. Unexpectedly, when we tested a panel of inbred wild-derived strains for Bphs, we discovered strains e.g. MOLF/EiJ, that are phenotypically Bphs-susceptible despite carrying the resistant H1R allele. Phylogenetically, these strains are confined to a unique branch of wild-derived strains. This supports the existence of an evolutionary selected mechanism capable of complementing Hrh1resistance. Linkage analysis mapped the locus to Chr6, in linkage disequilibrium (LD) with Hrh1. We have designated this locus Bphs-enhancer (Bphse). Additionally, we identified a second Hrh1-linked gene in aged and CFA immunized SJL/J mice controlling HA hypersensitivity in the absence of Bordetella pertussis/PTX-sensitization. We have designated this spontaneous histamine sensitivity (Shs/Shs). Taken together, these data support the existence of a LD domain harboring multiple genes that control hypersensitivity to HA. The identification of the genes underlying both Bphse and Shs has the potential to significantly extend our understanding of the genetic control of HA signaling.
Saturated fat-induced food allergy: A paradigm for unfolded protein response (UPR)-mediated allergic inflammation
Authors: Samavedam Unni Krishna SRL. , Morris Suzanne C. , Khodoun Marat , Wu David , Potter Crystal , Hogan Simon P. , Finkelman Fred D. 
Published: January 2023

Abstract
Murine food allergy induction and maintenance is promoted by increased epithelial permeability and requires the pro-Th2 cytokines: IL-25, IL-33, and TSLP, but how allergens increase permeability and induce pro-Th2 cytokines is unknown. Saturated fats increase intestinal permeability, promote intestinal epithelial pro-Th2 cytokine expression and induce the UPR in several cell types. Two common food allergens, milk and eggs, contain considerable saturated fat. These observations suggested that saturated fats in food might promote allergy by inducing the UPR, which in turn, induces the pro-Th2 cytokines. To test this, quantitative PCR was used to evaluate the ability of heavy cream and egg yolk plasma (EYP, the fat-containing fraction of egg) to induce UPR-associated and pro-Th2 cytokine gene expression in transformed human intestinal epithelial CACO2 cells. Metformin, 4-PBA and TUDCA were used to block the UPR. EYP and cream, but not protein-rich, lipid-poor egg white, increased CACO2 cell UPR-associated gene expression, followed by increased pro-Th2 cytokine expression. This was blocked by the UPR antagonists and by a lipase antagonist that prevents fat absorption. EYP and cream, but not egg white, also increased epithelial permeability; this was blocked by metformin. In BALB/c mice, food allergy (detected as hypothermia and diarrhea) was induced by skin sensitization with egg white + EYP, followed by oral gavage with egg white + EYP. Metformin and TUDCA each prevented food allergy development and strongly suppressed established food allergy. These observations support our hypothesis and suggest that the saturated fat content of egg and milk contributes to their allergenicity.
A disintegrin and metalloproteinase 17 is required for IL-33 induced ILC2 expansion and cytokine production.
Authors: Lownik Joseph Cornelius , Martin Rebecca , Conrad Daniel 
Published: January 2023

Abstract
Type 2 innate lymphoid cells (ILC2s) rapidly respond to the alarmins IL-25 and IL-33, resulting in the initiation of downstream Th2 responses, particularly in parasitic helminth infections. We show that A disintegrin and metalloprotease (ADAM) 17 is required for IL-33 mediated ILC2 activation. In vivo, mice that lack ADAM17 specifically on ILC2s (ADAM17ILC2−/−) exhibit decreased ILC2 expansion in response to intranasal IL-33 as well as Nippostrongylus brasiliensis (Nb) infection. In both of these models, the few ILC2s present in ADAM17ILC2−/− produce less IL-13 than WT ILC2s. Loss of ILC2 expansion in an Nb infection resulted in decreased numbers of IL-13 producing T cells as well as increased worm burden. However, ADAM17ILC2−/− mice have normal ILC2 numbers in a naïve state, suggesting this defect in ILC2 function is limited to cell activation. In vitro, IL-33 treatment of purified ILC2s in the presence of ADAM17 inhibitors show increased apoptosis and decreased IL-13 production compared to control (no ADAM17 inhibitor) ILC2s. The defect in cytokine production following ADAM17 inhibition was not observed in response to IL-25 stimulation, suggesting this defect is limited to IL-33 stimulation Mechanistically, ADAM17 inhibition in ILC2s specifically causes a defect in IL-33 mediated ERK activation, potentially explaining the reduced survival and IL-13 production following ADAM17 inhibition in these cells. Overall, these studies provide a targetable protease to target for the treatment of ILC2 mediated diseases.
Sphingosine-1-phospate receptor 4 regulates passive systemic anaphylaxis in mice but is dispensable for canonical IgE-mediated responses in mast cells
Authors: Kulinski Joseph M. , Proia Richard L. , Tuymetova Galina , Metcalfe Dean D. , Olivera Ana 
Published: January 2023

