bluuebunny/crossref_metadata_2025_split huggingface dataset:
The −180 Site of the IL-2 Promoter Is the Target of CREB/CREM Binding in T Cell Anergy
Authors: Powell Jonathan D. , Lerner Cara G. , Ewoldt Gerald R. , Schwartz Ronald H. 
Published: December 2022

AbstractAnergic T cells display a marked decrease in their ability to produce IL-2 even in the presence of optimal TCR and costimulatory signals. Using IL-2 enhancer/promoter-driven reporter constructs, we have previously identified a region that appears to be a target for cis transcriptional repression in anergy. This region of the promoter, which shares partial homology with a consensus AP-1-binding sequence, is located about −180 bp from the transcriptional start site. In the present study, we demonstrate that cAMP response element-binding protein/cAMP response element modulator (CREB/CREM), activating transcription factor-2/c-Jun, and Jun-Jun/Oct complexes bind to this site. However, the induction of anergy by prolonged stimulation through the TCR led to an increase in binding of only the CREB/CREM complex. Furthermore, the level of binding of this complex appeared to be up-regulated in both resting and restimulated anergic T cells. Finally, an IL-2 promoter-driven reporter construct that contained a mutation that specifically reduced the binding of the CREB/CREM complex displayed a decreased ability to be affected by anergy, while a construct that contained a mutation that decreased the binding of the Jun-Jun/Oct complex was still susceptible to anergy. These findings suggest that the −180 region of the IL-2 promoter is the target of a CREB/CREM transcriptional inhibitor that contributes to the repression of IL-2 production in T cell anergy.
Contributions of Leukemia Inhibitory Factor Receptor and Oncostatin M Receptor to Signal Transduction in Heterodimeric Complexes with Glycoprotein 130
Authors: Hermanns Heike M. , Radtke Simone , Haan Claude , Schmitz-Van de Leur Hildegard , Tavernier Jan , Heinrich Peter C. , Behrmann Iris 
Published: December 2022

AbstractLeukemia inhibitory factor (LIF), cardiotrophin-1, ciliary neurotrophic factor, and oncostatin M (OSM) lead to heterodimerization of LIF receptor (LIFR) or the OSM-specific receptor (OSMR) with glycoprotein (gp) 130, the common receptor subunit for IL-6-type cytokines. Thereby intracellular signaling via Janus kinases (Jaks) and STAT transcription factors is initiated. We investigated the contributions of LIFR and OSMR to signal transduction in the context of heterodimers with gp130. Chimeric receptors based on the extracellular parts of the IL-5R α- and β-chains were generated, allowing the induced heterodimerization of two different cytoplasmic tails. Our studies demonstrate that upon heterodimerization with the gp130 cytoplasmic region, the cytoplasmic parts of both LIFR and OSMR were critical for activation of an acute phase protein promoter in HepG2 hepatoma cells. The membrane-proximal region of LIFR or OSMR was crucial for the ability of such receptor complexes to induce DNA binding of STAT1 and STAT3 in COS-7 cells. Membrane-distal regions of LIFR and OSMR contributed to STAT activation even in the absence of gp130 STAT recruitment sites. We further show that the Janus kinases Jak1 and Jak2 constitutively associated with receptor constructs containing the cytoplasmic part of LIFR, OSMR, or gp130, respectively. Homodimers of the LIFR or OSMR cytoplasmic regions did not elicit responses in COS-7 cells but did in HepG2 cells and in MCF-7 breast carcinoma cells. Thus, in spite of extensive functional similarities, differential signaling abilities of gp130, LIFR, and OSMR may become evident in a cell-type-specific manner.
Nitric Oxide Plays a Critical Role in the Recovery of Lewis Rats from Experimental Autoimmune Encephalomyelitis and the Maintenance of Resistance to Reinduction
Authors: O’Brien Nikki C. , Charlton Brett , Cowden William B. , Willenborg David O. 
Published: December 2022

