bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structural basis of DAP12-associated intramembrane immunoreceptor assembly. (138.16)
Authors: Call Matthew , Wucherpfennig Kai , Chou James 
Published: January 2023

Abstract
Many of the molecular sensors that alert lymphocytes to dangerous events such as infection or transformation are modular receptors constructed from ligand-binding subunits and signaling subunits that assemble through specific contacts within the lipid bilayer to form functional complexes. Structure-function studies focusing on the extracellular and cytoplasmic domains of activating immunoreceptors like the T cell receptor (TCR)-CD3 complex have yielded a vast store of knowledge on the mechanisms governing ligand binding and the resulting intracellular biochemical cascades. However, the mechanisms physically linking these events across the membrane to initiate immune activation remain poorly defined, and structural information on the membrane-embedded portions of receptor complexes will be critical for developing more precise models. We previously published the first structure of an immunoreceptor-associated transmembrane (TM) signaling module, the TCR-associated ζζTM dimer. We now present the NMR structure of DAP12, a functional homologue of ζζ that provides the ITAM-based signaling capabilities to more than a dozen activating receptors expressed by NK cells and other hematopoietic lineages. We have also solved a structure of the trimolecular complex DAP12-TM forms with the NK cell receptor NKG2C. These structures reveal features that allow DAP12 to assemble with a multitude of different receptors with disparate TM sequences through focused polar contacts within the membrane.
Modulation of cytokine and nitric oxide production by keratinocytes and mononuclear phagocytes in a model of inflammatory acne. (138.26)
Authors: Watson Michelle , Ping E Ya , Lee Wei Li , Nowakowski Maja 
Published: January 2023

Abstract
Cytokines (TNFα, IL-1α, IL-6, IL-8, and IL-10) mediate the inflammatory process in various skin conditions, including acne. An in vitro co-culture model system involving different cell types was used to study the effect of UVB and other treatments on the inflammatory process, with nitric oxide (NO) and tumor necrosis factor (TNFα) as markers of inflammation. Cultured macrophages were stimulated with bacterial lipopolysaccharide (LPS) and exposed to UVB radiation (50 - 200mJ/cm2). The expression of nitric oxide (NO) was measured by modified Griess assay and the expression of TNFα was measured by quantitative ELISA. For co-culture systems, SC monocytes were combined with irradiated human epidermal keratinocytes (NHEK) and cultured macrophages were combined with peripheral blood mononuclear cells (PBMC). Samples of culture supernatants were collected at 1-6 days. When NHEK were irradiated with UVB (290-320 nm), a dose-dependent stimulation of TNFα production was observed (33-200% increase), but NO production was unchanged. In contrast, when macrophages were irradiated with UVB, significant inhibition of NO production (40% suppression, p&lt;0.001) was seen. The macrophage-PBMC co-cultures showed no alteration in TNFα production, whereas NO production showed a dose- and time-dependent increase. These data indicate that keratinocytes and mononuclear phagocytes have distinct products and responses to UVB radiation. This co-culture system is an improved model of cutaneous inflammation.
Analysis of cross-species IgG binding to human and mouse Fcgamma receptors (FcγRs) (138.29)
Authors: Shashidharamurthy Rangaiah , Bozeman Erica , Patel Jaina , Kaur Ramneet , Meganathan Jeyandra , Selvaraj Periasamy 
Published: January 2023

