bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of α-enolase on the monocyte of rheumatoid arthritis via induction of inflammatory cytokines (112.14)
Authors: Bae Seyeon , Kim Hye Min , Kong Joo Myoung , Yu Yeon Sil , Kim Hang-Rae , Hwang Young Il , Kang Jae Seung , Lee Wang Jae 
Published: January 2023

Abstract
α-enolase (ENO1) is expressed on the surface of activated peripheral blood mononuclear cells. Although it plays an important role on the development of B cells in germinal center, there are some findings regarding the roles of ENO1 on the pathogenesis of cancer. Recently, we found that the production of inflammatory mediators from concanavalin A-activated PBMCs was increased by ligation of ENO1with anti-ENO1 mAb. Therefore, we investigate the roles of ENO1 on the regulation of inflammation and chronic inflammatory disease, rheumatoid arthritis (RA). First, we examined the expression of ENO1 on the surface of PBMCs, synovial fluid mononuclear cells (SFMCs) and synovial tissue (ST) derived from RA patients. The surface expression of ENO1 was higher on PBMCs and SFMCs derived from RA patients than those from osteoarthritis (OA) patients or normal samples. And it was mainly expressed on CD14-positive monocyte/macrophages. Interestingly, the expression of ENO1 on cells from RA patients is dependent of the severity of disease. The production of inflammatory mediators was dramatically increased by the stimulation of ENO1 via the activation of p38MAPK and NF-κB. Taken together, our results strongly suggest that ENO1 is a critical factor for the induction of severe inflammation in RA via the induction of disease-related pro-inflammatory cytokine production.
Inhibition of vascular endothelial growth factor receptor reverses type 1 diabetes in nonobese diabetic mice. (112.17)
Authors: Villalta Armando , Lang Jiena , Kabre Beniwende , Kubeck Sam , Arch Robert , Bluestone Jeff 
Published: January 2023

Abstract
Small molecule receptor tyrosine kinase inhibitors (RTKI) represent a novel therapeutic avenue for the treatment of type 1 diabetes (T1D). These potential therapies target multiple kinases raising questions as to the precise target for intervention. In this investigation, we used a pharmacological approach to test whether vascular endothelial growth factor receptors (VEGF-R) participate in the regulation of disease pathogenesis in T1D in nonobese diabetic (NOD) mice. Treatment of new-onset diabetic NOD mice with PF-337210, an RTKI that inhibits the VEGF-R and platelet-derived growth factor receptors (PDGF-R), reversed diabetes. However, PDGF-R selective inhibitors did not, suggesting that VEGF-R are the principal targets mediating diabetes reversal. RTKI treatment of diabetic mice reduced insulitis, preserved islet mass and improved glucose tolerance, but had no effect on insulin resistance. Immunohistochemical studies suggested that the RTKI targeted VEGF-R expressed by the islet vasculature and not by leukocytes, suggesting that these drugs have a predominant effect on inflammatory processes that contribute to the pathogenesis of T1D rather than a direct effect on diabetogenic leukocytes. Collectively, the results show the inhibition of VEGF-R2 signaling can reverse diabetes by dampening inflammatory events that lead to islet dysfunction, and support the use of VEGF-R specific antagonists as novel therapeutics for the treatment of T1D.
Activation of endothelial roundabout receptor 4 (Robo4) reduces the severity of virus induced keratitis (112.2)
Authors: Sharma Shalini , Mulik Sachin , Rouse Barry 
Published: January 2023

Abstract
Anti angiogenic molecules exert a feedback control to restrain pathological angiogenesis, which include physical binding or inhibition of angiogenic signaling in blood vessel endothelial cells. The latter is the case when Slit2 ligand dependent activation of the blood vessel endothelial cell receptor roundabout 4 (Robo4) occurs. In this study, we demonstrate that Robo4 receptors are up regulated following herpes simplex virus (HSV) infection of the eye on the majority of the new blood vessel endothelial cells that occur in the corneal stroma. However, expression levels of the ligand for Robo4 receptors, Slit2, was not significantly increased during the disease process and the knockdown of Slit2 gene expression using lentiviral shRNAs had no effect on the extent of pathological angiogenesis. In contrast, providing additional Slit2 protein by subconjunctival administration resulted in significantly reduced angiogenesis. The Slit2 binding to Robo4 was shown to block the downstream VEGF signaling molecules Arf 6 and Rac 1 and reduced the anti-apoptotic molecule Bcl-xL in blood vessel endothelial cells. Our results indicate that augmenting the host Robo4/Slit2 system could provide a useful therapeutic approach to control pathological angiogenesis associated with HSV induced stromal keratitis (SK).
Interaction of suppressor of cytokine signaling 2 with phosphorylated Pyk2 regulates human NK cell function (113.1)
Authors: Jung Haiyoung , Lee Suk Hyung , Park Young-Jun , Yoon Suk Ran , Choi Inpyo 
Published: January 2023