AbstractExperimental autoimmune encephalomyelitis (EAE) is a T cell-mediated autoimmune disease of the CNS and an animal model for the human demyelinating disease, multiple sclerosis. In the Lewis rat, myelin basic protein (MBP)-CFA-induced EAE is an acute monophasic disease from which animals recover fully, do not relapse, and develop a robust long-term resistance to further active reinduction of disease. In this paper, we report that rats recovering from MBP-CFA-induced EAE have significantly increased serum levels of reactive nitrogen intermediates indicative of increased NO production. These levels remain elevated after the recovery period and increase even further early after a rechallenge with MBP-CFA, and all animals are totally refractory to a second episode of disease. Oral treatment of rats with N-methyl-l-arginine acetate (l-NMA), beginning at peak disease on day 11 postimmunization, results in significant prolongation of disease and an alteration in the presentation of clinical symptoms from that of solely hind limb paresis/paralysis to severe fore limb involvement as well. Treatment of fully recovered rats with l-NMA 24 h before a rechallenge with MBP-CFA leads to decreased serum reactive nitrogen intermediate levels and results in a second episode of EAE in 100% of animals. Furthermore, l-NMA treatment of fully recovered rats in the absence of a rechallenge immunization leads to spontaneous relapse of disease.
The soy peptide lunasin enhances cross-presentation by dendritic cells (APP3P.109)
Authors: Tung Chun-Yu , Casiano-Rivera Félix , Yao Shuyu , Sun Jie , Chang Hua-Chen 
Published: December 2022

Abstract
Lunasin, a natural seed peptide containing 43 amino acids, was originally isolated from soybeans. We recently discovered a novel function of lunasin as an immune modulating agent. Lunasin stimulation resulted in dendritic cell (DC) maturation and activation by increasing the expression of co-stimulatory molecules and the production of type I interferons, pro-inflammatory cytokines and chemokines. Lunasin acts as an adjuvant in vivo was demonstrated in a prophylactic model against OVA-expressing A20 B-lymphomas. The objective is to determine the mechanisms by which lunasin promotes antigen-specific antitumor immune responses. We hypothesize that lunasin enhances cross-presentation of tumor antigens by DCs, resulting in priming of tumor specific cytotoxic T lymphocytes (CTLs). To test our hypothesis, in vitro cross-presentation of soluble OVA by isolated DCs (CD11c+) was performed. Lunasin-treated DCs stimulated greater proliferation of OVA specific CD8+ T cells (OT-I) cells than DCs cultured in medium only. To further define the molecular mechanisms, we determined the activation of transcription factors involved in DC maturation by lunasin. Increases in phosphorylation of STAT1 and NF-kB were found in DCs treated with lunasin. The data may delineate the signaling pathways involved in gene expression induced by lunasin in DCs. These results show that lunasin enhances cross-presentation of tumor antigens by DCs, resulting in effective priming of antigen specific CD8+ CTLs.
Macrophage anti-viral responses to diabetogenic Coxsackie virus infections are mediated by free radicals (BA6P.134)
Authors: Das Shaonli , Anderson Brian , Burg Ashley , Tse Hubert 
Published: December 2022

Abstract
Reactive oxygen species (ROS) induces Type-1 diabetes (T1D) by directly killing pancreatic β-cells, triggering pro-inflammatory cytokine production, and promoting autoreactive T cell maturation. We hypothesize that free radical synthesis in T1D can synergize and enhance autoimmune responses in T1D onset and progression. Free radicals can be identified by immuno-spin trapping with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and utilizing a DMPO-specific antibody to detect DMPO-radical adducts. We previously reported a delay in spontaneous T1D onset in Non-obese diabetic (NOD) mice impaired in NADPH Oxidase (NOX)-derived superoxide synthesis (NOD.Ncf1m1J). Bone marrow-derived macrophages (BM-Mϕ) from NOD.Ncf1m1J mice infected with Coxsackie virus CB3 strain displayed a significant decrease (p&lt;0.0001) in fluorescence intensity of DMPO adducts at 4, 6,12 and 24 hours post-infection concomitant with a decrease (p=0.0027) in STAT1 phosphorylation compared to NOD BM-Mϕ. CB3 infected-NOD.Ncf1m1J BM-Mϕ also exhibited a decrease in Tnfa (p=0.0037) and Ifih1 (p=0.002) mRNA accumulation in contrast with NOD BM-Mϕ. These findings suggest that anti-viral responses in NOD.Ncf1m1J BM-Mϕ are redox-regulated and were compromised by ROS-deficiency. Our results further illustrate the importance of NOX-derived superoxide in modulating autoimmune responses in T1D. Future studies will utilize immuno-spin trapping to visualize the interface between free radicals and immune cells during T1D progression.
Understanding neuronal gene expression changes induced by distal demyelination or inflammatory insult (BA6P.137)
Authors: Clarkson Ben , Mirchia Kanish , Kim Hyo , Sauer Brian , LaFrance-Corey Reghann , Howe Charles 
Published: December 2022