Abstract
Monoclonal antibodies from various species are in therapeutic and diagnostic use and are used extensively in cell depletion studies in rodents. Therefore, in the present investigation the cross-species IgG binding to FcγRs were performed using recombinant dimeric soluble form of human and mouse FcγRs. We observed that human CD16A was able to bind to rabbit IgG better than human CD32AH and CD32AR. The binding specificity is hCD16&gt;hCD32AH&gt;&gt;hCD32R. Both hCD32A alleles were able to bind mouse and rat IgG subtypes, whereas hCD16 binds weakly. hCD32AR binds mouse IgG1, IgG2a and IgG2b but not IgG3 (IgG1&gt;&gt;IgG2a≥IgG2b) whereas CD32AH binds IgG2a and IgG2b (IgG2a≥IgG2b) only not IgG1 and IgG3. With rat IgG subtypes, hCD32AR binds IgG1&gt;IgG2a&gt;&gt;IgG2b and hCD32AH binds IgG1&gt;IgG2b&gt;IgG2a whereas neither hCD32A alleles bound to IgG2c. We observed that hCD32AR allele binds stronger than hCD32AH to rat IgG. With human IgG subtypes, both hCD16A and hCD32AR binds IgG3&gt;IgG1&gt;&gt;&gt;IgG2≥IgG4, whereas hCD32AH binds IgG3&gt;IgG1&gt;IgG2&gt;&gt;&gt;IgG4. We have also characterized binding specificity of mouse FcγRs (mCD16A, mCD32B and mFcRIV) to rabbit and mouse IgGs. All three mFcγRs binds to rabbit IgG. For mouse IgG subtypes, mCD16A binds IgG2b&gt;IgG2a≥IgG1, mCD32B binds IgG1≥IgG2b&gt;IgG2a, whereas FcRIV binds IgG2a≥IgG2b only, none of the mFcγR bound to mIgG3. In conclusion, cross-species FcγR binding to different IgGs must be taken into consideration when IgG based therapeutics and diagnostics are evaluated in rodents.
HIGH DOSES OF TCR STIMULI PREVENT CONVERSION OF NAÏVE CD4+ T CELLS INTO iTREGS IN AN NF-kB-DEPENDENT MANNER (139.10)
Authors: Molinero Luciana , Alegre Maria-Luisa 
Published: January 2023

Abstract
Conversion of naïve CD4+ T cells into FoxP3-expressing regulatory T cells (iTregs) can be achieved in the presence of TGF-b and IL-2 upon TCR triggering by intermediate concentrations of antigen or anti-CD3 mAb, but is prevented by high concentration of antigen. The mechanism for this high dose inhibition of FoxP3 induction has not been elucidated. As TCR stimulation promotes NF-kB signaling, we hypothesized that TCR-induced NF-kB may be responsible for inhibition of FoxP3 expression at high TCR occupancy. Whereas increasing doses of anti-CD3 mAb or antigenic peptide resulted in inhibition of FoxP3 induction in wildtype T cells under iTreg conditions, CD4 transgenic cells expressing an NF-kB superrepressor retained the ability to express FoxP3. Conversely, overactivation of the NF-kB pathway via a constitutively active IKKb reduced Foxp3 expression in wildtype T cells at lower TCR occupancy. Furthermore, CARMA1, a key molecule for NF-kB signaling downstream of the TCR, was critical for Foxp3 inhibition at high TCR occupancy. This was not due to increased TGF-b- or IL-2-mediated signals, or diminished Akt activation in CARMA1-KO T cells, as phosphorylation of Smad2, STAT5 and the Akt target S6 was similar to that in wildtype cells. Rather, the accessibility of the Foxp3 locus was impaired in wildtype but not CARMA1-deficient T cells. Our results suggest that higher levels of NF-kB can inhibit FoxP3 chromatin remodeling, thereby preventing its induction.
Extracellular ATP Acts as a Danger Signal and May Contribute to Tissue Repair by Rapidly Stimulating P2RX7-dependent VEGF Release from Primary Human Monocytes (140.17)
Authors: Hill Lindsay , Gavala Monica , Lenertz Lisa , Bertics Paul 
Published: January 2023