Abstract
Natural killer cells are large granular lymphocytes that play an important role in innate immunity, contributing to the elimination of tumor or virus-infected cells. Resting NK cells, however, have only minimal cytolytic activity and secrete a low level of IFN-γ. The cytokine IL-15 can promote the expression of effectors in NK cells. In this study, we demonstrate that suppressor of cytokine signaling 2(SOCS2) has a novel role in IL-15-primed human NK cell function. SOCS2 expression was upregulated in NK cells following stimulation with IL-15. During IL-15 mediated NK cell priming, SOCS2 interacted with phosphorylated proline-rich tyrosine kinase 2(Pyk2) at tyrosine 402(p-Pyk2Tyr402) and induced the proteasome-mediated degradation of p-Pyk2Tyr402 via ubiquitination. Knockdown of SOCS2 resulted in the accumulation of p-Pyk2Tyr402 and blocked NK cell effector functions. In addition, NK cell cytolytic activity and IFN-γ production were inhibited by over-expression of the wild-type of Pyk2 but not by the over-expression of tyrosine 402 mutant of Pyk2. These results suggest that SOCS2 regulates human NK cell effector functions via control of phosphorylated Pyk2 depending on IL-15 existence.
CD163/WC1 functions as a pattern recognition receptor and co-receptor for the γδ TCR (113.18)
Authors: Hsu Haoting , Cheng Chuang , Baldwin Cynthia , Telfer Janice 
Published: January 2023

Abstract
CD163/WC1 proteins, specifically expressed by γδ T cells, are members of the scavenger receptor cysteine-rich (SRCR) domain family. Phosphorylation of a tyrosine in the WC1 cytoplasmic domain upon co-ligation of WC1 and TCR is required for the WC1 co-receptor activity. Two out of fourteen WC1 proteins contribute to the γδ T cell response to the spirochete Leptospira. Other closely related SRCR family members (e.g. DMBT1, CD6 and CD163A) bind to bacteria, suggesting that WC1 acts a signaling co-receptor in γδ T cell activation via Leptospira ligation. Jurkat cells expressing the chimeric protein CD4/WC1cyt, CD4/WC1cyt mutated on a dileucine endocytic motif, and CD4/WC1cyt mutated on two serines upstream of the dileucine motif were treated with PMA, resulting in endocytosis of wild type protein. The leucine mutants reduced PMA-induced endocytosis and enhanced IL-2 production stimulated by co-crosslinking of CD3 and CD4/WC1cyt, suggesting that sustained membrane co-ligation with CD3 increases WC1 signaling. The serine mutation had no effect on endocytosis, but did decrease IL-2 production, suggesting that serine phosphorylation of the cytoplasmic domain of WC1 is involved in T cell signaling. The N-terminal SRCR domain of the WC1 protein previously implicated in the Leptospira response binds strongly and specifically to Leptospira, supporting a model in which WC1 acts both as a pathogen recognition receptor (PRR) and a membrane proximal signaling co-receptor to the γδ TCR.
Identification of a novel immunosuppressive dendritic cell population derived from conventional DC (113.5)
Authors: Mikhailova Anastassia , Diao Jun , Cattral Mark 
Published: January 2023

