bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mast cells are initiators of inflammatory pain in thermal pain in mice. (86.2)
Authors: Chatterjea Devavani , Engblom Camilla , Mora-Solano Carolina , Maitland-McKinley Paul , Allen Juliann 
Published: January 2023

Abstract
Pain is a cardinal component of inflammation and a significant public health issue. Cytokines such as TNF-alpha, IL-1beta, IL-15 and CXCL1/IL-8, as well as neutrophil influx have been implicated in the potentiation of peripheral inflammatory pain in models of thermal and mechanical hindpaw hypernociception in rodents. These lines of evidence point to sentinel mast cells in the tissues as possible regulators of these pain responses. Degranulation of mast cells has been shown to be important for phosphorylation of Erk in nociceptive neurons in a rat model of migraine-like stimulation. Mechanical bladder pain in a model of interstitial cystitis was abrogated in mast cell deficient mice. We show that c4880-mediated degranulation of mast cells causes pain as well as significant edema and upregulation of local TNF-alpha and IL-1beta levels in the hindpaws of mice in a model of thermal pain. Mast cell-deficient Wsh mice show abrogation of this pain response. IgE/Ag mediated degranulation of mast cells in the paws of WT mice also causes pain and edema but it is not accompanied by detectable upregulation of TNF-alpha in the paws. This suggests that mast cells may be necessary for the initiation of inflammatory pain but that the underlying mechanisms of potentially protective inflammatory pain vs. potentially pathological allergic pain may be different.
Biomarkers of IgE- and IgG-dependent anaphylaxis (86.20)
Authors: Khodoun Marat , Strait Richard , Armstrong Laura , Yanase Noriko , Perkins Charles , Potter Crystal , Herbert De’Broski , Finkelman Fred 
Published: January 2023

Abstract
Anaphylaxis is classically induced by IgE-dependent FceRI crosslinking that triggers mast cell and basophil mediator, enzyme and cytokine secretion. Anaphylaxis can also be induced in mice, however, by IgG- mediated crosslinking of macrophage and basophil FcgRIII. This alternative pathway may be responsible for some cases of human anaphylaxis, particularly those associated with protein-based drugs, including humanized mAbs. We used mouse passive anaphylaxis models to attempt to identify markers that could distinguish between IgE- and IgG-mediated anaphylaxis. Mice sensitized by i.v. injection of TNP-specific IgE or IgG mAbs were challenged i.v. 24h later with TNP-OVA; changes in cells and serum were then evaluated. Changes 2h post-challenge in both IgE- and IgG-mediated anaphylaxis included: 1) up to 90% decreases in percentages of basophils and monocytes in blood; 2) doubling of the percentage of neutrophils in blood; and 3) ~25% increases in the ability of neutrophils to generate reactive oxygen intermediates and mobilize Ca++. IgE-, but not IgG-mediated anaphylaxis induced greatly increased serum levels of IL-4 and MMCP1 and an ~30% increase in T cell membrane IL-4Ralpha 4 h post-challenge, while IgG-, but not IgE-dependent anaphylaxis decreased neutrophil membrane FcgRIIb/RIII expression by ~30% 2-12 h post-challenge. Studies will be performed to determine if these cellular markers can distinguish between IgE-dependent and putative IgG-dependent anaphylaxis in humans.
Novel G-Protein Coupled Receptors in the Activation of Human Mast Cells by C3a-Related Peptides (86.23)
Authors: Kashem Sakeen , Subramanian Hariharan , Magotti Paola , Lambris John , Ali Hydar 
Published: January 2023

Abstract
Studies with animal models have shown that complement components C3a and C5a play an important role in the effector phase of allergic asthma. We have previously shown that C3a activates human CD34+-derived mast cells endogenously expressing G protein coupled receptors (GPCR) for C3a (C3aR) as well as RBL-2H3 cells stably expressing C3aR. The purpose of this study was to utilize peptide C3a receptor agonists to further characterize the role of C3aR on mast cell activation. We found that C3a (a 74 amino acid peptide) and a C3aR peptide superagonist (H-WWGKKYRASKLGLAR-OH) activated human mast cells with similar potency. Surprisingly, a scrambled C3aR superagonist peptide also caused mast cell degranulation. RT-PCR analysis demonstrated that all four known human Mas-related genes, MrgX1, MrgX2, MrgX3, and MrgX4 are expressed in human mast cells. RBL-2H3 cells stably expressing MrgX2 responded to its known ligand, cortistatin-14 for sustained Ca2+ mobilization and degranulation. Furthermore, C3aR superagonist peptide, scrambled C3aR superagonist peptide and substance P induced degranulation in RBL-2H3 cells expressing MrgX2 but not in untransfected cells. These findings suggest that Mrg receptors may play a novel role in activation of mast cell by complement-derived and other positively charged peptides and could serve as therapeutic targets for mast cell-dependent allergic and inflammatory diseases.
TGFb1 and IL-4 Provide Antagonistic Control of the Mast Cell Response (86.5)
Authors: Ryan John , Morales Johanna , Macey Matthew 
Published: January 2023