Abstract
Extracellular ATP has been proposed to act as a danger signal to alert the immune system of cell damage. Release of high local concentrations of ATP activates the nucleotide receptor P2RX7, which triggers the release of pro-inflammatory mediators from monocytic cells. Although the pro-inflammatory actions of P2RX7 are well recognized, little is known regarding the potential function of P2RX7 in repair responses. Because the resolution of inflammation is characterized by monocytic cell-dependent production of pro-angiogenic factors, we evaluated the contribution of P2RX7 to this process. We observed that P2RX7 activation results in a time-dependent increase in VEGF mRNA; furthermore, both transient and long-term P2RX7 activation promote the robust release of VEGF from primary human monocytes. This VEGF release is calcium-dependent and associated with reactive oxygen species production. This previously unrecognized action of P2RX7 suggests that in addition to its established role in inflammation and cell death, P2RX7 signaling may also be important in the control of angiogenesis and wound repair.
Cannabidiol attenuates experimental autoimmune hepatitis by inducing accumulation of myeloid-derived suppressor cells (MDSCs) in liver (140.4)
Authors: Hegde Venkatesh , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Immune-mediated liver diseases are a major health problem worldwide. Cannabidiol (CBD), a non-psychoactive cannabinoid from hemp plant, modulates immune function and has shown therapeutic potential in treating inflammatory diseases. We studied the effects of CBD in a mouse model of autoimmune hepatitis induced by ConA. Administration of CBD after ConA challenge significantly suppressed hepatitis in a dose-dependent manner as shown by decreased aspartate transaminase levels and reduced liver injury. CBD also suppressed crucial inflammatory cytokines. Phenotypic analysis of infiltrating cells revealed that CBD acted by inducing arginase-expressing CD11b+Gr1+ myeloid-derived suppressor cells (MDSCs) in liver. Depletion of MDSCs using anti-Gr1 mAb reversed the protective effect of CBD. Furthermore, adoptive transfer of purified MDSCs in to naïve mice conferred partial resistance to ConA-hepatitis. CBD-induced MDSCs could effectively suppress T cell proliferation in vitro. Using vanilloid receptor-deficient Trpv1-/- mice, we observed that CBD acted independently of this receptor in the current model. Interestingly, CBD caused a robust induction of CD11b+Gr1+ MDSCs in the peritoneum which was completely blocked by PPAR-γ inhibitor and was attenuated in mast cell-deficient mice, suggesting that mechanistically, CBD may act through transcription factor PPAR-γ on tissue mast cells thus activating them to release specific mediators resulting in the migration and expansion of MDSCs.
A mouse model of food induced acute allergic angioedema (141.10)
Authors: Gangur Venu 
Published: January 2023

Abstract
Previous studies report that without the use of adjuvant such as cholera toxin, oral exposure to peanut proteins in mice results in oral tolerance. Here we tested the hypothesis that oral administration of peanut butter (instead of soluble peanut proteins) would be sufficient to sensitize C3H/Hej mice for peanut allergy. Groups of C3H/Hej mice were exposed to peanut butter or saline (orally, once per week) and examined for systemic IgE and IgG1 responses. Oral administration of peanut butter, but not saline, resulted in systemic peanut specific IgG1 and IgE antibody responses. Notably, as little as two oral exposures to peanut butter resulted in dramatic IgG1 antibody responses. Peanut butter elicited IgE and IgG1 antibodies were able to bind to peanut protein extract as well as to peanut butter preparation in ELISA system. Furthermore, oral challenge of sensitized mice, but not saline control mice, with peanut butter resulted in clinical symptoms of acute allergic angioedema with swollen lips, nostrils, face and eyelids. Mice also exhibited itching-scratching and rubbing symptoms with abnormal breathing. These data demonstrate for the first time that: oral peanut butter administration to C3H/Hej mice does not result in oral tolerance, but instead results in clinical sensitization for acute allergic angioedema. Funding: US EPA STAR Grant#R833133
TRANSCRIPTOME ANALYSIS OF MOUSE SPLEEN IN RESPONSE TO EGG OVOMUCOID TREATMENT (141.11)
Authors: Husain Mainul , Mine Yoshinori , Karrow Niel , Golovan Serguei 
Published: January 2023