Abstract
Dendritic cells (DC) play a central role in the initiation and regulation of immune responses. Conventional DC (cDC) arise from circulating immediate precursors (pre-cDC) and are currently thought to be terminally differentiated. Here we show, however, that cDC are capable of generating progeny that lost all characteristic features of cDC; and investigate the functional properties of this novel population. Sorted bone marrow pre-cDCs were cultured on a stromal monolayer in the presence/absence of granulocyte-macrophage colony stimulating factor (GM-CSF). On day three, proliferating CD11c+ MHCII+ cDC developed in both cultures. However, in the absence of GM-CSF, these cells gave rise to a homogeneous population of CD11clow MHClow cells (DC-regs) on day 8-10 of culture. In contrast to cDCs, DC-regs failed to up-regulate MHCII and co-stimulatory molecules in response to DC maturation stimuli. DC-regs displayed higher antigen capture capacity but were poor stimulators in T cell proliferation assays. Further, DC-regs not only induced T cell anergy, but also suppressed T cell proliferation in cultures containing immuno-stimulatory DC. Co-transfer of DC-regs with DCs in vivo inhibited T cell proliferation. DC-regs expressed know immunosuppressive molecules such as iNOS, arginase, ICOSL and PD-L1. Our findings challenge the prevalent view that cDC are terminally differentiated, and reveal their potential to generate a regulatory DC population with potent immunosuppressive properties.
Regulation of skin inflammation and T cell activity by annexin A1 (114.3)
Authors: Yang Yuan , Deane James , Song Wuqi , Hickey Michael , Morand Eric 
Published: January 2023

Abstract
Annexin A1 (Anxa1) is a mediator of the anti-inflammatory actions of glucocorticoids. We explored the role of Anxa1 in regulation of the T cell-mediated contact sensitivity (CS) model of skin inflammation. CS was induced in Anxa1-/- and wild type mice by sensitisation to oxazolone at day 0 and subsequent challenge with oxazolone at day 5 (first challenge) and day 7 (second challenge). Deficiency of Anxa1 significantly increased CD4+ T cell proliferation in the draining LN after the first challenge, although there was no significant difference in CS-induced skin edema between WT and Anxa1-/- mice. In contrast, a significant increase in skin edema was observed in Anxa1-/- mice after a second CS challenge, accompanied by increased antigen-specific proliferation of CD4+ and CD8+ T cells obtained from spleen and skin. Anxa1-/- mice displayed significantly increased RORγT, and IL-17A mRNA in LN after a second challenge, indicating Anxa1 regulation of T cell activity in CS related to the Th17 driven immune response. Intravital microscopy studies showed that Anxa1 deficiency was associated with significantly increased CD4+ adhesion in dermal venules after the first challenge, and CD4+ and CD8+ adhesion after the second challenge. These data demonstrate that endogenous Anxa1 is a potent inhibitory regulator of T cell activity in skin inflammation, potentially via an inhibitory effect on development of the Th17 axis.
IL-2 controls the stability of Foxp3 expression in TGF-β-induced Foxp3+ T cells in vivo (115.11)
Authors: Chen Qian 
Published: January 2023

Abstract
Stimulation of naïve mouse CD4+Foxp3- T cells in the presence of TGF-β results in the induction of Foxp3 expression and T suppressor function. However, Foxp3 expression in these induced Treg (iTreg) is unstable raising the possibility that iTreg would not be useful for treatment of autoimmune diseases. To analyze the factors that control the stability of Foxp3 expression in iTreg, we generated OVA-specific iTreg from OT-II Foxp3-GFP knock in mice. Following transfer to normal C57BL/6 mice, OT-II GFP+ cells maintained high levels of Foxp3 for 8 days. However, they rapidly lost Foxp3 expression upon stimulation with OVA in IFA in vivo. This unstable phenotype was associated with a strong methylation of the Treg-specific demethylated region (TSDR) within the Foxp3 locus. Administration of IL-2/anti-IL-2 complexes expanded the numbers of transferred Foxp3+ iTreg in the absence of antigen challenge. Notably, when the iTreg were stimulated with antigen, treatment with IL-2/anti-IL-2 complexes stabilized Foxp3 expression and resulted in enhanced demethylation of the TSDR. Conversely, neutralization of IL-2 diminished Foxp3 expression resulting in decreased suppressor function of the iTreg in vivo. Our data suggest that stimulation with TGF-β in vitro is not sufficient for imprinting T cells with stable Foxp3 expression. Administration of IL-2 in vivo stabilizes Foxp3 expression and may prove to be a valuable adjunct for the use of iTreg for the treatment of autoimmune diseases.
Therapeutic effects of TGF-β-induced iTregs on established autoimmune diseases (115.3)
Authors: Lan Qin , Wang Julie , Brand David , Stohl William , Zou Hejian , Horwitz David , Liu Zhongmin , Zheng Song Guo 
Published: January 2023

