bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of T cell function by butyrophilin 3A1 (CD277) (IRC8P.453)
Authors: Wang Jinghua , Manick Brian , Wu Guoping , Kalabokis Vassili 
Published: January 2023

Abstract
The butyrophilins have similar immunomodulatory functions to B7 family, and may represent a novel subset of co-regulatory molecules. However, the functions of these butyrophilin molecules remain to be identified. We expressed the human butyrophilin 3A1 (BTN3A1) extracellular domain human IgG1 Fc Chimeras in a mammalian cell line and investigated the biological functions of the purified recombinant BTN3A1 protein in vitro. In this study, human T cells were treated with plate-bound anti-CD3 and either BTN3A1/Fc, or control human IgG1 Fc. The levels of multiple cytokines were measured in cell culture supernates using R&D Systems Proteome Profiler™ Human Cytokine Array, and confirmed by measuring individual cytokines using cytokine-specific Quantikine® ELISA kits. BTN3A1 significantly reduced T-cell derived cytokines including IL-2 and IL-5. BTN3A1 also markedly inhibited anti-CD3-induced human T cell proliferation. Furthermore, BTN3A1 showed binding to anti-CD3 activated T cells, but not inactivated T cells. Moreover, recombinant human BTN3A1 modified TCR signaling to inhibit Foxp3 expression and regulatory T cell development in naïve CD4+ T cells. Taken together, our data suggest that BTN3A1 acts as a co-inhibitory molecule to modulate T cells through unknown receptor on the surface of activated T cells.
Histone methyltransferase, Setdb2, regulates wound healing in a diet-induced obesity model of diabetes (IRM9P.601)
Authors: Joshi Amrita , Allen Ronald , Kroetz Danielle , Schaller Matthew , Dalton Jennifer , Kunkel Steven , Gallagher Katherine 
Published: January 2023

Abstract
Impaired wound healing in Type 2 diabetes (T2D) is associated with a chronic inflammatory response mediated by accumulation of macrophages producing pro-inflammatory cytokines. While recent work has shown that epigenetic regulation plays a key role in influencing macrophage function, the specific alterations that occur in T2D macrophages and their effects on gene transcription have not been elucidated. Work in our laboratory has shown that Histone H3 Lys9 methylation that results in transcriptional silencing is mediated by Setdb2 (SET domain, bifurcated2) and plays a critical role in lung inflammation. Thus, we examined the role of Setdb2 in wound healing and macrophage activation in a murine model of T2D. Superficial wounding resulted in increased transcript levels of Setdb2 in wound macrophages of control mice compared with diabetic mice. Specific deletion of Setdb2 in macrophages (Setdb2lyz2) using cre recombinase resulted in delayed wound healing. Furthermore, bone marrow derived macrophages from Setdb2lyz2 cre+ mice show reduced expression of key regulatory mediators such as IFN-β. In summary, our data indicate that Setdb2 may play a protective role in wound healing by regulating inflammation.
Weight loss induced by blocking CB1 cannabinoid receptors is associated with an altered microRNA profile and suppression of inflammation in adipose tissue (IRM9P.458)
Authors: mehrpouya pegah , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Obesity is associated with chronic inflammation. Cells of the immune system express cannabinoid CB1 and CB2 receptors, the activation of which triggers anti-inflammatory effects. We investigated the effect of treatment of mice with SR141716A, a cannabinoid receptor1 (CB1) antagonist, on Diet-Induced Obesity (DIO), specifically whether such a treatment can induce anti-inflammatory state in adipose tissue. DIO model was generated by feeding C57BL/6J mice with high-fat diet (HFD) whereas their lean, age-matched controls were fed low-fat diet (LFD). HFD-fed mice were treated with either SR141716A (10mg/kg/day) or vehicle by daily oral gavage for 4 weeks starting at week 12. To assess the effect of SR141716A beyond its effect on weight loss and diet intake, pair-feeding was conducted in diet-intake matched controls and diet intake was adjusted in body-weight-matched controls. Our data demonstrated that SR141716A suppressed diet intake transiently however weight loss and reduction in fat mass was persistent. High throughput miR analysis showed that SR141716A skewed adipose tissue macrophage balance to more anti-inflammatory macrophages. Induction of myeloid-derived-suppressive cells and TH2 cells followed by suppression of pro-inflammatory macrophages were observed in SR141716A-treated HFD-fed group. Together, this study demonstrates that CB1 receptor blockade affects miR expression that induces anti-inflammatory state in adipose tissue independent of calorie intake inhibition.
An insight on antibody-mediated immune-metabolism in inflammation and homeostasis: the role of Fc gamma receptors in regulation of indolemaine 2,3 dioxygenase activity in dendritic cells (IRM9P.460)
Authors: Morsi Hadeil , Salazar Fabian , Ghaemmaghami Amir 
Published: January 2023

