bluuebunny/crossref_metadata_2025_split huggingface dataset:
ILC3 detected in intestinal mucosal tissues of mice with chronic alcohol consumption (IRC4P.608)
Authors: Ito Ichiaki , Nishiguchi Tomoki , Asai Akira , Suzuki Sumihiro , Kobayashi Makiko , Suzuki Fujio 
Published: January 2023

Abstract
Bacterial translocation from microbiota causes serious sepsis in mice with chronic alcohol consumption (CAC-mice). Since innate lymphoid cells (ILCs) are cells to be involved in the prosperity and decoy of host antibacterial innate immunities, in this study, we tried to isolate ILCs from intestinal mucosal tissues of CAC-mice with various durations of alcohol consumption. CAC-mice were prepared by an oral gavage of 0.6 ml/day of 20% EtOH for 4 to 14 weeks. Lineage negative (lin-) cells were isolated from the intestinal mucosal tissue of CAC-mice. These cells were stimulated with PMA (50 ng/ml) and ionomycin (500 ng/ml) for 6 hours. Obtained cells were analyzed for ILC2 and ILC3 by flow cytometry, and culture fluids of lin- cells were assayed for IL-22 and IL-17A by ELISA. In the results, the percentage of CD45+CD90+RORγt+ cells (ILC3) in the lin- cell population was gradually increased in mice with alcohol consumption for 10-12 weeks (7.84% in average), as compared to be percentage of ILC3 in normal mice (0.67%). On the other hand, CD127+ICOS+Sca-1+ST2+ cells (ILC2) were minimally isolated from the lin- cell population of 4-12 week CAC-mice. Lin- cells from 12 week CAC-mice produced 400 pg/ml of IL-22 and 1.7 ng/ml of IL-17A. These results indicate that 10-12 week CAC-mice are carriers of ILC3 which play a role in the increased susceptibility of CAC-mice to enterococcal translocation and subsequent sepsis.
Differential gene expression following stimulation of TLRs 2, 3, 7 and 8 in rag1-/- mutant zebrafish (IRC4P.613)
Authors: Muire Preeti 
Published: January 2023

Abstract
Natural Killer (NK) cells have shown to mediate adaptive immune responses to chemical haptens and cytomegalovirus in T and B cell deficient mice. T and B cell deficient zebrafish mount specific protection to intracellular bacteria. We hypothesize that a sub-population of zebrafish NK cells mediates this protection, and these cells have a mechanism for enhanced discrimination of a bacterial target. However, zebrafish NK cells have not been characterized, and their response to TLR agonists has not been investigated. We utilized rag1-/- mutant zebrafish as an in vivo fish model to determine if TLR agonists (TLR2: βglucan, TLR3: Poly I:C and TLR 7/8: R-848) stimulate NK cells in liver, kidney marrow and spleen tissues. We used IFNγ expression as an indicator of NK cell activation, TNFα expression as an indicator of macrophage activation, and MxA expression as an indicator of IFNa and IFNβ production. With RT-qPCR we quantified transcript levels at 1h, 6h, 12h and 24h after injecting rag1-/- mutant zebrafish with either βglucan, POLY I:C or R-848. Our results show significant increases of IFNγ, MxA and TNFα transcripts in the three tissues with βglucan, POLY I:C or R-848 compared to PBS injected control fish. Liver had the highest expression of IFNγ in response to R-848 and βglucan. The data presented are the initial findings in an investigation to help define mechanisms involved in NK cell mediated protective immunity in rag1-/- mutant zebrafish.
Cholinergic modulation of inflammatory diseases: role in the amelioration of type 1 diabetes (IRC4P.452)
Authors: Fernandez-Cabezudo Maria , George Junu , Bashir Ghada , Al-Ramadi Basel 
Published: January 2023