Abstract
Cytokine networking is an important concept for understanding how the immune system is controlled. We hypothesized that the initial presence of pro-inflammatory cytokines could desensitize mast cells to anti-inflammatory signals, rendering them more likely to be activated in allergic reactions. Conversely, the initial presence of anti-inflammatory cytokines might suppress the effects of pro-inflammatory cytokines in allergic type responses. In this study, we show that IL-4 and TGFβ1 act as an antagonistic pair in controlling the mast cell response. Mast cell pre-culture with IL-4 decreased TGFβR1 and TGFβR2 RNA and protein levels through a STAT6-dependent mechanism. Further, IL-4 diminished TGFβ1 signaling, blocking SMAD2 phosphorylation. IL-4 also blocked TGFβ1-mediated mast cell death and suppressed TGFβ1-mediated mast cell migration. Pre-culture with TGFβ1 mirrored this antagonistic effect. TGFβ1 pre-culture reduced IL-4R protein levels, suppressed IL-4-mediated Stat6 phosphorylation, and diminished mast cell migration toward IL-4. When both are present simultaneously, TGFβ1 appears to have a dominating inhibitory effect over IL-4 by blocking proliferation in vitro and macrophage recruitment in vivo. We obtained similar results using human mast cells. These data are important to understanding how cytokine networks can impact mast-cell mediated diseases like asthma. Supported by NIH grants 1R01 AI59638 and U19A1077435.
Eosinophils regulate immunity during chronic infection by Trichinella spiralis (87.14)
Authors: Gebreselassie Nebiat , Fabre Valeria , Bliss Susan , Gagliardo Lucille , Lee Nancy , Lee James , Appleton Judith 
Published: January 2023

Abstract
Eosinophilia is a hallmark of worm infection and allergy. The parasitic nematode Trichinella spiralis establishes chronic infection in skeletal muscle. We have shown previously that in eosinophil-ablated, transgenic mice, T. spiralis larvae die in large numbers in the muscle. Death is promoted by inducible nitric oxide synthase (iNOS). We report here that transcription of iNOS was upregulated in the context of an established alternative activation phenotype in local leukocytes. Infiltrating macrophages, dendritic cells, and neutrophils produced iNOS. These changes were preceded by a reduced T cell recruitment to infected muscle that was coincident with a lag in larval growth, suggesting compromise of the parasite’s habitat. The results further define the cellular interactions that occur at a site of chronic nematode infection in which the eosinophil functions as a pivotal regulator of immunity.
Parthenolide, an anti-inflammatory NFκB inhibitor, stimulates platelet production and blunts platelet activation (87.17)
Authors: Sahler Julie , Bernard Jamie , Spinelli Sherry , Casey Ann , Blumberg Neil , Phipps Richard 
Published: January 2023

Abstract
Platelets play a significant role in hemostasis, and are critically involved in common medical complications including thrombocytopenia, ischemia, and atherosclerosis. Platelets can also directly induce inflammation and influence immune cells by releasing inflammatory mediators such as CD40 Ligand, TGF-β, PGE2, IL-1β, and MIP-1α. Despite these recent findings, platelet signaling pathways remain unclear and are under current investigation. Recently, our lab identified the presence of NFκB transcription factors in platelets. Because platelets are anucleate, we hypothesize an alternative role for NFκB in platelet function. To further study the role of NFκB in platelets, we tested the effects of an NFκB inhibitor, Parthenolide, on human platelets and Meg-01 cells, a human megakaryoblastic cell line. Parthenolide is an extract of the Feverfew plant used in herbal therapies to dampen inflammation. We found that Parthenolide directly influenced human platelets by diminishing clot retraction and spreading. In addition, we found that Parthenolide increased platelet production, increased heme oxygenase-1 (up to 2000-fold), and increased glutathione levels over time (p=0.02) in Meg-01 cells. These data suggest that the anti-inflammatory properties of Parthenolide may be partially due to its ability to suppress platelet activation and up-regulate anti-oxidant pathways.
Ptaquiloside-induced immunotoxic effects are reversed by selenium supplementation (87.21)
Authors: Latorre Andreia , Caniceiro Beatriz , Wysocki Harry , Haraguchi Mitsue , Gardner Dale , Górniak Silvana 
Published: January 2023