Abstract
Indoleamine2, 3-dioxygenase (IDO) is a key immune-modulatory enzyme in Tryptophan catabolism that is highly expressed by Dendritic cells (DCs). Notably, DCs are the main mediators of Fc-dependent anti-inflammatory effects of Intravenous Immunoglobulins (IVIgs) as well as Immune Complexes (ICs)-induced inflammation. Hence, we aimed to study the role of FcγRs in regulation of IDO activity in human DCs from healthy individuals, which remains elusive compared to the well-established role of TLRs. Plasma-pooled human IgG showed no effect on IDO activity in un-stimulated monocyte-derived DCs (Mo-DCs). However, on TLR4 stimulation, IgG down-regulates LPS-induced IDO activity. This counter-regulation was enhanced further, by blocking FcγRIIA, the highest expressed FcγR on Mo-DCs. Interestingly, this effect was elicited by both soluble and plate-bound IgG, resembling IVIgs and ICs, respectively, but with higher potency with soluble IgG. Strikingly, mouse IgG demonstrated a similar effect to human IgG. Herein, we propose a threshold model for IDO regulation through a crosstalk between TLR and FcγRs in homeostatic and inflammatory conditions. IgG was only able to down-regulates TLR-4-induced IDO activity in human DCs, with no effect in the steady state. Furthermore, data suggest that recognition of conserved proteins or glycan moieties of IgG by gamma associated receptors modulate IDO activity. Such insight has potential therapeutic implications for antibody-based immunotherapy.
Tripeptidyl peptidase II links intracellular amino acids homeostasis with immune response through glycolytic pathway (IRM9P.463)
Authors: Lu Wei , Zhang Yu , Hambleton Sophie , Lenardo Michael 
Published: January 2023

Abstract
Proteasomes and lysosomes constitute the major cellular proteolytic systems that not only remove unfolded, damaged, or unnecessary proteins, but also provide an essential supply of free amino acids for energy and new protein synthesis. Tripeptidyl peptidase II (TPP2) is a required component of the proteasome-ubiquitin protein degradation pathway. The patients lack of TPP2 manifest a combined immunodeficiency with recurrent infections, severe autoimmunity, and neurodevelopmental delay. We observed a marked increase in lysosome number and lysosomal proteolytic activity in patient cells lacking TPP2 activity. Upregulated lysosomes compensated for deficient amino acid homeostasis when TPP2 was absent. We confirmed these results in Tpp2 knockout mice and in normal human cells treated with butabindide, a selective chemical inhibitor of TPP2. However, we also found that the overabundant lysosomes exerted a pathogenic effect in which key glycolytic enzymes including hexokinase II (HKII), were consumed. The resulting decreased glycolytic potential in TPP2 defective cells selectively attenuated the production of several immunomodulatory cytokines such as IFN-γ and IL-1β, whose production require aerobic glycolysis. Taken together, our data reveal the vital role of TPP2 in amino acid supply and unveil the intimate homeostatic balance between lysosome number, amino acid level, and glycolysis in mammalian cells.
Expression pattern of hepcidin in E.coli challenged obese mice (IRM9P.600)
Authors: Lai DengMing , Chen QingHua , Zeng CongLi , Chen QiXing , Fang XiangMing , Shu Qiang 
Published: January 2023