Abstract
Mast cells are key players in the development of inflammatory allergic reactions. Cross-linking of the high-affinity receptor for IgE (FcɛRI) on mast cells leads to the generation and secretion of the sphingolipid mediator, sphingosine-1-phosphate (S1P) which can in turn activate its G-protein coupled receptors on mast cells. Previous reports have identified the expression of two of the five receptors for S1P on mast cells, S1P1 and S1P2, with functions in IgE-receptor-mediated chemotaxis and degranulation, respectively. Here, we show that cultured human and mouse mast cells also express abundant message for the S1P4 receptor and demonstrate its presence at the plasma membrane in the human mast cell line LAD2. Genetic deletion of S1pr4 resulted in exacerbation of passive systemic anaphylaxis to IgE/anti-IgE in mice. However, exhaustive characterization of IgE-mediated responses in S1P4-deficient bone marrow and peritoneal mouse mast cells indicated that this receptor is dispensable for normal in vitro mast cell growth and responses to IgE-mediated activation including degranulation, cytokine/chemokine release and chemotaxis. Thus, we provide evidence that S1P4 modulates mast cell-mediated anaphylaxis in vivo in an unexpected manner that does not involve regulation of mast cell responsiveness to IgE stimulation.
RNA Binding Protein HuR is critical for the regulation of IL-2 Expression in CD4+ T Cells
Authors: Taylor Tiffany , Ellis Jason S. , Ridenhour Suzanne , Atasoy Ulus 
Published: January 2023

Abstract
The RBP HuR regulates T helper cell differentiation and gene expression by binding to AU-rich elements in the 3′ UTR of its target mRNA and modulating localization and stability. We hypothesized that HuR plays an important role in the regulation of the IL-2 signaling during T cell differentiation and activation. To test this hypothesis, we used the conditional dLck-Cre ROSA HuRfl/fl knock-out mouse to delete HuR late in T cell development but prior to T cell activation. The OVA challenge model of airway inflammation was used to ascertain the effects of HuR KO in vivo. While only 50% of CD4+ T cells in HuR KO mice lack HuR, in vivo and in vitro Th2 responses were essentially nonexistent and there were only background levels of lung inflammation using an OVA challenge model of airway inflammation. The few lung infiltrating CD4+ T cells present had reductions in IL-2 and CD25 expression. HuR KO T cells also had profound antigen-specific proliferation defects in vitro compared to OVA immunized controls and non-specific CD3/CD28 stimulation only partially restored proliferation defects. Furthermore, there is a decrease in IL-2Rα (CD25) which results in decreased levels of phosphorylated (active) STAT5 protein and Blimp1 mRNA and protein levels. To further assess HuR role in regulating IL-2, we developed the pSTAT5+ dLck-Cre HuRfl/fl mouse model, containing a transgene producing constitutively active STAT5. In these mice, there is significant pSTAT5 protein in activated CD4+ T cells, which should circumvent IL-2 downstream signaling, and correct the observed phenotypes. pSTAT5+ HuR KO T cells also had alterations in their expression of multiple CD3 subunits. We conclude that HuR is essential for regulating IL-2 homeostasis and T cell differentiation.
Manipulation of CXCR4 signaling by human cytomegalovirus (HCMV) interleukin-10
Authors: Spencer Juliet , Tu Carolyn 
Published: January 2023

