bluuebunny/crossref_metadata_2025_split huggingface dataset:
Aging subverts immune function by dictating alternative STAT responses to cytokine signaling (P5078)
Authors: Mirza Noweeda , Zorro Manrique Soraya , Cohen Skylar , Dominguez Ana , Cohen Peter , Gendler Sandra 
Published: January 2023

Abstract
Chronic inflammation in the aged closely resembles tumor-induced immune suppression, manifested as a progressive T1→T2 shift and an increased presence of MDSCs. We hypothesize that reversing these age-related phenomena should reduce the elderly’s heightened susceptibility to malignancy. We have now optimized real-time analyses of phosphorylated STATs in murine T cells and MDSCs in order to correlate STAT activation with immune suppression. Both young and old T cells responded to IL-4 stimulation with STAT6 activation and to IFN-γ stimulation with STAT1 activation. Uniquely, however, young T cells also phosphorylated STAT1, STAT3 and STAT5 in response to IL-4, and responded to IL-6 with STAT3 activation, all consistent with a greater susceptibility to T1/T17 differentiation. Regarding MDSCs, both old and young displayed constitutive STAT3 activation which could be enhanced by G-CSF or IL-6 treatment. Both young and old MDSCs activated STAT1 briskly in response to IFN-γ, STAT5 in response to GM-CSF and STAT6 in response to IL-4. However, younger MDSCs, like younger T cells, displayed more diverse responsiveness to IL-4, activating not only STAT6 but also STAT1 and STAT3. While chronic in vivo exposure to T2 cytokines may explain the decreased responsiveness of older MDSCs and T cells to IL-4, paradoxically IL-4 still robustly activated STAT6 in these cells. Such selective responsiveness may be required to preserve the immune suppressive potential of older MDSCs.
Pro-resolving lipid mediators attenuate lung inflammation (P5080)
Authors: Phipps Richard , Thatcher Thomas , Hsiao Hsi-Min , Serhan Charles , Sime Patricia 
Published: January 2023

Abstract
Introduction: Cigarette smoking is the major world-wide cause of COPD and other inflammatory diseases. COPD persists after smoking cessation, suggesting that this may be a failure to resolve inflammation. Resolution of inflammation is an active process regulated by small lipid mediators derived mainly from omega-3 polyunsaturated fatty acids, including the resolvins, and others. We hypothesized that resolvin D1 (RvD1) would have anti-inflammatory and pro-resolving effects in cigarette smoke-induced lung inflammation. Methods: Primary human lung cells (e.g. fibroblasts, etc) were treated with cigarette smoke extract and RvD1 in vitro, and production of pro-inflammatory mediators was determined. Mice were exposed to dilute mainstream cigarette smoke and treated with RvD1 either concurrently with smoke or after smoking cessation. The effect on lung inflammation was assessed. Results: RvD1 suppressed production of pro-inflammatory mediators by human cells. Treatment of mice with RvD1 concurrently with cigarette smoke exposure significantly reduced neutrophilic lung inflammation and production of pro-inflammatory cytokines. RvD1 promoted differentiation of alternatively activated (M2) macrophages. RvD1 also accelerated the resolution of lung inflammation when given after the final smoke exposure. Conclusions: RvD1 has potent pro-resolving effects in cells and mice exposed to cigarette smoke. Resolvins may be a novel therapeutic approach to resolve lung injury and disease.
Novel IL-9-Secreting innate helper cells promote oral antigen-induced anaphylaxis (P6249)
Authors: Lee Jee-Boong , Chen Chun-Yu , Liu Bo , Wang Pin-Yi , Burwinkel Karen , Finkelman Fred D , Hogan Simon P , Wang Yui-Hsi 
Published: January 2023