Abstract
Background:Hepcidin is a master regulator of iron metabolism and plays important roles in the pathophysiology of sepsis. Obesity leads to host immune changes and is associated with the pathogenesis of sepsis. Increased hepcidin expression has been observed in adipose tissue. However, the expression pattern of hepcidin in obese mice after sepsis challenge remains unknown. Methods:Mice were fed with high-fat-diet (HFD) containing 60% energy from fat for 12 weeks. The control mice were supplied with a diet with 10% energy from fat. Sepsis was induced by intraperitoneal injection of E. coli. Hepcidin mRNA expression was studied in peripheral blood mononuclear cell (PBMC), liver, spleen, adipose tissue and peritoneal macrophage. Results:HFD-fed mice showed higher weight and increased total cholesterol and triglyceride levels in serum than controls. Obese mice, as compared with controls, exhibited increased mortality following E.coli administration. Bacterial load and IL-6 levels in blood and peritoneal lavage fluid in the HFD group were higher than controls. The hepcidin mRNA levels were increased both in HFD and LFD groups. Especially, in septic obese mice, the hepcidin mRNA levels in PBMC, spleen and peritoneal macrophage, but no in liver and adipose tissue, were higher than those in controls. Conclusions:The different expression pattern of hepcidin after E.coli challenge suggests that tissue specific hepcidin may take a diverse role in the pathogenesis of sepsis in obese mice.
Significance of IL-15 in IL-4 induced B cell activation, immunoglobulin isotype class switching and IgE production (IRM10P.613)
Authors: Shukla Anshi , Rajavelu Priya , Upparahalli Venkateshaiah Sathisha , Jalahalli Mariswamy Siddesha 
Published: January 2023

Abstract
IgE-mediated immune responses contribute to the pathogenesis of allergic diseases. Despite their significant role in immune biology, the factors that produce and regulate IgE responses are yet to be established. Notably, IL-4 is a well-established cytokine involved in immunoglobulin (Ig) isotype class switching and IgE production. However we recently observed that IL-15 over expression in mice (pharmacologically or by transgene insertion) induces IgE levels. Therefore we hypothesized that IL-15 might have some role in B cells proliferation, activation, Ig isotype class switching and IgE production. Accordingly we examined the level of IgE, B cells and Ig isotype class switching in response to IL-15 using ELISA, real time qPCR and flowcytometric analysis. Our initial in vitro data show proliferation, activation and induced Ig class switching genes in purified B cells following rIL-15 treatments. Further, we show reduced IgE levels in IL-15 gene-deficient mice compared to Balbc mice following Aspergillus fumigatus exposure. Furthermore we examined the role of IL-15 in IL-4 mediated IgE production. We observed that IL-4 gene-deficient mice have increased IgE levels and B cell activation following intravenous administration of IL-15 compared to saline. However, significant IgE levels were observed following IL-15 treatment in Balbc mice compared to IL-4 gene-deficient mice.Together we conclude that IL-15 is required for the IL-4 induced B cell activation and allergen induced IgE.
The development og CD138high Breg in AAD is dependent on Semaphorin 4C (IRM10P.614)
Authors: Xue Di , Divangahi Maziar , Mazer Bruce , Fritz Jorg , Mindt Barbara , Beland Marianne , Fiter Ryan 
Published: January 2023