Abstract
Herpesviruses have developed numerous strategies for manipulating host immune responses to avoid clearance and establish lifelong latent infection. Human cytomegalovirus (HCMV) encodes cmvIL-10, an ortholog of human interleukin-10 that binds to the cellular IL-10 receptor (IL-10R) with high affinity to elicit potent immunosuppressive effects, including inhibition of inflammatory cytokine production. In direct contrast, we found that cmvIL-10 promotes heightened chemokine signaling. CXCR4 is an essential cellular chemokine receptor that guides cell movement toward its natural ligand, CXCL12. We found that monocytes and epithelial cells exhibited significantly increased calcium mobilization and migration to CXCL12 in the presence of cmvIL-10. Treatment with a Stat3 inhibitor or siRNA blocked these effects, indicating that Stat3 activation is required for potentiation of CXCR4 signaling activity by cmvIL-10. Using immunofluorescence microscopy, we found evidence of association between IL-10R and CXCR4 in cell membranes and observed that cmvIL-10 treatment led to dramatic changes in the cellular distribution of Stat3. Since intracellular calcium release is a rapid response, Stat3 is likely acting here via a non-canonical pathway, rather than in its classic role as a transcription factor. These findings demonstrate that cmvIL-10 triggers a complex series of events that augment the signaling activity of a cellular chemokine receptor. Given that monocytes are a primary reservoir of latent virus, manipulation of CXCR4 signaling may enable HCMV to modify cell trafficking patterns to promote virus dissemination.
GILT in thymic epithelial cells preferentially facilitates MHC class II-restricted presentation in generating central T cell tolerance to a self and melanoma antigen
Authors: Meador Lydia , Rausch Matthew P. , Metzger Todd C. , Anderson Mark S. , Hastings Karen Taraszka 
Published: January 2023

Abstract
Central tolerance prevents autoimmunity, but limits T cell responses to tumor antigens that are self antigens. In peripheral APCs, gamma-interferon-inducible lysosomal thiol reductase (GILT) is critical for MHC class II-restricted presentation of disulfide bond-containing proteins, including the self and melanoma antigen tyrosinase-related protein 1 (TRP1). The role of GILT in thymic antigen processing and generation of central tolerance is unknown. We found that GILT enhanced the clonal deletion of TRP1-specific thymocytes. GILT expression was enriched in thymic APCs capable of mediating deletion, namely medullary thymic epithelial cells (mTECs) and dendritic cells. TRP1 expression was restricted to mTECs. GILT facilitated MHC class II-restricted presentation of endogenous TRP1 by pooled thymic APCs. Using bone marrow chimeras, GILT expression in thymic epithelial cells (TECs), but not hematopoietic cells, was sufficient for deletion of TRP1-specific thymocytes. Chimeras with GILT expression in TECs also had an increased frequency of TRP1-specific regulatory T (Treg) cells. These data demonstrate an advantage of TEC-derived GILT for antigen processing and subsequent T cell tolerance. Only chimeras that lacked GILT in both TECs and hematopoietic cells were protected from melanoma and had a high effector T cell:Treg cell ratio. Thus, GILT diminishes T cell-mediated protection from melanoma through promoting thymic deletion and Treg cells.
The use of alternative promoters in T cell development
Authors: Chiang Jeffrey , Ku Wai Lim , Cui Kairong , Zhao Keji , Hodes Richard J. 
Published: January 2023

Abstract
Many mammalian genes, including a number involved in immune development and function, use multiple promoters encoding expression of the same protein product. To understand the function of alternative promotors during T cell development, we carried out genome-wide RNA-Seq analysis of DN, DP, CD4 and CD8 thymocytes. 26% (2469/9595) of genes expressed by thymocytes used two or more alternative promotors at some point in T cell development. However, only 3.2% (80/2469) of the genes which utilized two or more promoters showed the difference of promotor activity among different T cell developmental stages which were validated by selected representative genes with qPCR assay. To analyze mechanisms of alternative promoter regulation, DNase hypersensitivity sites were mapped, and ChIP-Seq with anti-H3K4me3 antibodies was performed in the same thymic subpopulations. To determine the in vivo functional role of alternative promotors for T cell development, several genes were selected for study by deletion of individual promoters by CRISPR technology. We have initially generated mice with deletion of distal or proximal promotor of the lck gene and demonstrated distinct functional roles of the two promoters: the proximal lck promoter is critical for early stages of thymic T cell development while the distal promoter was necessary for normal T cell activation. Ongoing studies will characterize more broadly the functional role of alternative promoter expression in thymocyte development, and the mechanisms mediating selective promoter activity.
Control of T cell development and function by Protein Arginine Methyltransferase 5
Authors: Hagman James R. , Fleenor Courtney J. , Arends Tessa , Pandey Ahwan , Abraham Christopher G. , Dege Carissa L. , Straign Desiree , Danhorn Thomas , Reinhardt R. Lee , Espinosa Joaquin M. , O’Connor Brian P. 
Published: January 2023