Abstract
IgE-mediated food allergy is manifested by an overactive T-helper 2 immune response to dietary antigens in the gastrointestinal (GI) tract. Interleukin (IL)-9 promote intestinal mastocytosis which allow the development of oral antigen-induced anaphylaxis. However, little is known about the primary cellular sources of IL-9 or the mechanisms that promote intestinal mastocytosis. Here, we report the identification of multifunctional IL-9-producing innate helper cells (IHC9) that secrete prodigious amounts of TH2 cytokines and mast cell protease-1 (MCPt-1) in response to IL-33 and antigen/IgE complex crosslinking, respectively. Repeated intragastric antigen challenge induces significant accumulations of both intestinal IHC9 and CD4+TH2 cells, which correlate positively with symptoms and susceptibility to oral antigen-induced anaphylaxis. IHC9 development requires T cells and IL-4R, not IL-9R, signaling. The failure of irradiated mice reconstituted with STAT6-deficient bone marrow to generate IHC9 was accompanied by reduced serum MCPt-1, intestinal TH2 inflammation, mastocytosis, and goblet cell hyperplasia and loss of allergic diarrhea. Furthermore, inhibition of food allergy symptoms in mice ablated of IHC9 by anti-FcϵRIα antibody treatment could be reversed by adoptive transfer of IHC9. Thus, IHC9 represent a new type of innate helper cell lineage and function in promoting intestinal TH2 inflammation and mastocytosis that drive IgE-mediated anaphylaxis.
Allergic airway inflammation to cockroach allergen is dependent on thymic stromal lymphopoietin (P6251)
Authors: Sheih Alyssa , Ziegler Steven 
Published: January 2023

Abstract
Environmental allergens are a major culprit in the development and exacerbation of asthma. Particularly, exposure and sensitization to cockroach allergen has been highly associated with asthma severity and morbidity. Currently, there is very little information regarding how exposure to cockroach allergen leads to allergic sensitization and airway inflammation. TSLP (thymic stromal lymphopoietin) is an epithelial cytokine that has been linked to the development of allergic inflammatory diseases. Levels of TSLP are elevated in the lungs of asthmatics and correlates with the severity of the disease. Using a mouse model of allergic airway inflammation to cockroach allergen, we’ve identified an important role for TSLP in the response to cockroach allergen. In this model, intranasal sensitization and challenge to cockroach extract results in a robust TH2 airway inflammatory response along with mucus production. This inflammatory response is significantly reduced in TSLP receptor-deficient mice and depends on the presence of the TSLP receptor on specific immune cell populations. Results from these studies will help expand our knowledge of how exposure to cockroach allergen triggers the development of an allergic TH2 response.
Modulation of dendritic cells by Japanese encephalitis virus: A mechanism of immune evasion (P6344)
Authors: Gupta Nimesh , Hegde Pushpa , Bayry Jagadeesh , Vrati Sudhanshu , Lacroix-Desmazes Sebastien , Kaveri Srini 
Published: January 2023

Abstract
The mechanisms of Japanese encephalitis virus pathogenesis need to be explored to delineate appropriate therapeutic approaches. It is believed that, JEV manipulate the innate and adaptive compartments of the host’s immune system to evade immune response and cross blood-brain barrier to reach the CNS, for pathogenesis and clinical manifestations. The study was designed to explore the functional modulations of dendritic cells and their consequences on CD4+ T lymphocyte polarization after exposure to JEV. Human monocyte-derived DCs were either infected with 1 MOI of virus or mock infected. JEV induced the maturation of DCs with a significant increase in the expression of maturation markers 48 hours post infection, along with inducing the expression of PD-L1 and PD-L2 ligands. Importantly, the JEV-EDIII domain alone was able to reproduce the increased expression of PD-L1 by DCs. JEV-infected DCs induced the generation of Tregs in allogenic mixed lymphocyte reactions. The induction of Tregs by JEV-infected DCs was significantly reduced upon blocking PD-L1 using an antagonist. In addition, JEV-infected DCs significantly altered the proliferation and polarization of T helper cells to ‘Th1/Th2/Th17’ subsets. The results, for the first time, suggest that JEV may evade the host’s immune system by inducing regulatory T cells via PD-L1/PD1 pathway and altering the polarization of T helper cells. The EDIII domain of JEV envelope is implicated in this mechanism of immune evasion.
Memory-like CD8+ T cells that persist throughout MCMV infection have the capacity to self-renew and recapitulate memory inflation (P6312)
Authors: Quinn Michael , Turula Holly , Snyder Christopher 
Published: January 2023