Abstract
We have previously found that mice with Semaphorin 4C (Sema4C) deficient B cells exhibited exaggerated airway damage and pulmonary inflammation during allergic airways disease (AAD). We aimed to determine the mechanisms by which Sema4C on B-cells regulates AAD. Following allergen exposure, Sema4C-/- AAD mice had significantly fewer CD138high IL-10+ B cells (CD138high Breg) and significantly increased CD138high IL4+ B cells in spleen. In vitro, Sema4C-/- B cells consistently failed to differentiate into CD138high Breg under Th2 culture conditions. Adoptive transfer of Sema4C-/- plasma cells induced exacerbated Th2 responses, with airway eosinophilia and increased IL-4+ inflammatory cell infiltrates in the lungs. In contrast, transfer of CD138high Breg to WT mice with AAD dramatically alleviated airway inflammation. Co-culturing Sema4C-/- B cells with Sema4C transfected cells (Hek293-Sema4C) corrected impaired CD138high Breg development. Co-culture of B cells with Hek293-Sema4C decreased IgE production for both WT and Sema4C-/- B cells. Sema4C is essential for the development of CD138high Breg and may play a crucial role in the auto-regulation of B cells in allergic inflammation.
Control of mammalian hematopoiesis and humoral immune response by microRNA-142 (IRM10P.617)
Authors: Boldin Mark , Kramer Nicholas , Wang Wei-Le , Reyes Estefany , Kumar Bijender , Chen Ching-Cheng , Chandran Ramakrishna , Cantin Edourad , Taganov Konstantin , Chau Nelson 
Published: January 2023

Abstract
MicroRNAs (miRNAs) are a class of powerful post-transcriptional regulators implicated in the control of diverse biological processes, including regulation of hematopoiesis and the immune response. To define the biological functions of miR-142, which is specifically and abundantly expressed in immune cells, we created a mouse line with targeted deletion of this gene. Our analysis of miR-142-/- mice revealed a critical role for this miRNA in the development and homeostasis of lymphocytes. Marginal zone B cells expand in the knockout spleen, while the number of T and B1 B cells in the periphery is reduced. Abnormal development of hematopoietic lineages in miR-142-/- animals is accompanied by a profound immunodeficiency, manifested by hypoimmunoglobulinemia and failure to mount a productive immune response to soluble antigens and virus. miR-142-/- B cells express elevated levels of BAFF receptor (BAFF-R) and as a result proliferate more robustly in response to BAFF stimulation. Lowering the BAFF-R gene dose in miR-142-/- mice rescued the B cell expansion defect, suggesting that BAFF-R is a bona fide miR-142 target through which it controls B cell homeostasis. Collectively, our results uncover miR-142 as an essential regulator of lymphocyte ontogenesis and suggest that lesions in this miRNA gene may lead to primary immunodeficiency.
FCRL5 regulates innate-like B cell natural antibody production and T cell-independent responses (IRM10P.607)
Authors: Becker Eugene , Zhu Zilu , Won Woong-Jai , Dalla-Favera Riccardo , Davis Randall 
Published: January 2023

Abstract
At homeostasis and during primary responses to pathogens, humoral host defense is chiefly coordinated by body cavity-derived B-1 and splenic marginal zone (MZ) B cells. These early responders have been termed “innate-like” due to their broadly-reactive natural antibody repertoires and rapid responses to T cell-independent (TI) antigens. However, what regulates these functions in these specialized cells remains unclear. Fc receptor-like (FCRL) molecules have homology to the classical FCRs for IgG and IgE, exhibit complex tyrosine-based regulation, and are preferentially expressed by B cells. Notably, these receptor-genes have been increasingly associated with immune disorders and human (h) FCRL3 has emerged as a risk factor in autoimmunity. To better understand hFCRL3’s role in B cells, we investigated its closest relative, mouse FCRL5 (mFCRL5), which shares homologous extracellular domains, binary tyrosine-based signaling, and innate-like B cell expression. Here we characterize the first in vivo model of FCRL function by examining Fcrl5 deficient mice. Our findings demonstrate that at homeostasis Fcrl5 deficient mice have normal total B cell numbers and subset frequencies, but enhanced natural antibody production. By contrast, Fcrl5 deficiency was found to impair TI humoral responses, but T cell-dependent responses remained intact. These data indicate a critical role for mFCRL5 in regulating innate-like B cell function at the interface of host defense and tolerance.
The negative regulatory role of ORMDL3 in antigen-induced cytokine production, cyclooxygenase-2 expression, and chemotactic response in mast cells (IRM11P.637)
Authors: Draber Petr , bugajev Viktor , Halova Ivana , Stegurova Lucie , Draberova Helena , Bambouskova Monika , Paulenda Tomas , Draberova Lubica 
Published: January 2023