Abstract
Food allergy is a serious health concern among infants and young children. Despite its growing prevalence, our current understanding of the molecular mechanism(s) of food allergy remains incomplete. In this study, transcriptome profile of spleen from BALB/c mice was analyzed using Affymetrix arrays. Female BALB/c mice 6-8 weeks of age were orally exposed twice weekly for 5 wks with 1 mg of egg ovomucoid (OVM) and 10 µg of cholera toxin (CT). Control mice were similarly sensitized with 100 µl of 10 µg/µl amino acid solution and CT. At wk 6, experimental and control mice were challenged by intraperitoneal (i.p.) injection with 1 mg of OVM and amino acid solution respectively. Mice were euthanized 40 min post challenge to collect the spleens. Total RNA extracted from spleen was subjected to microarray hybridization. Analysis of microarray data revealed 87 genes differentially expressed with at least ±1.5 fold (p≤0.05). Gene ontology (GO) analysis revealed several biological processes related to hypersensitivity, inflammation or immune response. Biological network analysis illustrated connections among some differentially expressed genes and several immune or hypersensitivity related processes. Expression of five differentially expressed genes from microarray experiment was validated by real-time RT-PCR. Results from this study revealed a network of genes that may enable us to better understand the underlying molecular mechanism(s) of food allergy.
A non-redundant role for Serpinb3a in the induction of mucus production in asthma (141.17)
Authors: Sivaprasad Umasundari , Askew David , Ericksen Mark , Gibson Aaron , Stier Matthew , Brandt Eric , Bass Stacey , Lindsey Mark , Daines Michael , Chakir Jamila , Stringer Keith , Wert Susan , Whitsett Jeffrey , LeCras Timothy , Aronow Bruce , Wills-Karp Marsha , Silverman Gary , Khurana Hershey Gurjit 
Published: January 2023

Abstract
Asthma is a major public health burden worldwide. Excessive mucus production and mucus plugging are key pathologic features of asthma, yet the mechanisms responsible for mucus production remain largely unknown and therapies to effectively target mucus hypersecretion are lacking. Using a murine asthma model, we showed that SerpinB3a, the mouse ortholog of the serine protease inhibitors, SERPINB4 and B3, contribute to house dust mite induced airway hyperresponsiveness (AHR), mucus production and goblet cell hyperplasia and expression of SPDEF, a transcription factor that mediates goblet cell differentiation. Microarray analysis revealed attenuated expression of multiple IL-13 regulated genes that contribute to mucus production in the Serpinb3a null mice and IL-13 treated mice showed attenuated AHR and mucus production. Our data have revealed a novel non-redundant role for SERPINB4 and B3 in mediating mucus production through regulation of SPDEF expression.
RGS16 regulation of Th2-mediated inflammatory responses (141.9)
Authors: Shankar Sucharita , Divietro Jeffrey , Xie Zhihui , Wilson Mark , Wynn Thomas , Druey Kirk 
Published: January 2023

Abstract
Chemokine receptors are G-protein coupled receptors (GPCRs) that direct immune/ inflammatory responses. Regulators of G-protein Signaling (RGS) proteins negatively regulate GPCRs by enhancing GTPase activity of Gα, catalyzing its deactivation. RGS16 in T lymphocytes regulates chemokine responses. Transgenic mice overexpressing RGS16 exhibited altered lymphocyte trafficking in an allergic pulmonary inflammation model compared to WT. Rgs16 mRNA increased upon naïve T cell activation with anti-CD3/anti-CD28. Lymph node (LN) T cells polarized towards Th2 in vitro expressed more Rgs16 than naïve cells. To further elucidate the role of RGS16 in Th2 cells, we generated Rgs16-/- (KO) mice. In chemotaxis assays, KO LN Th2 cells migrated more to CCL17 than WT cells, while cells from both groups migrated similarly to CXCL12 and CCL21. Mice were treated either mucosally or intravenously (IV) with Schistosome egg antigen (SEA), causing Th2-mediated eosinophilic pulmonary inflammation or granulomatous lung fibrosis, respectively. In the former model, bronchoalveolar lavage fluid from KO mice exhibited double the lymphocytes and eosinophils versus WT mice after tracheal challenge. In the IV model, granuloma size and liver inflammation markers increased in KO mice versus WT. In both models, KO LN cells secreted more Th2 cytokines (IL-4, 5, 13) versus WT after SEA re-stimulation. In summary, RGS16 in Th2 cells regulates lymphocyte activation and migration to Th2 chemokines.
Glabridin inhibits lipopolysaccharide-induced activation of a microglial cell line, BV-2, by blocking NF-κB and AP-1 (142.3)
Authors: Lee Ki Hoon , Park Sun Hong , Kang Jong Soon , Han Sang-Bae , Lee Kiho , Park Song-Kyu , Kim Hwan Mook 
Published: January 2023