Abstract
Cytomegalovirus (CMV) is a herpesvirses that generates a typically asymptomatic primary infection followed by a life-long latent infection. Episodic viral reactivation events during latency are thought to repeatedly stimulate the immune system and drive the life-long accumulation of functional antigen-specific CD8 T cells, a process termed “memory inflation.” How inflation is homeostatically maintained over time remains unclear. Although the majority of the inflationary pool appears to be terminally differentiated short-lived “effectors” (TEFF) that undergo minimal division during CMV latency, a subpopulation of inflating T cells retains a memory-like phenotype (TEM). We have proposed a model where TEM cells formed early during CMV infection maintain life-long immunity. Utilizing the mouse model, murine (M)CMV, we adoptively transferred TEM cells from chronically-infected mice into naïve mice and rechallenged these cells with MCMV. Interestingly, TEM cells recapitulated inflation by expanding and differentiating asymmetrically to produce TEFF and TEM progeny. This process appears influenceable by factors known to modulate terminal differentiation (i.e. antigen load and inflammation). Importantly, the TEM progeny retained their “memory capacity” and could reproduce memory inflation even after repeated rechallenges.
Invariant natural killer T cells recognize and restrict the transformation of B cells by Epstein-Barr virus (P4363)
Authors: Chung Brian , Tsai Kevin , Gibson William , van den Elzen Peter , Priatel John , Tan Rusung 
Published: January 2023

Abstract
Individuals with X-linked lymphoproliferative disease (XLP) lack invariant natural killer T (iNKT) cells and are uniquely susceptible to Epstein-Barr virus (EBV). To determine whether iNKT cells recognize or regulate EBV replication, human B cells were infected with GFP+ EBV 95.8 in the presence or absence of iNKT cells. We observed that iNKT cells significantly reduced early EBV titres and GFP+ B cell expansion but that within 72 hours of infection, B cells transformed into lymphoblastoid cell lines (LCL) by EBV had downregulated the iNKT antigen receptor CD1d, completely abrogating iNKT recognition of the lipid agonist α-galactosylceramide (αGalCer). To determine whether ectopic CD1d expression could restore iNKT recognition, LCL were treated with a synthetic retinoic acid receptor-α agonist AM580 known to modulate CD1d expression by regulating the nuclear protein lymphoid enhancer-binding factor-1 (LEF-1). AM580 treatment significantly reduced LEF-1 at the CD1d promoter, induced surface CD1d expression and enabled iNKT recognition of LCL loaded with αGalCer. Unexpectedly, AM580-treated LCL also activated iNKT function even in the absence of αGalCer suggesting that EBV infection and transformation triggers endogenous iNKT antigen expression. These observations indicate that iNKT cells may be important for the early recognition and control of EBV-infected B cells prior to CD1d downregulation and the loss of CD1d may allow EBV to circumvent iNKT-mediated immunosurveillance.
A CpG triggered Syk-Calcium-CaMKII signal directs the secretion of TNFα by innate immune cells (P6297)
Authors: Behrens Edward , Liu Xiaohong , Freedman Bruce , Rao Sheila 
Published: January 2023

Abstract
The kinase Syk is reported to regulate cytokine secretion in multiple and conflicted manners. We therefore examined Syk in regulating CpG DNA stimulated TNFα secretion. Secreted cytokines were measured by ELISA and surface cytokines by flow cytometry. Gene deletion was achieved with both Cre-Lox and shRNA methods. Deletion of Syk decreases CpG induced TNFα secretion while TNFα mRNA and intracellular protein levels remain normal. In contrast, IL-6 secretion is unaffected. CpG induced plasma membrane levels of TNFα are decreased in Syk deficiency, suggesting that Syk is important for TNFα intracellular trafficking. A downstream effector of Syk is calcium (Ca) flux mediated by PLCγ activation. Supporting a role for Ca in Syk mediated TNFα release, we show: 1) CpG stimulation triggers calcium flux only in Syk-sufficient cells, 2) silencing PLCγ2 phenocopies the TNFα defect of Syk-deficient cells, 3) forced Ca mobilization using ionomycin rescues the TNFα secretion defect in Syk-deficient cells. CpG-triggered phosphorylation of the calcium dependent kinase CaMKII is defective in Syk-deficient cells. CpG-induced TNFα secretion is defective in CaMKII-deficient cells, and is not rescued by ionomycin, suggesting that CaMKII is downstream in the pathway initiated by Syk signaling. These data suggest that CpG provides active signals for TNFα transcription and translation as well as intracellular trafficking, via a Syk-Ca-CaMKII axis, allowing for the fine-tuning of TNFα secretion.
The splicing factor SF2/ASF is a novel regulator of IL-2 transcription and rescues IL-2 production in T lymphocytes from patients with SLE (P6288)
Authors: Moulton Vaishali , Grammatikos Alexandros , Fitzgerald Lisa , Tsokos George 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease, which afflicts mainly women in the childbearing years, and causes painful arthritis, skin disease and complications in the kidneys and brain. Abnormal T lymphocytes in SLE not only regulate autoantibody-producing B cells, but also are responsible for target organ infiltration. T cells from patients with SLE express decreased levels of the T cell receptor (TCR) - associated CD3 zeta (ζ) chain, a feature directly linked to their poor interleukin (IL)-2 production. We recently showed that the splicing regulator splicing factor 2 / alternative splicing factor (SF2/ASF) enhances the expression of CD3ζ chain by limiting the production of an unstable mRNA splice variant. Here we show that SF2/ASF expression levels are decreased in T cells from SLE patients, more so in patients with higher SLE disease activity index (SLEDAI). Importantly, we show that overexpression of SF2/ASF in SLE T cells restores IL-2 production. In parallel, silencing SF2/ASF expression in normal T cells reduces their IL-2 secretion. Finally we show that SF2/ASF induces transcriptional activity of the IL-2 promoter and that SF2/ASF is recruited to the IL-2 promoter indicating its role in IL-2 transcription. These results identify SF2/ASF as a novel regulator of IL-2 expression in human T cells and a potential molecular mechanism underlying the T cell defect in SLE.
Two functional ligand binding sites on human CD30 separately located from the epitope of agonistic monoclonal antibodies (P6339)
Authors: Ise Tomoko , Abe Yasuhiro , Nagata Satoshi 
Published: January 2023