Abstract
Single-nucleotide polymorphism studies have linked the chromosome 17q12-q21 region, where human ORMDL3 gene is localized, to the risk of asthma and other inflammatory diseases. Although mast cells are involved in development of these diseases, the contribution of ORMDL3 to mast cell physiology is unknown. In this study we examined the role of ORMDL3 in mouse bone marrow-derived mast cells with reduced or enhanced ORMDL3 expression. Our data show that in antigen-activated mast cells, reduced expression of the ORMDL3 protein had no effect on degranulation, calcium response, and sarco/endoplasmic reticulum Ca2+ ATPase 2b expression, but significantly enhanced phosphorylation of the IκBα and translocation of the NF-κB p65 subunit into the nucleus. These events were followed by an increased expression of pro-inflammatory cytokines (TNF-α, IL-6, and IL-13), chemokines (CCL3 and CCL4), and cyclooxygenase-2, an enzyme involved in the synthesis of prostanoids. Antigen-mediated chemotaxis was also enhanced in ORMDL3-deficient cells, whereas adhesion and spreading on fibronectin were decreased. On the other hand, increased expression of ORMDL3 had no significant effect on mast cell signaling events, except for reduced antigen-mediated chemotaxis. Our data provide the evidence that ORMDL3 in mast cells acts as a non-inducible suppressor of NF-kB-directed signaling pathways and chemotaxis.
Lipoteichoic acid of Lactobacillus plantarum inhibits Staphylococcus aureus-induced IL-8 expression in human intestinal epithelial cells (IRM11P.641)
Authors: kang Seok Seong , Noh Su Young , Yun Cheol-Heui , Han Seung Hyun 
Published: January 2023

Abstract
Lactobacilli as probiotics are well known to alleviate intestinal inflammation. However, the molecular basis for the mechanism has not been well understood. Here, we investigated the inhibitory effect of Lactobacillus plantarum lipoteichoic acid (Lp.LTA) on the induction of interleukin-8 (IL-8), which plays an important role in inflammatory responses by recruiting various immune cells, in human intestinal epithelial cells, Caco-2. Staphylococcus aureus but not the lipoprotein-deficient S. aureus mutant, induced IL-8 expression, implying that the lipoproteins are crucial in the induction of IL-8 by S. aureus. Pam2CSK4, a synthetic lipopeptide that is known as a mimic of Gram-positive bacterial lipoproteins, also induced IL-8 expression in Caco-2 cells. While heat-inactivated L. plantarum or peptidoglycan of L. plantarum did not inhibit Pam2CSK4-induced IL-8 expression, Lp.LTA significantly inhibited its expression. Deacylated or dealanylated Lp.LTA failed to inhibit Pam2CSK4-induced IL-8 expression, suggesting that both lipid and D-alanine moieties are responsible for the inhibitory potency of Lp.LTA. Furthermore, Lp.LTA inhibited the activation of Toll-like receptor 2 sensing bacterial lipoproteins and the intracellular signal transduction involving p38 kinase, JNK and NF-κB in Pam2CSK4-stimulated Caco-2 cells. Taken together, these results suggest that Lp.LTA alleviates S. aureus-induced intestinal inflammatory responses by inhibiting IL-8 expression.
TNFα-mediated I kappa kinase phosphorylation of ITCH impairs ubiquitin ligase activity (IRM11P.625)
Authors: Perez Jessica , Abbott Derek 
Published: January 2023