Abstract
While recent studies reveled injection of nTregs has less therapeutic effects on established autoimmune diseases, it is less clear if iTregs can treat these diseases. We now reveal that unlike nTregs, injection of iTregs can markedly ameliorate established autoimmune arthritis and lupus. Both antigen-specific and polyclonally induced iTregs suppressed the established CIA. CIA mice given iTregs had significantly lower clinic scores than control groups. We found nTregs but not iTregs were converted into Th1/Th17 cells in vitro and in vivo in the inflammatory milieu, injection of iTregs to naïve and immune deficient mice displayed similar levels of Foxp3 stability as comparing with nTregs. Of note, the stability of Foxp3 expression was only found in iTregs during established CIA. iTregs suppressed Th17 and osteoclast differentiation. Injection of iTregs to established lupus decreased levels of anti-dsDNA and proteinuria, and prolonged lupus survival. Anti-TGF-β Ab or TGF-βRI inhibitor, or anti-IL-10R Ab abolished the therapeutic effects of iTregs on lupus. We further observed that DC isolated from lupus received iTregs expressed lower levels of B7.1/7.2 and adoptive transfer of these DCs to another lupus mouse can suppress the disease development. We suggest that iTregs are stable and able to target DC in the inflammatory milieu, these DC then have become tolerogenic DC and further suppress disease progression via its direct or indirect effect in autoimmune disease settings.
Regulatory T cell perturbation in biliary atresia (115.4)
Authors: Tucker Rebecca , Brindley Stephen , Mack Cara 
Published: January 2023

Abstract
Biliary atresia (BA) is an inflammatory disease of the biliary system which leads to cirrhosis and the need for liver transplantation in the majority of children. The etiology of BA is unknown and one theory is that bile duct injury is due to a virus-induced, subsequent autoreactive T cell-mediated inflammatory insult. Perturbation of the quantity or function of regulatory T cells (Tregs) in BA could result in unchecked autoimmunity. The aim of this study is to determine if Tregs are diminished or dysfunctional in murine and human BA. Murine BA results from infection of BALB/c neonates with Rhesus rotavirus (RRV). Mice infected with RRV on day 1 that received either Tregs or control T cells on day 5 led to a difference in disease status as early as 4 days post-transfer. This trend continued and biliary disease occurred in 67% of controls vs. 39% of Tregs recipients at 17 days of life. Secondly, ablation of functional Tregs in 4 day-old mice by treatment with a CD25-depleting antibody followed by RRV infection 3 days later results in increased bile duct infiltrate and disease incidence at 3 weeks of age. Finally, analysis of human PBMCs revealed a decreased percentage of FoxP3+ Tregs in BA patients (1.5±0.3%) compared to age-matched controls (4.1±0.7%). These studies suggest that lack of or dysregulation of Tregs is present in the murine model of BA and that diminished numbers of Tregs may be implicated in human BA progression.
Asp299Gly polymorphism decreases TLR4-elicited activation of MyD88- and TRIF-dependent signaling pathways (116.17)
Authors: Figueroa Leandra , Medvedev Andrei 
Published: January 2023

Abstract
Innate immune cells detect Gram-negative bacterial lipopolysaccharide (LPS) via Toll-like receptor (TLR) 4 that employs myeloid differentiation primary response gene (MyD) 88 and TIR-domain containing adapter-inducing IFN-β (TRIF) pathways to activate NF-κB, IRF3, and induce pro-inflammatory cytokines and type I IFNs, respectively. Asp299Gly and Thr399Ile TLR4 polymorphisms have been associated with an increased risk of Gram-negative sepsis and certain bacterial infections. The mechanisms by which these mutations impact TLR4 signal transduction are incompletely understood. We used transfection-based complementation of TLR4-/- HEK293 cells with CD14, MD2, wild-type (WT) or polymorphic YFP-TLR4s to determine the impact of the Asp299Gly and Thr399Ile polymorphisms on LPS binding and TLR4-elicited activation of MyD88 and TRIF pathways. In contrast to strong LPS responses induced by WT TLR4, the Asp299Gly variant elicited decreased phosphorylation of p38 and IRF3, activation of NF-κB- and IFN-β-driven reporters, and expression of IL-8 mRNA. 293/MD2 cells expressing WT or polymorphic TLR4s showed comparable LPS binding, and had similar total levels of transfected YFP-TLR4s and Flag-MD2, as well as similar expression of total p38 and β-tubulin. Thus, the D299G polymorphism compromises TLR4-mediated activation of MyD88 and TRIF-dependent pathways, and these signaling deficiencies are not due to lower expression levels of the TLR4 receptor complex.
Sensing bacterial viability for host defense (116.23)
Authors: Sander Leif , Davis Michael , Boekschoten Mark , Dascher Christopher , Ryffel Bernard , Amsen Derk , Swanson Joel , Mueller Michael , Blander Julie 
Published: January 2023