Abstract
Human CD30 is a member of the tumor necrosis factor (TNF) receptor superfamily (TNFRSF8). CD30 is considered to exist as homotrimers on the cell surface without binding to the ligand. Several anti-CD30 agonistic antibodies have been isolated. One of them is currently clinically used as an antibody drug conjugate for therapy of CD30-positive lymphomas. Despite progress in clinical application, the binding mode required for the agonistic activity of anti-CD30 antibodies has not been fully understood. In particular, the presence of two duplicated sequences including the TNF receptor domain in human CD30 complicates the analysis of binding-function relationships. In this study, we produced a series of recombinant conformation-carrying CD30 mutants whose TNF receptor domains and epitopes are replaced with corresponding sequences of CD40, based on the structures. We tested their ability to transduce signals for NFkB activation in reporter cells upon binding to CD30-ligand or agonistic antibodies. We found that both of the predicted ligand binding sites activate NFkB pathways by the binding of CD30-ligand-Fc fusion protein or CD30-ligand expressed on 293T cells. In addition, a group of agonistic antibodies appeared to bind an N-terminus site of the non-ligand binding site mutants and activated NFkB. Our results suggest that agonistic antibodies induce the active complex formation of membrane CD30 for potent signaling through a different binding mode than the natural ligand.
C-C chemokine receptor regulates the antibody response to DNP-KLH (P6351)
Authors: Vines Charlotte , Welch T. , Fang Li , Kaiser Brian , Islam Shamima , Bill Charles , Sun Yuning , McBurney Tiffany 
Published: January 2023

Abstract
Functional loss of C-C chemokine receptor 7 (CCR7) and its ligands, CCL19 and CCL21, has been linked to development of autoimmune diseases. Therefore, we hypothesized that CCR7 antagonists could be used to promote an immune response that could be measured by tittering antibodies produced in animals injected with test antigens. To test this hypothesis we injected C57BL/6 mice on day 0 and boosted on day 20 with dinitrophenol (DNP)-Keyhole Limpet Hemocyanin (KLH) with a CCL19 antagonist, 8-83, or with the agonist solvent as a control. Similar to the CCR7-/- mouse, which has significantly elevated levels of IgG1, IgG2a, IgG2b and IgG3 by day 30, the animals treated with 8-83 generated significantly higher levels of IgG1 and IgG2a, than animals injected with the DNP-KLH/solvent alone. Previous studies by our laboratory have shown that CCL19 signals through ERK5. To determine if loss of signaling through ERK induced a similar increase in the levels of IgGs, we injected ERK5flox/flox control mice or ERK5flox/floxLck-Cre mice with DNP-KLH. Similar to animals treated with 8-83, the ERK5flox/flox Lck-Cre mice produced significantly higher levels of IgG1, IgG2a and IgG3 than control mice. From these results we conclude that CCR7/CCL19 signals through ERK5 to control the levels of antibodies made. Recent studies demonstrating a role for CCR7 in controlling the localization of regulatory T cells may provide insight into immune cells that are regulate observed increase in antibody titers.
IL-10-producing B cells limit CNS inflammation and infarct volume after experimental stroke in mice (P1090)
Authors: Offner Halina , Chen Yingxin , Murphy Stephanie , Bodhankar Sheetal 
Published: January 2023