Abstract
MR imaging suggests an anatomic correspondence between WM and GM lesions in progressing MS patients, suggesting an underlying neuronal-signaling or neurodegenerative element to the progressive disease state. While retrograde signals evolving from stressed axons that alter soma gene expression have been previously described in models of axonal transection or crush injury, those resulting from inflammation and demyelination are unknown. Using microfluidic chambers that segregate neuronal soma and axons, we have previously demonstrated increased axonal MHC I following treatment with IFNg, a cytokine prominent in demyelinating MS lesions. Furthermore, expression of antigen-loaded MHC I predisposed axons to injury by antigen-specific CD8+ T cells in vitro. Here we report that neurons also upregulate expression of MHC class Ib mRNAs in response to axonal stimulation with IFNg. Genes most highly upregulated following cytokine treatment included H2-M3, H2-Q5, H2-Q6, H2-Q7, H2-Q8, H2-T10 , and H2-T23. We further report the development of an in vivo model system for measuring axonal vulnerability to antigen-specific immune attack following demyelination using an AAV vector to induce neuron-specific expression of axonally-targeted neoantigen (ovalbumin) tagged with cleavable eGFP. We also present optimized methods using AAV vectors in mice for infecting retinal, motor cortex, and hippocampal neurons to induce target gene expression in axons projecting to optic nerve and corpus collosum.
Neutrophils regulate humoral autoimmunity by restricting interferon gamma production via the generation of reactive oxygen species (BA7P.142)
Authors: Cao Wei , Li Jingjing , Huang Xinfang , Dorta-Estremera Stephanie , Di Domizio Jeremy , Anthony Scott , Schluns Kimberly , Lanier Lewis 
Published: December 2022

Abstract
The mechanism by which plasmacytoid dendritic cells (pDCs) promote humoral autoimmunity is unclear. In amyloid-induced experimental autoimmune model, pDC activation is accompanied by prominent neutrophilia. We investigated the contribution of neutrophils since neutrophils have been critically implicated in the pathogenesis of systemic autoimmunity. Unexpectedly, neutrophil depletion enhanced anti-nuclear antibody development, which was correlated with heightened IFN-γ production by natural killer (NK) cells. IFN-α/β produced by pDCs activated NK cells via the induction of IL-15. Neutrophils released reactive oxygen species (ROS), which negatively modulated the levels of IL-15, thereby inhibiting IFN-γ production. Consistently, mice deficient of NADPH oxidase 2 (Nox2) produced increased amounts of IFN-γ and developed augmented titers of autoantibodies. Both the pDC-IFN-α/β pathway and IFN-γ were indispensable in stimulating humoral autoimmunity. Male NZB/W mice had higher levels of superoxide than their female lupus-prone siblings and depletion of neutrophils resulted in spontaneous NK cell and autoimmune B cell activation. Our findings reveal a regulatory role of neutrophils in vivo and highlight the importance of IFN-γ downstream of the pDC-IFN-α/β pathway in systemic autoimmunity.
Role of dendritic cells in immunopathogenesis of Pemphigus vulgarism (BA7P.149)
Authors: Antil Parul , Das Dayasagar , Khandpur Sujay , Sharma Vinod , Singh Manoj , Sharma Alpana 
Published: December 2022

Abstract
Pemphigus vulgaris (PV) is an autoimmune blistering disease that affects skin and mucous membranes characterized by autoantibodies against Dsg3. The breach in immune tolerance, aberrant T cell function and antigen presentation are key immunological events in autoimmunity. Given the central role in induction of immune response and tolerance, dendritic cells (DCs) are most important cells in inciting and sustaining autoimmunity, but it has not yet been explored in PV. We have previously identified potential role of Th cells in pathogenesis of PV. In this study, DC frequency was estimated using HLA-DR, Lin1, CD11c and CD123 by flowcytometry. A significant decline in mDC & pDC frequency in circulation of PV patients (n=20) vs. controls (n=15) was observed. Further, molecular functional characteristics were assessed in cultured monocyte derived differentiated DCs and sorted DCs (FACS AriaIII) directly from circulation of patients and controls. The mRNA levels of their co-stimulatory molecules were estimated. Expression levels of stimulatory molecules (CD40 & CD80) was found to be significantly increased, while a significant decline in inhibitory markers (PSGL1 & ILT3) was observed in sorted and cultured DCs from patients vs. controls. Increased expression level of stimulatory molecules and decreased level of inhibitory molecules further strengthens the defect in functional status of DC. Thus, these co-stimulatory molecules might be a promising avenue for novel therapeutics in PV.
Antibody-mediated regulation of the response to a protective cryptic epitope in rMOG-induced EAE (BA8P.166)
Authors: Riter Melissa , Almatrook Alaa , Rambottom Michael , Cross Anne , Lyons Jeri-Annette 
Published: December 2022