Abstract
The cholinergic anti-inflammatory pathway connects the central nervous system with the immune system via the vagus nerve and controls the release of pro-inflammatory cytokines. We have previously shown that inhibition of acetylcholinesterase (AChE), thereby leading to an accumulation of acetylcholine (ACh), modulates the inflammatory response. Type 1 diabetes (T1D) results from a deficiency in the production of insulin as a consequence of an immune-mediated damage of pancreatic β cells. Here, we investigated the potential of regulating the development of T1D through the inhibition of AChE. Modulation of AChE activity was achieved by administration of paraoxon, a potent inhibitor of the enzyme. To induce hyperglycemia, we used a well-characterized regimen of multiple low doses of streptozotocin (MLD-STZ). Our results showed that inhibition of AChE prevented the development of MLD-STZ-induced hyperglycemia (blood glucose levels &gt;200 mg/dl) in C57BL/6 mice. At day 18 post-STZ administration, immunohistological analysis of pancreatic tissue in paraoxon pre-treated mice revealed healthy islets with normal levels of insulin. In contrast, control mice routinely exhibited glucose levels of 300-400 mg/dl and severe loss of islet morphology by the same time point. qRT-PCR analysis demonstrated that AChE inhibition prevents the loss of insulin production induced by MLD-STZ administration. Our results provide evidence for cholinergic control of the development of T1D in mice.
IL-25 promotes the function of CD4+CD25+ T regulatory cells and prolongs skin-graft survival in murine models (IRC5P.620)
Authors: Zhou Xiaohui , Tang Jiayou , Liu Jie , Meng Qingshu , Liu Zhongmin , Fan Huimin 
Published: January 2023

Abstract
Interleukin (IL)-25, also known as Interleukin (IL)-17E, belonging to the IL-17 family. Unlike the other members, IL-25 has long been deemed as a pro-inflammatory cytokine favoring Th2--type immune response. Here, we show that expression of IL-25 in skin and subcutaneous tissues were constantly, while increased during allograft rejection. At same time, deficiency of IL-25 aggravates skin allograft rejection in murine models. Further experiments showed deficiency of IL-25 down-regulated the expression of Foxp3 among CD4+ cells and aggravated skin allograft rejections in murine models. While, exogenous IL-25 restored the Foxp3 expression and the function of Tregs in vitro. Adoptively transfer of IL-25 deficient Tregs failed to protect the allograft from rejection, compared to the WT Tregs. Moreover, IL-25 promoted phosphorylation of NFAT2, up-regulated the Foxp3 expression and contributed to the function of Tregs. Our findings indicate that IL-25 can promote the function of Tregs via impeding the interaction of NFAT2 and Foxp3, and prolong skin-graft survival.
TSG6 as a central mediator of macrophage M2 polarization (IRC5P.626)
Authors: Mittal Manish , Toya Sophie , Malik Asrar 
Published: January 2023

Abstract
TNF-α stimulated gene-6 (TSG6) is an anti-inflammatory protein, secreted by activated monocytes and endothelial cells. TSG6 binds to CD44 receptor expressed on multiple cell types including monocytes/macrophages. We tested the concept that TSG6 via binding to CD44 induces a phenotypic shift in macrophages from pro-inflammatory M1 to anti-inflammatory M2 type that protects the mice against lung inflammation and injury induced by endotoxin LPS. We show that TSG6 suppressed the induction of M1 markers (IL-6, IL-1β, CXCL1, iNOS) and activated expression of M2 marker genes (such as IL-10, Mannose receptor (CD206) and Chitinase-3-like-3). The shift was abrogated in macrophages isolated from CD44-/- mice. TSG6 also suppressed the activation of NF-ΚB signaling by inhibiting interaction of MyD88 with TLR4. In addition, TSG6 induced activation of anti-inflammatory transcription factor STAT6 in LPS exposed macrophages. Intra-tracheal instillation of TSG6 protected the mice against LPS induced mortality. Moreover, mice treated with TSG6 showed significantly suppressed lung vascular injury and neutrophil sequestration on challenge with LPS. That TSG6 binding to CD44 activates the macrophage anti-inflammatory program suggests that TSG6 is a specialized regulator of inflammation and potential therapeutic target.
C1q-based tolerance mechanism in HMGB1-mediated activation of human monocytes (IRM5P.649)
Authors: Son Myoungsun , Porat Amit , Tracey Kevin , Diamond Betty 
Published: January 2023