Abstract
Clinical stroke induces inflammatory processes leading to cerebral injury. IL-10 expression is elevated during major CNS diseases and limits inflammation in the brain. Recent evidence demonstrated that absence of B cells led to larger infarct volumes and increased numbers of activated T cells, monocytes and microglial cells in the brain, thus implicating a regulatory role of B cell subpopulations in limiting CNS damage from stroke. The aim of this study was to determine whether the IL-10-producing regulatory B cell subset can limit CNS inflammation and reduce infarct volume following ischemic stroke in B-cell deficient (μMT-/-) mice. Five million IL-10-producing B cells were obtained from IL-10-GFP reporter mice and transferred i.v. to μMT-/- mice. After 24 h from this transfer, recipients were subjected to 60 min of middle cerebral artery occlusion (MCAO) followed by 48 h of reperfusion. Compared to vehicle-treated controls, the IL-10+ B-cell-replenished μMT-/- mice had reduced infarct volume and fewer infiltrating activated T cells and monocytes in the affected brain hemisphere. These effects in CNS were accompanied by significant increases in regulatory T cells and expression of the co-inhibitory receptor, PD-1, in the periphery. These novel observations demonstrate both immunoregulatory and protective functions of IL-10-secreting B cells in MCAO that potentially could impart significant benefit for stroke patients in the clinic.
Generation of mature activated regulatory myeloid cells: Differential effects of retinoic acid on myelopoiesis versus dendropoiesis (P1064)
Authors: VanGundy Zachary , Baker Julie , Stranger Heather , Papenfuss Tracey 
Published: January 2023

Abstract
Myeloid cells are increasingly recognized for their potent regulatory abilities. However, factors influencing regulatory myeloid cell (regMC) differentiation remain poorly understood. Retinoic acid (RA) is a steroid hormone important in regulating mucosal (gut) immunity. RA also promotes myeloid differentiation. We hypothesize that RA during differentiation will promote regMC formation (both regulatory DCs and macrophages). Using in vitro differentiation of myeloid cells with GM-CSF with or without RA, we found that day 6-7 RA BM-MCs were more mature/activated (i.e. increased expression of CD80, CD86, and MHCII) than control BM-MCs. Importantly, RA BM-MCs also had increased expression of the inhibitory marker PD-L1. Functionally, these RA BM-MCs expressed increased intracellular IL-10 and induced an increased percentage of T regulatory cells in vitro compared to controls. Notably, RA BM-MCs suppressed the proliferation of responder immune cells even after inflammatory (LPS) challenge. Surprisingly, CD11c+ (DCs) within the RA BM-MC population were not the suppressive population. Rather, the suppressive cells were CD11c-CD11b+Ly6Cint/low and likely represent a regulatory monocyte or other myeloid precursor population. Additional studies characterizing these cells are ongoing. These results suggest that RA has differential effects on myelopoiesis versus dendropoeisis which produce non-DC regMCs.
Cell hybridization technology produces monoclonal antibodies against MurA protein of Listeria monocytogenes (P3132)
Authors: Wen Yiming , Li YuChen , Li Zhiqing , Tong Jiyu , Yang Hongyu , Xiang Junjian 
Published: January 2023

Abstract
The research aims to develop and characterize monoclonal antibodies(MAbs) with high specificity and affinity for surface antigens of all major serotypes of Listeria monocytogenes(L. monocytogenes). Hybridomas were produced using extraction of L. monocytogenes surface protein as the immunogen. Detected by enzyme-linked-immunosorbent assay (ELISA), MAb1B10 was capable of binding to cell-surface antigens of live L. monocytogenes. MAb1B10, 2C10 and 1G3 reacted with both formalin-inactivated and heat-killed L. monocytogenes without any cross reactions with other Listeria spp.; MAb1C10 and 3G8 highly reacted with L. monocytogenes, L. innocua and L. welshimeri, but seldom reacted with non-Listeria spp.. Immunoprecipitation-mass spectrometry (IP-MS) and western blot were applied to convince that the reactive epitope was on the MurA protein of 72kDa. Recombinant MurA protein produced in Escherichia coli was confirmed to be the antigen which was recognized by MAb1B10, 2C10 and 1G3. To sum up, MAbs reacting with live L. monocytogenes, formalin-inactivated and heat-killed L. monocytogenes were produced. The results indicated that MurA could be exploited as a marker for specifically identifying pathogenic L. monocytogenes in suspect food samples.