Abstract
Glabridin, a flavonoid present in licorice root, is known to have anti-inflammatory and cardiovascular protective activities. In the present study, we report an inhibitory effect of glabridin on microglial activation. Glabridin dose-dependently attenuated lipopolysaccharide (LPS)-induced production of inflammatory mediators, including nitric oxide, tumor necrosis factor-α and interleukin-1β, in BV-2 cells, a murine microglia cell line. Moreover, mRNA expression of these inflammatory mediators was also suppressed by glabridin in LPS-stimulated BV-2 cells. Further study demonstrated that glabridin inhibited LPS-induced DNA binding activity of NF-κB and AP-1 in BV-2 cells. Collectively, the results presented in this report demonstrate that glabridin inhibits the production of inflammatory mediators in BV-2 cells and this is mediated, at least in part, by blocking NF-κB and AP-1 activation. Our results suggest that glabridin might be a potential therapeutic agent for the treatment of neuroinflammatory and neurodegenerative diseases.
Regulation of the expression of cyclooxygenases and production of PGI2 and PGE2 in human coronary artery endothelial cells by curcumin (142.5)
Authors: Tan Xiaoyu , Poulose Ellen , Zhu Bao-Ting , Stechschulte Daniel , Dileepan Kottarappat 
Published: January 2023

Abstract
Curcumin is a hydrophobic polyphenol present in turmeric (Curcuma longa). PGE2 and PGI2 are produced from arachidonic acid (AA) by the cyclooxygenases (COX1, COX2) and PGE2 synthase (PGE2S) and PGI2S. We evaluated the in vitro effect of CUR on COX1, COX2, PGI2S and PGE2S expression, and PGE2 and PGI2 production in human coronary artery endothelial cells (HCAEC). HCAEC were incubated with CUR (1.25-5.0 μM), and mRNA and protein expression were determined by qRT-PCR and Western blots, respectively. PGI2 and PGE2 levels in the culture supernatants were quantified by EIA. Incubation of HCAEC with CUR led to concentration-dependent increases in COX2 mRNA with a small but significant decrease in COX1 mRNA expression. CUR also induced 2- to 3-fold increase in PGI2S and PGE2S mRNA expression. Although CUR stimulated COX2, PGI2S and PGE2S mRNA, no increase in PGI2 or PGE2 production was noted. Addition of exogenous AA dose-dependently increased prostanoid production by both resting and CUR-treated cells. However, CUR-treated HCAEC produced significantly higher PGE2 when AA concentration was raised to 25µM. 14C-AA uptake by CUR-treated HCAEC was 30% lower than controls suggesting that CUR may inhibit exogenous AA uptake. These results show that CUR acts both as an inducer of COX2, PGI2S and PGE2S genes and a suppressor of COX1 gene. Taken together, these results suggest that CUR regulates AA metabolism and prostanoid homeostasis at both the gene and enzyme levels.
Early complement activation through alternative pathway significantly correlates to myocardial injury in patients undergoing cardiac surgery (142.6)
Authors: Zhang Ming , Hou Yunfang , Gulaya Karan , Lee Daniel , Lee Jeanie , Rushbrook Julie , Ko Wilson , Shevde Ketan 
Published: January 2023