Abstract
Pteridium aquilinum (Pa) is a poisonous plant and its ingestion has been closely associated with human and animal cancers. Our earlier studies have shown that Pa reduces white pulp of spleen and NK cell cytotoxicity in mice. In addition, we observed that selenium (Se) reversed Pa-induced spleen damage. In this study the aim was evaluate if ptaquiloside (Ptq), the main toxic principle found in Pa, is responsible for its immunotoxic effects and if Se would reverse these effects. For in vivo study, mice were separated into 4 groups: control, Ptq, Ptq+Se, and Se. Mice were treated by gavage for 14 days with Ptq (5.3mg/kg BW) and/or Se (1.3mg/kg BW). Mice were killed and it was prepared a non-adherent splenic cells suspensions to perform NK cell activity assay against YAC cell line. For in vitro study, cultures of non-adherent splenic cells were performed for each mouse as follows: untreated, Se 0.1mM, RPMI extract of Pa (RE) 4mg/ml, RE 4mg/ml+Se 0.1mM (co-incubation), and RE 4mg/ml+Se 0.1mM (RE/1h wash and then Se/1h). After treatment, the cells were washed to proceed both NK cell activity assay and MTT assay. We observed that Ptq reduced NK cytotoxicity, which permits to suppose that Ptq is the principle responsible by immunotoxic effects. Moreover, NK cell activity was completely recovered after Se supplementation in vivo and in vitro. These results showed for the first time that Pa-induced immunotoxic effects are caused by Ptq and that these effects are fully recovered by Se.
Involvement of estrogen receptor alpha on MCP1 production in C57BL/6 kidney mesangial cells (87.29)
Authors: Dasgupta Subhajit , Eudaly Jackie , Molano Ivan , Baker DeAnna , Mollenhauer Meagan , Gilkeson Gary 
Published: January 2023

Abstract
Abstract We demonstrate an estrogen independent involvement of ERα in TLR2 mediated expression of MCP1 in mesangial cells. In vitro treatment with the TLR2 ligands Pam3CysSK4, LTA and PGN-SA induced production of MCP1 in mesangial cells isolated from kidneys of female C57BL/6 mice in a dose dependent manner. Treatment with LTA to C57BL/6 mesangial cells was found to activate NF kB and induced its nuclear localization. Interestingly enough, we found LTA treatment induced phosphorylation of ERα in primary mesangial cells and also in SV40 virus transformed B6 mesangial cell line. Nuclear translocation of pERα (Ser118) was found in the LTA treated B6 mesangial cells. ERα selective antagonist MPP and NF kB antagonist TPCA1 attenuated MCP1 production in TLR2 ligand treated mesangial cells suggesting influence of pERα in NF kB dependent MCP1 gene expression. The LTA was found to induce DNA binding activity of NF kB in B6 mesangial cell line in a time dependent manner. Transient transfection with ERα siRNA was also found to reduce expression of MCP1 in Pam3CysSK4 treated mesangial cells. Taken together, our study indicates an unique role of ERα in modulation of TLR2-mediated signals in mesangial cells. Acknowledgement: We are thankful to the Department of Medicine, Division of Rheumatology/Immunology and Ralph V Johnson VA research service, MUSC. The research support was provided by VA research service. We do not have any conflict of interests.
IL-17 plays differential role in primary verse memory immune responses and recognition of cellular verse soluble antigens (87.30)
Authors: Guan Hongbing , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
IL-17 is a proinflammatory cytokine that has been linked to many autoimmune related diseases such as rheumatoid arthritis, asthma, lupus, and multiple sclerosis. Recent studies have indicated that IL-17 may also play central role in driving the development of germinal center-derived autoantibody and Th response in an autoimmune disease model setting. In addition, activated and memory T cells can both produce IL-17. In this report, we found that IL-17 differentially affected primary versus memory immune response to cellular versus soluble antigen. Briefly, we initiated primary or memory immune response with SRBC or OVA and blocked IL-17 using anti-IL-17 antibody (Amgen). We found that blockade of IL-17 did not affect OVA-induced primary or memory response, nor did it affect SRBC-induced memory response; however, the blockade inhibited SRBC-induced primary response. SRBC-induced hemagglutination, DTH and specific T cell proliferation were inhibited; serum IFN-gamma, IL-12 and SRBC-specific IgG1 were decreased; numbers of germinal centers and IgG1-producing B cells in the germinal center were decreased; IL-17R knockout-CD4 T cells showed a decreased expression of BTLA and ICOS; however, percentages of CD4, CD8, CD3 and CD19 subsets in the spleen did not change with the antibody blockade. Our report suggests a differential role of IL-17 on T-B, T-DC cell interaction or Ig class switch in response to cellular versus soluble antigens, and primary versus memory immune response.
Complement Factor H regulation in the central nervous system (87.5)
Authors: Fraczek Laura , Martin Carol , Martin Brian 
Published: January 2023