Abstract
C1q, a component of the classical complement pathway, is required for clearance of apoptotic cells. C1q also regulates activation of innate immune cells through engagement of C1q receptors present on the immune cell surface. C1q deficiency is highly associated with the development of systemic lupus erythematosus (SLE). High Mobility Group Box 1 (HMGB1), a pro-inflammatory mediator, is secreted by inflammatory cells and passively released from necrotic cells. HMGB1 interacts with TLR4 and receptor for advanced glycation end products (RAGE), to augment the production of type 1 interferon and proinflammatory cytokines. Here we show that C1q inhibits HMGB1-mediated monocyte activation through cross-linking RAGE and Leukocyte-associated Ig-like receptor-1 (LAIR-1), which is an inhibitory receptor for collagen and C1q. This crosslinking causes LAIR-1 to enter lipid rafts following HMGB1 and C1q stimulation. C1q suppress HMGB-1 mediated induction of type I-interferon and pro-inflammatory cytokines including IL-12 and TNF-α. Moreover, exposure of monocytes to HMGB1 and C1q together leads to upregulation of a number of anti-inflammatory molecules that might function to induce phagocytosis and to maintain monocyte quiescence. Our results suggest that C1q regulates the activation of innate immunity by HMGB1 and maintains immune homeostasis. These data are consistent with the established requirement for C1q in the prevention of autoimmunity.
Bcl6 mediates the regulation of the innate immune response against RNA virus (IRM5P.650)
Authors: Shi Liyun , Kang Yanhua , Zhuang Ningtong , Zhang Hang 
Published: January 2023

Abstract
Bcl6, a POZ transcription factor, plays an essential role in the tumorgenensis, inflammation and development, but its role in the antiviral response has never been established. Here, we demonstrate that Bcl6 unexpectedly functions to inhibit RIG-I-mediated antiviral signaling and impedes the clearance of viral infection. More importantly, we identify that miR-127 as a upstream regulator of Bcl6, and the subsequent co-repressive complex in the initiation of RIG-I-driven response. This finding extends our knowledge about mitochondrial antiviral mechanisms and proposes a novel corepressor network required for the coordination of the innate immune response.
p53 regulates expression of the early response gene tumor necrosis factor-α-induced protein 8 in macrophage (IRM6P.660)
Authors: Lowe Julie , Menendez Daniel , Nguyen Thuy-Ai , Grim Sara , Resnick Michael , Fessler Michael 
Published: January 2023

Abstract
The cell response to pathogens involves complex signaling cascades that affect expression of thousands of genes. Characterization of the regulation and function of genes involved in the stress response is critical since failure of this response can foster the development of chronic disorders such as autoimmune diseases and cancer. A growing body of evidence shows a role for the transcription factor p53 in the immune response. To better characterize the role of p53 in the innate immune response, we performed a microarray in primary human macrophages and identified the gene Tumor Necrosis Factor-Alpha Induced Protein-8 (TNFAIP8) as novel p53 target. ChIP-seq analysis shows strong binding of p53 to an intragenic region within TNFAIP8, which contains a functional p53 response element as confirmed with reporter assays. This region is an enhancer, since ChIP assays show enhancer-like histone modifications and 3C assays indicate chromatin looping with the promoter region. Very little is known about TNFAIP8 function; however, Tnfaip8 was previously reported to confer resistance to Staphylococcus aureus, suggesting a role in host defense. Studies are currently underway to characterize expression regulation and functions of TNFAIP8 in macrophages in response to TLR ligands. These results provide a better understanding of the role of p53 in the innate immune response and the regulation and function of the understudied TNFAIP8 gene.
A dichotomy in cellular immune responses to experimental blood stage malaria infection in humans revealed by systems immunology (IRM6P.661)
Authors: Burel Julie , Apte Simon , Mccarthy James , Doolan Denise 
Published: January 2023

Abstract
Despite intense research for many decades, the mechanisms of protective immunity against malaria are poorly understood. Recent technological and conceptual advances in the field of immunology, in particular Systems Immunology, offer the potential to provide new insights into the key mechanisms involved in host immunity. This study aimed to dissect at the molecular level the cellular immune responses to malaria by combining miRNA and mRNA profiling, multiparameter flow cytometry with a unique P. falciparum blood stage experimental infection model. Unexpectedly, naive subjects segregated into two distinct groups based on their molecular immune profile upon infection, classified as High and Low responders. High responders presented increased expression of circulating miRNAs involved in immune response, increased lymphocyte activation, increased P. falciparum specific antibody responses and reduced blood stage parasitemia over the course infection; no significant changes were observed in Low responders. This study is the first to demonstrate inter-individual variability in immune responses to experimental malaria infection. The molecular profile of circulating lymphocytes prior to infection in High and Low responders is currently being investigated, to shed light on the molecular mechanisms driving this dichotomy in immune responses. This may predict immune response in naive individuals and therefore would have tremendous implications for vaccine development and evaluation.
The RNA-binding protein, polypyrimidine tract-binding protein, affects multiple events in CD4 T cell activation through distinct processes (IRM7P.717)
Authors: Covey Lori , La Porta James , Matus Nicodemos Rodrigo 
Published: January 2023