Abstract
Multiple sclerosis is a Th1/Th17-mediated autoimmune disease of the central nervous system, in which B cells are involved. Experimental autoimmune encephalomyelitis (EAE) is the main animal model for MS. Previously we showed that B cell-deficient (B-/-) mice were resistant to rMOG-induced EAE. Further investigation revealed response to a cryptic epitope (MOG61-85) by lymph node (LN) T cells from rMOG-immunized B-/- mice but not WT mice. We investigated the immune response to MOG61-85 and the role of antibody in altering this response. EAE was ameliorated in B-/- mice when coimmunized with MOG35-55 and MOG61-85 compared with MOG35-55 alone. B-/- mice immunized with both peptides and injected with rabbit serum primed to MOG35-55 and MOG61-85 developed more severe EAE with earlier onset than mice receiving preimmune rabbit serum. Purified IgG from rMOG primed WT mice added to cultured LN T cells from co-immunized B-/- mice decreased T cell proliferation to MOG61-85, indicating that antibody inhibited the T cell response to MOG61-85. ELISA of culture supernatants from rMOG-primed WT LN cells demonstrated presence of TGF-β but not IL6 in response to MOG61-85; both cytokines were present in response to MOG35-55. B cell-/- LN cells cultured with MOG61-85 vs. MOG35-55 showed similar percentages of CD4+CD25+FoxP3+ T cells, whereas CD8+CD25+ T cells were slightly increased in those cultured with MOG61-85. The characterization of this CD8+ population is still under investigation.
IL-27 is negatively regulated by PGE2 in bone marrow-derived dendritic cells and macrophages (CCR4P.205)
Authors: Hooper Kirsten , Yen Jui-Hung , Kong Weimin , Ganea Doina 
Published: December 2022

Abstract
Interleukin-27 (p28/EBI3) is expressed by activated antigen presenting cells in response to TLR ligands and pro-inflammatory cytokines. The immunosuppressive functions of IL-27 have previously been demonstrated to occur through the induction of type 1 regulatory T cells, inhibition of Th17 cell development, and anti-inflammatory effects in murine models of collagen-induced arthritis (CIA), inflammatory bowel disease (IBD), and experimental autoimmune encephalomyelitis. We previously showed the pro-inflammatory effects of PGE2 in CIA and IBD through stimulation of IL-23 production leading to development of Th17 cells. Here we report on a novel pro-inflammatory function of PGE2 through the inhibition of IL-27 production by LPS-stimulated and poly I:C-stimulated murine bone marrow-derived dendritic cells (BMDC) and macrophages. We determined the effect of PGE2 on LPS-stimulated BMDC was mediated through EP2/EP4 receptors and induction of cAMP. Previous studies detailed the involvement of NF-κB (c-Rel) and interferon regulatory factors in IL-27p28 gene activation. The current project examines the effect of PGE2 on c-Rel, IRF1 and IRF3 utilizing luciferase reporter assays, gene expression and protein analysis. These studies further characterize the inflammatory mechanisms of PGE2, and might identify novel therapeutic targets for treating inflammatory diseases.
Regulation of virus-specific natural killer cell memory by IL-10 and TGF-β (CCR4P.207)
Authors: Geiger Theresa , Beaulieu Aimee , Vivier Eric , Li Ming , Sun Joseph 
Published: December 2022

Abstract
Natural killer (NK) cells are critical for host control of viral infections and are rapidly activated by pro-inflammatory cytokines. These activating cytokines are thought to be counter-balanced by regulatory cytokines to prevent excessive cytotoxicity or inflammation. This study investigates the role of the immunosuppressive cytokines transforming growth factor (TGF)-β and interleukin (IL)-10 in NK cell development and function, particularly in the generation of effector and memory responses against viral infection. We compared TGF-β-insensitive NK cells (using TGF-βRIIfl/fl x NKp46iCre mice) to NK cells from littermate control mice directly, as well as in mixed bone marrow chimera and adoptive transfer settings. Although they exhibited no obvious defect at steady state, TGF-β-insensitive NK cells revealed greater activation but decreased memory cell formation during mouse cytomegalovirus (MCMV) infection. Conversely, IL-10R-deficient mice at steady state contained greater percentages of mature NK cells in all organs. Similar to TGF-β-insensitive NK cells, IL-10R-deficient NK cells were also defective in generation of memory following MCMV infection. These studies reveal a novel and important role for immunosuppressive cytokines in regulating the magnitude of the effector response and in the optimal generation of long-lived NK cells following viral infection.
Innate lymphoid cell development requires TOX-dependent generation of a common ILC progenitor (HEM5P.221)
Authors: Seehus Corey , Aliahmad Parinaz , de la Torre Brian , Iliev Iliyan , Spurka Lindsay , Funari Vincent , Kaye Jonathan 
Published: December 2022