Abstract
Temporally and spatially regulated gene expression is essential for the development of thymocytes and functional T cells. Gene transcription is controlled, in part, by reversible post-translational modifications (PTM) of histones by epigenetic regulators. In turn, histone PTMs are bound by reader proteins, which mediate transcriptional activation and repression. Together, PTMs and readers enhance or suppress the accessibility of DNA to sequence-specific DNA-binding proteins. Epigenetic mechanisms are often dysregulated in human diseases and are promising targets for cancer treatments and immunotherapy. One such candidate is Protein Arginine Methyltransferase 5 (PRMT5), a type II arginine methyltransferase. PRMT5 catalyzes symmetric dimethylation (Rme2s) of arginines on histones H2A, H3, and H4. Rme2s strongly correlates with transcriptional repression. However, the roles of PRMT5-dependent histone arginine methylation in thymocyte development and T cell activation are poorly understood. Dynamic expression of Prmt5 transcripts suggests that the methyltransferase modulates transcription at select stages in T lymphopoiesis. Previous studies employing hematopoietic system-wide depletion of Prmt5 detected reduced cellularity of T cell progenitor populations, but effects on thymocyte transcriptomes were not determined. Here, we describe consequences of the loss of PRMT5 on T cell progenitor stage-specific transcription. PRMT5 suppresses activities of key transcription factors by regulating DNA accessibility, which in turn prevents inappropriate transcription in thymocytes in vitro and in vivo. Together, our data support a role of PRMT5 as an essential ‘gatekeeper’ of T cell development and function.
Elevation of choline metabolism contributes to the prolonged B cell survival and expanded B cell compartment in the spleen of B cell-specific TRAF3-deficient mice
Authors: Gokhale Samantha , Zhu Sining , Lu Wenyun , Xie Ping 
Published: January 2023

Abstract
Specific deletion of the tumor suppressor gene TRAF3 from B lymphocytes (B-TRAF3−/−) in mice leads to prolonged survival of mature B cells and expanded B cell compartment in secondary lymphoid organs. To understand the metabolic basis of the prolonged survival of TRAF3−/− B cells, we performed LC-MS-based metabolome and lipidome screening with resting splenic B cells of young adult B-TRAF3−/− and littermate control (LMC) mice. Our metabolome screening data showed that phosphocholine (P-Cho) and phosphoethanolamine, two small metabolites and precursors of phospholipids, were significantly increased in TRAF3−/− B cells. Consistently, our lipidome screening revealed markedly elevated levels of multiple species of phosphatidylcholine (PC) and phosphatidylethanolamine in TRAF3−/− B cells. Interestingly, our transcriptome profiling identified Chkα, the rate-limiting enzyme of choline metabolism, as a significantly up-regulated gene in TRAF3−/− B cells. To investigate the functional importance of elevated choline metabolism, we examined the effects of two Chkα inhibitors, MN58B and RSM932A. We found that both MN58B and RSM932A potently induced cell apoptosis in TRAF3−/− mouse B lymphoma and human multiple myeloma cell lines in vitro. Furthermore, in vivo administration of MN58B and RSM932A partially decreased the spleen size and B cell compartment in B-TRAF3−/− mice. Together, our results indicate that TRAF3 specifically re-engineers the P-Cho-PC-PE metabolic pathways to regulate mature B cell survival. Our findings also suggest that the P-Cho-PC-PE metabolic pathways have diagnostic and therapeutic value for B cell malignancies with TRAF3 deletion or relevant mutations.
Cell Death Markers in Sepsis
Authors: Madiraju Charitha , Mallarpu Chandra Shekar , Prasad Sudhir , Singarapu Maneendra , Kim Jean H. , Haririparsa Neda , Bratic Nemanja , Brar Harvinder , Cashman John R. , Chelluri Lakshmi Kiran 
Published: January 2023

Abstract
Cell death mechanisms play an important role in the pathogenesis of sepsis. The goal of our study was to understand cell death regulatory mechanisms such as pyroptosis, necroptosis and apoptosis in sepsis. Our preliminary study design included patients diagnosed with sepsis that were treated in clinical settings. Patients diagnosed with sepsis (test) were compared with healthy individuals (control). Cell death-regulating markers were assessed in plasma, peripheral blood mononuclear cells (PBMCs) and PBMC cell lysate samples derived from patients (N =8) and healthy controls (N=8). Samples were analyzed by ELISA, flow cytometry and immunoblotting methods. Preliminary flow cytometry evaluation of PBMC samples from patients showed elevated levels of cell death markers when compared to samples from healthy individuals. Results from ELISA experiments showed both plasma and PBMC cell lysate samples from septic patients had an increase in a marker for necroptosis, in comparison with control samples (P &lt; 0.05 and P &lt; 0.1, respectively). Interestingly, test samples showed modulation in a marker for apoptosis and a marker for pyroptosis in comparison with controls (P &lt; 0.05). Preliminary results suggest that there exists distinct cell death mechanisms with respect to the clinical outcomes observed in septic patients.
Epigenetic Regulation of the COX-2/PGE2 Pathway in Macrophages Results in Impaired Wound Healing in Diabetes
Authors: Davis Frank M. , Joshi Amrita , Kimball Andrew , den Dekker Aaron D. , Schaller Matthew , Wilke Carol A. , Moore Bethany B. , Gallagher Katherine 
Published: January 2023