Abstract
We have previously shown that the RNA binding protein, polypyrimidine tract-binding protein (PTB) plays a crucial role in regulating the expression of CD40L in activated CD4 T cells. Regulation occurs through stabilizing the message at late times of activation as well as changing the distribution of CD40L mRNA within distinct cellular compartments. Further experiments were designed to assess how PTB affects global CD4 T cell activation by using shRNA introduced into primary human CD4 T cells. Specifically, we found that PTB is critical for the proliferation and viability of activated T cells. The propensity to undergo cell death was evident in both infected and uninfected CD4 T cells within the same population suggesting that either soluble and/or cognate interactions were being affected by decreased PTB. Accordingly, PTB was found to be required for optimal expression of CD25, CD69, TNF-α and IL-2 but not CD38 or IFN-γ. When the decay rates were assessed in anti-CD3/anti-CD28-stimulated CD4 T cells, the pattern of IL-2, TNF-α and CD69 mRNA decay did not change over a time course of activation whereas CD25 showed a pattern of mRNA decay that was highly similar to CD40L. Evaluation of signaling pathways using phosphoflow analysis revealed PTB-specific changes in the NF-κB and MAPK-ERK pathways. Together, these findings support a role for PTB in the control of CD4 T cell activation through processes that are both dependent on and independent of PTB-regulated mRNA stability.
Nedd4 family interacting proteins activate ubiquitylation pathways to limit ERK signaling following TCR engagement (IRM7P.702)
Authors: O'Leary Claire , Riling Chris , Deng Guoping , Spruce Lynn , Seeholzer Steve , Oliver Paula 
Published: January 2023

Abstract
Ubiquitylation tunes signaling pathways in stimulated T cells to regulate activation and function. We have found that, in vitro, Nedd4-family interacting protein 1 (Ndfip1) and Ndfip2 activate Nedd4-family catalytic E3 ligases with known roles in T cells. Ndfip1 negatively regulates T cell activation and Th2 polarization, but the in vivo role of Ndfip2 is unknown. We generated Ndfip2-/- mice and found that Ndfip2 is not a prominent negative regulator of T cell activation or Th2 polarization. However, loss of Ndfip2 exacerbates the inflammatory Ndfip1-/- phenotype, suggesting that Ndfip2 dampens inflammation. We generated mixed fetal liver chimeras and found that Ndfip1/Ndfip2 deficiency leads to T cell intrinsic hyperactivity. To identify signaling pathways affected by loss of Ndfips, we used semiquantitative proteomics to compare stimulated control and knockout CD4+ T cells. Pathway analysis revealed increased MEK1 activity in knockout cells, which was confirmed by observing increased ERK phosphorylation in stimulated Ndfip1/Ndfip2 deficient CD4+ T cells. To identify aberrant ubqiuitylation of proteins upstream of ERK, we have taken a quantitative proteomic approach, using Tandem Ubiquitin Binding Entities (TUBEs) to enrich for polyubiquitylated proteins in isotope labeled control and Ndfip deficient CD4+ T cells. This approach has yielded several potential targets of Ndfip-dependent ubiquitylation in CD4+ T cells that are currently under investigation.
CD5 inhibits the activity of dendritic cells to activate T cells and induce immune responses (IRM7P.703)
Authors: Xu Hui , Li Hui , He Donggou , Xu Shin , Raman Chander 
Published: January 2023

Abstract
It is known that CD5 mediates inhibitory signals in T cells and plays important roles in immune tolerance. Little is known about whether CD5 has a role in dendritic cell (DC) induced T cell activation and immune responses. Here we show that CD5 was expressed by CD11c+ DC in the thymus, lymph node, spleen and skin of mice. A deficiency in CD5 gene (CD5-/-) did not have a significant effect on the development of DC. However, it enhanced the activity of DC to activate CD4 and CD8 T cells in vitro and in vivo. CD5-/- DC were more potent than wild type DC in the induction of anti-tumor immunity and contact hypersensitivity response in animals. CD5-/- DC induced higher levels of IL-2 and IFN-gamma production by T cells than wild type DC. Restoration of CD5 expression on CD5-/- DC inhibited their activity to activate T cells and induce immune responses in mice. Further analysis indicated that the deficiency in CD5 increased the production of IL-12 and IL-23 by DC and that the restoration of CD5 expression inhibited the cytokine production by CD5-/- DC. Collectively, the study has demonstrated that CD5 inhibits the production of inflammatory cytokines by DC and has an inhibitory effect on their activity to activate T cells and induce immune responses. It provides novel insights into mechanisms for DC induced T cell tolerance and immune suppression.
CD28 induces mitochondrial respiration through Slp-76 in long-lived plasma cells for reactive oxygen species dependent survival. (LYM2P.737)
Authors: Utley Adam , Carlson Louise , Rozanski Cheryl , Lee Kelvin 
Published: January 2023