Abstract
Subtypes of innate lymphoid cells (ILC), defined by effector function and transcription factor expression, have recently been identified in both mice and humans. ILC are critical in maintaining barrier homeostasis and orchestrating cross-talk between the innate and adaptive arms of the immune system. In the adult, ILC derive from common lymphoid progenitors in bone marrow by passing through multiple ILC-specific progenitor stages, although transcriptional regulation of the developmental pathways involved in this process remains poorly defined. Previously, we showed that the nuclear DNA-binding protein TOX is required for development of lymphoid tissue inducer cells, a type of ILC3 required for lymph node organogenesis, and conventional natural killer cells, a type of ILC1. Here, we demonstrate that the production of multiple ILC lineages requires TOX, as a result of TOX-dependent development of common ILC progenitors. In vitro and ex vivo studies indicate that this deficit is cell intrinsic. Furthermore, using low-input RNA sequencing, we obtained robust whole transcriptome data from wild-type and TOX-deficient ILC progenitors. Comparative analysis demonstrated failure to induce various aspects of the ILC gene program in the absence of TOX, implicating this nuclear factor as a key early determinant of ILC lineage specification.
The p38-interacting protein negatively regulates monocyte/macrophage differentiation (HEM5P.239)
Authors: Li Yingqiu , yu xiao , Xu Anlong 
Published: December 2022

Abstract
Monocyte/macrophage differentiation is a central event in the immune response. Here, we investigated the role of p38IP (p38-interacting protein) in monocyte/macrophage differentiation. During monocyte/macrophage differentiation, we found that p38IP is downregulated. Forward knockdown of p38IP by RNA interference induced G1/S arrest, promoting monocytes differentiation into macrophages and the maturation of macrophages as well. Further analysis revealed that p38IP negatively regulates monocyte/macrophage differentiation. Moreover, we found that the downregulation of p38IP is caused by the upregulated miRNA during differentiation. Together, this study reveals the critical role and mechanism for p38IP in regulation of monocyte/macrophage differentiation.
BCAP is a negative regulator of myeloid cell development (HEM5P.225)
Authors: Duggan Jeffrey , Gessay Griffin , Krishnamurty Akshay , Hamerman Jessica 
Published: December 2022

Abstract
B cell adaptor for PI3-kinase (BCAP) is a signaling adaptor protein expressed in hematopoietic cells, including myeloid cells. Here we examined the role of BCAP in myeloid cell homeostasis and development. We found an increased number of monocytes in the bone marrow of BCAP-/- mice versus WT mice, while splenic myeloid populations were similar in number in WT and BCAP-/- mice. Also, mixed chimeras generated with a 1:1 ratio of WT and BCAP-/- bone marrow exhibited a skewing towards BCAP-/- monocytes and neutrophils in the bone marrow, blood and spleen, showing a competitive advantage of BCAP-/- myeloid cells. Thus we hypothesized that BCAP regulates myeloid cell development. WT and BCAP-/- bone marrow had similar numbers of myeloid progenitors, including LSK (Lin-Sca1+cKit+) and CMP (Common Myeloid Progenitor) cells. However, in mixed bone marrow chimeras the progenitor populations were skewed toward BCAP-/- cells, showing that BCAP-/- progenitors out-compete their WT counterparts. In an in vitro myeloid colony forming unit assay, sorted BCAP-/- progenitors produced more cells than WT progenitors, supporting a cell-intrinsic role of BCAP in regulating myeloid differentiation. Also, during infection with Listeria monocytogenes, we observed increased splenic bacterial clearance and numbers of monocytes and neutrophils by day 2 post-infection in BCAP-/- mice compared to WT mice. Together, these data show that BCAP is a negative regulator of myeloid cell development.
Quantitative Studies in Anaphylaxis
Authors: Coulson E. J. , Stevens Henry 
Published: December 2022

Summary
Alum-precipitated ovalbumin had a sensitizing capacity in guinea-pigs of from 4 to nearly 400 times greater than that of ovalbumin in saline solution, depending upon the route of administration of the sensitizing dose.
For ovalbumin in saline solution the sensitizing capacity of the antigen by intraäbdominal administration was about 1½ times greater than by intravenous injection and nearly 2½ times greater that by subcutaneous administration. Only the latter difference was statistically significant.
For alum-precipitated ovalbumin the sensitizing capacity by subcutaneous administration was 2½ times greater than by intraäbdominal administration and 60 times greater than by intravenous injection. Both of these differences were statistically significant.