Abstract
Macrophage (Mϕ) plasticity, allowing for transition of Mϕs from an inflammatory to a reparative phenotype, is critical for normal wound healing. In pathologic conditions, such as type 2 diabetes (T2D), wounds fail to heal due to impaired resolution of inflammation. There is increasing evidence that epigenetic-based mechanisms, including DNA hypomethylation, control Mϕ function. The cyclooxygenase (COX)-2/Prostaglandin E2 (PGE2) axis as well as upstream pathways including cytosolic phospholipase (c(PL)A2), involved in the release arachidonic acid, have been associated with chronic inflammation, however they have not been examined in diabetic wounds. The purpose of this study was to investigate if epigenetic modifications alter gene expression of the COX-2/PGE2 and c(PL)A2 pathway leading to derangements in macrophage inflammation in diabetes. Using the murine diet induce obesity (DIO) model, we evaluated diabetic wound healing compared with control mice. We demonstrate that TGFβ is elevated in DIO wound Mϕs in comparison to control wound Mϕs. Further, the increase in TGFβ stimulates miR-29B production in wound Mϕs resulting in hypomethylation and overexpression of the COX-2 gene. This results in diabetic wound Mϕs displaying increased levels of COX-2 which shunts amino acid metabolism towards the COX-2 pathway increasing PGE2 while limiting leukotriene synthesis via 5-lipoxygenase (5-LO) pathways. Lastly, c(PL)A2 appears to be upregulated in diabetic wound Mϕs and human diabetic monocytes. In summary, these results demonstrate that the COX-2/PGE2 pathway is increased in diabetic wound Mϕs by elevated TGFβ/miR-29b and results in elevated PGE2 that drives Mϕ-mediated inflammation preventing wound repair.
Molecular mimicry and signaling by human monoclonal autoantibody derived from human myocarditis and heart failure may contribute to fibrosis and remodeling in cardiomyopathy
Authors: Myers Jennifer M. , Alvarez Kathy M. , Reim Sean , Bentley Rebecca , Garman Lori , Chen Allshine , Wiley Graham , Bebak Melissa , Montgomery Courtney G. , Gaffney Patrick , Simpson Kathleen E. , Stavrakis Stavros , Cooper Leslie T. , Cunningham Madeleine W. 
Published: January 2023

Abstract
Previous studies in animal models of myocarditis suggest that immune responses against cardiac myosin may lead to remodeling in the heart and dilated cardiomyopathy. In human myocarditis and dilated cardiomyopathy, anti-cardiac myosin autoantibodies significantly associated with myocarditis in cohorts of adults and children and reacted significantly with both human cardiac myosin (HCM) and the β-adrenergic receptors (βAR). We produced a human monoclonal antibody (mAb) derived from human myocarditis which reacted with HCM and specific peptides from pathogenic regions of HCM. Autoantibodies against HCM and βAR from myocarditis and dilated cardiomyopathy including the human myocarditis-derived mAb targeted heart cells by signaling protein kinase A (PKA) in a primary heart cell line. To further define potential pathogenic mechanisms of the human mAb and serum autoantibodies from myocarditis and cardiomyopathy and to test our hypothesis that autoantibodies may alter gene expression related to extracellular matrix, remodeling and fibrosis, we investigated genes differentially expressed by RNA sequence analysis of a primary heart cell line after treatment with the human mAb and sera from both children and adults with myocarditis. The overall heatmap of differentially expressed genes in heart cells demonstrated a significant difference between the control sera group and both human mAb and myocarditis sera treatment groups. We identified upregulated genes related to fibrosis, apoptosis, inflammation and cardiomyopathy, suggesting that the myocarditis- and cardiomyopathy-derived autoantibodies may induce signaling in primary heart cells and potentially contribute to fibrosis and remodeling in the heart.