Abstract
The complement system is a critical component of innate immunity that requires regulation to avoid inappropriate activation. This regulation is provided by many proteins including complement Factor H (CFH), a secreted protein of the alternative pathway. Despite its regulatory function, gene and promoter polymorphisms of CFH have been implicated in patients with age-related macular degeneration and atypical hemolytic uremic syndrome, indicating that a better understanding of this gene’s regulation is valuable. Presently, little work has been done to address the role of CFH transcriptional control in the central nervous system (CNS). We have cloned the murine CFH promoter and have made truncation constructs to examine CFH gene regulation in cells of the CNS. Our studies show that CFH mRNA is present in several CNS cell types, and that specific regions throughout the promoter contain enhancers and repressors that are positively and negatively regulated by inflammatory cytokines. Database mining of these regions indicates that there are potential transcription factor binding sites conserved between different species, which led us to investigate specific transcription factor binding interactions in these regions. Through these studies we hope to elucidate the transcriptional regulation of CFH in the CNS.
Regulation of CD20 in rituximab-resistant cell lines (RRCL) and B-cell non-Hodgkin’s lymphomas (NHL) (88.18)
Authors: Tsai Ping-Chiao , Naveen Bangia , Olejniczak Scott , Hernandez-Ilizaliturri Francisco , Czuczman Myron 
Published: January 2023

Abstract
Rituximab(R) resistance is associated with low CD20 expression in relapsed/refractory (r/r) lymphomas. Studies had shown a direct correlation between CD20 levels and R-induced complement cell lysis. CD20 up-regulation by IL-4 serves as a useful tool to study regulatory mechanisms of CD20 expression. We demonstrated that chronic exposure of B-cell to R leads to decrease CD20 at mRNA and protein levels. Our current work defines regulatory mechanisms of CD20 expression in RRCL and evaluates the role of CD20 in R activity. Differences in CD20 mRNA stability and promoter activity were determined between RSCL and RRCL by Northern blot and luciferase reporter assays. Levels of CD20 regulatory factors were determined by Western Blot. IL-4 effects in CD20 promoter activity, CD20 surface expression and R activity was studied in RSCL, RRCL and in tumor cells derived from r/r NHL. No difference in CD20 mRNA stability was noted between RSCL and RRCL. CD20 promoter activity was lower in RRCL. Detailed analysis of different CD20 promoter fragments indicated that low CD20 promoter activity occurred within the first 300 base pairs, suggesting lack of positive regulatory factors. Exposure to IL-4 increased CD20 promoter activity, CD20 expression and R activity. Our data suggest that CD20 promoter activity plays a role in CD20 down-regulation in RRCL. We are currently studying differences in PU.1 or Oct-1 binding to CD20 promoter in an attempt to define their role, if any, in R-resistance
Innate production of Th2 cytokines by adipose tissue-associated c-Kit+Sca-1+ lymphoid cells (89.11)
Authors: Moro Kazuyo , Koyasu Shigeo 
Published: January 2023

Abstract
Innate immune responses are important in combating various microbes during the early phases of infection. NK cells are innate lymphocytes that, unlike T and B lymphocytes, do not express antigen receptors but rapidly exhibit cytotoxic activities against virus-infected cells and produce various cytokines. Here we report a new type of innate lymphocyte present in a novel lymphoid structure associated with adipose tissues in the peritoneal cavity. These cells do not express lineage (Lin) markers but do express c-Kit, Sca-1, IL7R and IL33R. Similar lymphoid clusters were found in both human and mouse mesentery and we term this tissue ‘FALC’ (fat-associated lymphoid cluster). FALC Lin-c-Kit+Sca-1+ cells are distinct from lymphoid progenitors and lymphoid tissue inducer cells. These cells proliferate in response to IL2 and produce large amounts of Th2 cytokines such as IL5, IL6 and IL13. IL5 and IL6 regulate B-cell antibody production and self-renewal of B1 cells. Indeed, FALC Lin-c-Kit+Sca-1+ cells support the self-renewal of B1 cells and enhance IgA production. After helminth infection and in response to IL33, FALC Lin-c-Kit+Sca-1+ cells produce large amounts of IL13, which leads to goblet cell hyperplasia—a critical step for helminth expulsion. In mice devoid of FALC Lin-c-Kit+Sca-1+ cells, such goblet cell hyperplasia was not induced. Thus, FALC Lin-c-Kit+Sca-1+ cells are Th2-type innate lymphocytes, and we propose that these cells be called ‘natural helper cells’.
Synergistic Signals for Natural Cytotoxicity Are Required to Overcome Inhibition by c-Cbl (89.16)
Authors: Kim Hun Sik , Das Asmita , Gross Catharina , Bryceson Yenan , Long Eric 
Published: January 2023