Abstract
The innate immune system safeguards the host from invading microorganisms by eliciting carefully measured responses. For example, weaker responses are mounted to dead than to viable microorganisms, a property with important implications for vaccination and host defense. Pathogen associated molecular patterns (PAMPs), which serve to alert the immune system, are present in dead and viable bacteria, raising the question as to how the immune system discriminates between the two. Here we show that the innate immune system can directly sense bacterial viability through detection of a special class of viability-associated PAMPs (vita-PAMPs). We identify prokaryotic messenger RNA (mRNA) as a vita-PAMP present only in viable bacteria, recognition of which elicits a unique innate immune response. Detection of vita-PAMPs in phagocytosed bacteria induces robust production of IFN-β and activation of the NLRP3 inflammasome in various innate immune cells. All responses require the Toll-like receptor adaptor protein TRIF. Importantly, recognition of vita-PAMPs in vivo induces host protective antibody mediated immunity. Thus, the immune system actively gauges the infectious risk by scouring PAMPs for signatures of microbial life and thus infectivity. Detection of vita-PAMPs triggers an alert mode not warranted for dead bacteria. Vaccine formulations that incorporate vita-PAMPs could combine the superior protection of live vaccines with the safety of dead vaccines.
Ribozyme mediated alternative poly-adenylation of the inhibitory NK-ligand Clec2d. (116.36)
Authors: Horan Lucas 
Published: January 2023

Abstract
Structured RNAs that are embedded in mature mRNAs have been shown to effectively regulate gene expression in a variety of single-celled organisms. Our discovery of conserved mammalian hammerhead ribozymes located in the 3‘-untranslated regions (UTR) of certain C-type lectin type II (CLEC2) genes suggests that a similar form of regulation may also be operative in more complex organisms. The ribozyme-associated Clec2d gene codes for a broadly expressed cell-surface ligand, CLEC2D, that is involved in suppressing natural killer cell-mediated lysis of CLEC2D expressing cells. Published data show that CLEC2D is downregulated in infected cells and cells exposed to various forms of stress. We hypothesize that the location of the fast cleaving ribozyme in the 3‘UTR of a dynamically expressed gene implicates the ribozyme in regulating CLEC2D expression. Previously described regulatory ribozymes (both naturally occurring and engineered) induce loss of gene expression through ribozyme cleavage and stabilize gene expression by preventing ribozyme cleavage. Surprisingly we do find that cells expressing CLEC2D protein predominantly contain transcripts that are poly-adenylated at the ribozyme cleavage site. This directly implicates the mClec2d ribozyme in an alternative poly-adenylation mechanism and suggests ribozyme cleavage can be involved in stabilizing protein expression.
Evolving function of Toll-like receptor pathways (116.6)
Authors: Charette Jeremy , Phennicie Ryan , Pietraszewski Matthew , Lage Christopher , Smalley David , Kim Carol 
Published: January 2023

Abstract
The innate and adaptive arms of immunity work synergistically to mount robust host defenses against invading pathogens. The zebrafish lacks a fully functional adaptive immune system for the first four to six weeks of development, making it an ideal model for studying many aspects of innate immunity, including the Toll-like receptor (TLR) pathways. In mammals, TLRs recognize specific pathogen associated molecular patterns and, acting through adaptor molecules such as MYD88 and TICAM1, elicit an array of pro- and anti-inflammatory responses. In two recent studies we described differences in TLR ligand recognition and downstream signaling in the zebrafish, and proved that sequence homology does not necessarily indicate functional homology. These data support the notion that alternative signaling pathways may have evolved in the zebrafish. To characterize this system, ligands of the zebrafish TLRs and the intracellular protein-protein interactions were studied. Here we report the identification of unique interactors and targeted morpholino knock-down of TLR-associated genes to analyze the impact on signaling of each each protein-protein interaction within zebrafish TLR pathways. The discovery of novel protein interactions in the zebrafish highlights the potential of this model system for studies of innate immune function and further elucidates the evolution of vertebrate immunity.