Abstract
The I Kappa Kinase (IKK) signalosome serves as a central hub for the relay of inflammatory signaling. Once activated, the kinase components of this hub, IKKα and IKKβ, can phosphorylate substrates outside of the known NF-κB inflammatory cascade. Additionally, two other closely related kinases, IKKε and TBK1, share kinase homology with IKKα and IKKβ, but phosphorylate substrates related to IFN regulatory factor (IRF) activation. Despite their well-known roles in NF-κB and IRF activation, we and others have found substrates outside of these signaling cascades. Given this, we have devised a proteomic and bioinformatic approach to identify novel IKK substrates. In the present work, we show IKK-mediated phosphorylation of ITCH, an E3 ubiquitin ligase implicated in the maintenance of mucosal immunologic homeostasis. We identify the site of phosphorylation, and demonstrate the inactivating effect of IKK phosphorylation on ubiquitin ligase activity. Given that mice null for ITCH develop pulmonary interstitial fibrosis and gastritis and given that TNF strongly activates the IKKs and this phosphorylation event, we bred ITCH-/- mice with TNFR1-/- mice to determine if loss of TNF activation of IKKα/β could reverse this phenotype. Both mortality and lung inflammation were improved in TNFR1-/-ITCH-/- when compared with ITCH-/- mice. These results show that bioinformatic and proteomic studies can be coupled with mucosal immunology for novel insights.
Measurement of IL-6 levels in live cells using RNA detection probe with Imaging Flow Cytometer
Authors: Vaidyanathan Shobana , Friend Sherree , Weldon Don , Morrissey Philip 
Published: January 2023

Abstract
Acute inflammatory responses to infection and tissue damage are orchestrated by release of cytokines, primarily from macrophages and mast cells, but also by endothelium and tumor cells. A key initiator of the inflammatory response is tumor necrosis factor alpha (TNF-alpha), which binds to its receptor on target cells and activates a pro-inflammatory transcriptional program via the transcription factor NFkB. A critical downstream target of NFkB is the gene encoding interleukin-6, which stimulates an array of anti-infection processes, including synthesis of acute-phase proteins by the liver, proliferation of B cells, neutrophil production in bone marrow, and differentiation of Th17 helper T cells. Although beneficial in the context of attacking infection, dysregulation of IL-6 plays a deleterious role in autoimmune disease and cancer. In this study, we introduce a new method for measuring NFkB translocation by looking at the increase in IL-6 mRNA levels in the THP-1 suspension cell line using SmartFlare™ RNA detection probes. The ability to detect RNA in live cells and to acquire multi-spectral images of large numbers of cells allows for the accurate assessment of IL-6 mRNA levels in THP-1 cells.
In vivo consequences of inducible cell death via direct activation of RIPK3
Authors: Olsen Tayla M , Orozco Susana , Oberst Andrew 
Published: January 2023

Abstract
Necroptosis is a form of programmed cell death and is distinct from apoptosis in its mechanism and morphology; apoptosis is thought to be a non-inflammatory form of cell death whereas necroptosis is thought to be inflammatory. Necroptosis occurs in response to viral infection and is important for control of DNA and RNA viruses in vivo. While much of the necroptotic pathway has been characterized in vitro, the effects of necroptosis are poorly understood in vivo.
We recently described a system by which necroptosis can be triggered using a version of the pro-necroptotic kinase RIPK3 fused to a ligand binding domain. Here, we characterize a novel transgenic model in which this inducible form of RIPK3 is expressed in vivo, allowing induction of necroptosis in primary tissues. We find that widespread activation of necroptosis in vivo using this model triggers sepsis-like systemic inflammation.