Abstract
Natural killer (NK) cells do not express variable antigen-specific receptors but are activated by combinations of germ-line encoded receptors. How activation is achieved and controlled during encounter of NK cells with target cells is unclear. We showed here that co-engagement of receptors NKG2D and 2B4 triggered synergistic activation of PLC-γ2, degranulation and cytotoxicity, which were controlled positively by Vav1 and negatively by the E3 ubiquitin ligase c-Cbl. Knockdown of c-Cbl blocked ubiquitylation of Vav1 and bypassed the requirement for synergy, as NKG2D and 2B4 could induce degranulation on their own. The enhanced degranulation obtained after c-Cbl knockdown was still Vav1-dependent. Inhibition of cytotoxicity through binding of CD94-NKG2A to HLA-E on target cells was independent of inhibition by c-Cbl. Therefore, natural cytotoxicity is controlled not only by dominant inhibitory receptors, but also by an activation threshold set by c-Cbl, which is overcome by Vav1-dependent synergy of coactivation receptors.
Calcium hydroxide attenuates the inflammatory potential of lipoteichoic acid (LTA) from Enterococcus faecalis by deacylating the LTA (89.22)
Authors: Baik JungEun , Yun Cheol-Heui , Han SeungHyun 
Published: January 2023

Abstract
Enterococcus faecalis is a pathogenic gram-positive bacterium closely related with refractory apical periodontitis. Since lipoteichoic acid (LTA) is a major virulence factor of gram-positive bacteria, we prepared highly pure LTA from E. faecalis and investigated its inflammatory responses in a murine macrophage-like cell line, RAW 264.7. Upon exposure to the E. faecalis LTA, the cells substantially produced tumor necrosis factor-α (TNF-α) and nitric oxide (NO) in a dose- and time-dependent manner. Notably, the action of the LTA was through Toll-like receptor 2 (TLR2), but not TLR4. The LTA enhanced the DNA-binding activity of a transcription factor, NF-κB, which plays an important role in the transcriptional activation of genes encoding various inflammatory mediators. However, induction of TNF-α and NO production was not observed in the cells exposed to LTA pre-treated with calcium hydroxide, which is a widely used as intracanal medicament for sterilizing root canals of teeth. Furthermore, calcium hydroxide-treated LTA was not able to stimulate TLR2. Structural analysis using MALDI-TOF mass spectrometry showed that calcium hydroxide deacylated the glycolipid moiety in the LTA structure. Conclusively, we suggest that calcium hydroxide attenuates the inflammatory activity of E. faecalis LTA through deacylation of the LTA structure.
Crucial role of calcium signaling in TCR-dependent functions of Vgamma2Vdelta2 T cell (89.23)
Authors: Li Haishan , Pauza C. David 
Published: January 2023

Abstract
Vgamma2Vdelta2 T cells in human peripheral blood recognize phosphoantigen and play important roles in host defense and immunoregulation. However, the molecular mechanisms responsible for phophogantigen-mediated activation of human gammadelta T-cells remain unclear. We have previously reported that phosphoantigen isopentyl pyrophosphate (IPP) activated MEK/Erk and PI-3K/Akt pathways rapidly. Specific inhibition of Erk and Akt significantly impaired the functional response to IPP. In the present study, we found that IL2 can also activate MEK/Erk and PI-3K/Akt pathways without inducing functional responses. When given together with IPP, the effect of IL2 was to enhance cytokines production, degranulation and cytotoxicity. These data implicate the activation of MEK/Erk and PI-3K/Akt pathways as necessary but not sufficient to induce functional responses in Vgamma2Vdelta2 T cells. Further, we found that FK506 or cyclosporine A (CsA) inhibitors of calcineurin, completely blocked IPP-induced cytokines production, and partly inhibited TCR-dependent degranulation and cytotoxicity functions of Vgamma2Vdelta2 T cells. IPP but not IL2 induced the calcium-dependent nuclear translocation of NFAT. Our results indicate that calcium signaling pathway plays a crucial role in TCR-dependent functions in Vgamma2Vdelta2 T cells that can be distinguished from the effect of IL2.