Abstract
Global heart ischemia occurs in the normal course of cardiac surgery during aortic cross-clamping (AXCL), and plasma level of cardiac troponin, a specific myocardial injury marker, increases during reperfusion after releasing the clamp. Earlier clinical studies also found C3, the common factor in three complement pathways, is activated during cardiac surgery. In this prospective study, we investigated three complement pathways in the involvement of myocardial injury during cardiac surgery. Fifty patients undergoing open heart surgery were enrolled. Coronary sinus and peripheral blood were collected at different peri-operative time points. Plasma levels of the initial factors in the complement pathways were analyzed. Our results showed that the level of complement factor B, the initial component in the alternative pathway, increased significantly in both coronary sinus and peripheral blood after release of AXCL compared to samples taken prior to AXCL. Spearman correlation analysis showed that this post-AXCL increase in factor B was significantly correlated with the cardiac troponin level sampled immediately after the operation. In contrast, there was no significant correlation between the initial factors in the classical or the lectin complement pathways and post-operation troponin levels. Thus, alternative complement pathway is activated during open heart surgery and may play a role in human myocardial ischemic injury.
NKG2D/ligands-mediated immune activation promotes atherosclerosis (142.7)
Authors: Xia Mingcan , Yang Kangkang , Guerra Nadia , Sukhova Glina , Miller Carla , Shi Guo-Ping , Raulet David , Xiong Na 
Published: January 2023

Abstract
Atherosclerosis, a chronic vascular disease, is characterized by deposition of fatty substances and accumulation of immune cells in susceptible regions of arteries. Increasing evidence show that abnormal metabolic conditions, such as dyslipdemia and diabetes, induce immune cell activation and inflammation, which play crucial roles in progression of atherosclerosis. However, molecular linkers of metabolic disorder, immune activation and atherosclerosis are not well understood. Here we report that ligands for NKG2D, an immune-activating receptor, were upregulated in multiple tissues and organs under atherosclerotic disease condition, particularly in atherosclerotic lesion and liver. Preventing NKG2D/ligands interaction significantly reduced atherosclerotic lesion formation and systemic inflammation. In addition, preventing NKG2D/ligand interaction alleviated abnormal metabolic conditions by suppressing liver inflammation. Our data clearly show that NKG2D/ligand interaction plays critical roles in the progression of atherosclerosis and might be potential targets against atherosclerosis.
Regulation of the Receptor Activator of NF-κB Ligand (RANKL)-induced activation of the alternative NF-κB pathways by interleukin-4 (IL-4) (142.8)
Authors: Yu Minjun , Moreno Jose , Keegan Achsah 
Published: January 2023

Abstract
NF-κB signaling is essential for RANKL-induced osteoclast (OC) formation. IL-4 inhibited RANKL-induced OC differentiation, while at the same time promoted macrophage fusion to form multinucleated giant cells (MNG). Several groups have proposed that IL-4 acted by suppressing the RANKL-induced activation of NF-κB. However, we found that IL-4 did not block canonical NF-κB signaling. Instead, we found that IL-4 inhibited alternative NF-κB signaling and induced p105/50 expression. Interestingly, in p105/50 deficient bone marrow macrophages (BMM), the formation of both multinucleated OC and MNG induced by RANKL or IL-4 respectively was impaired. This suggests that NF-κB signaling also plays an important role during macrophage fusion and MNG formation. To confirm this, the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) was used; PDTC blocked both RANKL-induced OC and IL-4-induced MNG formation. RT-PCR and western blot analyses showed that E-cadherin, and DC-STAMP, proteins important for macrophage fusion, were downregulated by PDTC. Furthermore, overexpression of p52 or RelB in p105/50 deficient BMM significantly enhanced both OC and MNG formation. DC-STAMP was upregulated in p52 and RelB transduced BMMs. These results suggest that the influence of IL-4 on NF-κB activation pathways is complex, and that NF-κB pathways positively regulate the IL-4-induced formation of MNG by MCSF-dependent macrophages.