Abstract
Sustained humoral immunity is dependent upon bone marrow (BM) resident long-lived plasma cells (LLPC). We have shown that CD28 signaling in LLPCs is necessary for their survival and antibody production; however, the mechanistic basis for this is unclear. In T cells, CD28 induces glycolysis at the expense of mitochondrial respiration. Surprisingly, CD28 activation drives mitochondrial respiration in LLPCs but not short-lived plasma cells (SLPC) through PLCγ1. Paradoxically, respiration dependent production of reactive oxygen species is required for CD28 mediated survival. Mechanistically, CD28 provides a pro-survival signal in LLPCs through Slp-76, an adaptor protein known to be upstream of PLCγ1 in T cells. CD28 itself drives Slp-76 transcription and subsequent protein expression. Additionally, at the mRNA level Slp-76 is highly expressed in human BM PCs (LLPCs) when compared to splenic PCs (SLPCs). Furthermore, CD28 upregulates the APRIL/BAFF receptor BCMA in LLPCs but not SLPCs. Due to the high levels of Slp-76 seen in LLPCs, but not in SLPCs, we propose a model wherein CD28 induces a longevity program through Slp-76 during plasma cell differentiation that facilitates plasma cell responsiveness to extrinsic survival signals in the bone marrow niche, allowing for a plasma cell to become long-lived. By doing so, CD28 may regulate LLPC induction, function and survival. CD28 therefore represents a novel signaling axis to exploit for the augmentation of vaccine design.
Role of IL-4Rα on myeloid dendritic cells in pathogenicity of respiratory syncytial virus infection and asthma development (MUC1P.909)
Authors: Shrestha Bishwas , You Dahui , Saravia Jordy , Siefker David , Cormier Stephania 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract infections in infants and severe disease is associated with asthma development in later life. We have shown that neonatal mice respond to primary RSV infection with T helper type 2 (Th2) biased immune responses which are enhanced following reinfection. Our lab has demonstrated elevated levels of IL-4Rα on neonatal mDCs compared to adults. Therefore, we investigated the role of IL-4Rα on mDCs in the RSV induced Th2 biased response in neonates. CD4+ T cells from adult mice were co-cultured with RSV infected CD11c+ cells (DCs) from neonatal wild type or IL-4Rα-/- mice and resulting CD4+ T cell sub phenotypes were analyzed. DCs from IL-4Rα-/- mice induced more Th1 and less Th2 cells compared to DCs from wild type mice. Thus, we hypothesized that IL-4Rα on neonatal mDCs is responsible for the pathogenicity in RSV infection. To test this hypothesis, we used transgenic mice in which IL-4Rα is specifically depleted on CD11c+ cells. These mice were infected with RSV as neonates and reinfected four weeks later. We then analyzed the pulmonary T cell profile, cytokines, and pathophysiological responses. Mice in which IL-4Rα was deleted on mDCs showed significant reductions in their pathological Th2 responses. Together, these data demonstrate that the asthmatic Th2 biased immune phenotype induced during neonatal RSV infection is mediated in part by IL-4Rα expression and signaling on mDCs.
Transfer factor: forgotten, but not gone (MUC2P.934)
Authors: Myles Ian , Datta Sandip 
Published: January 2023

Abstract
It has been over 70 years since the discovery of cellular immunity. Further investigation revealed that the even if immune cells are lysed and the molecular size limited to 8kDa, such injections retain some of the immunity-transferring properties. The term “transfer factor” (TF) was coined to denote the undiscovered substance responsibly for this phenomenon. Yet despite early interest, TF was all but abandoned in the 1990’s; in part due to unsubstantiated and fraudulent claims as well as the discovery of HIV, which suspended consideration of deriving therapy from blood. However, while TF was shelved as poorly elucidated, no clear explanation was provided for the observation of a small molecule harboring antigen-specific properties and therapeutic potential. Reopening this chapter, we discovered transfer factor is produced by memory T-cells and acts directly upon dendritic cells in an antigen-specific manner; ultimately activating mucosal immunity via Tc17 pathways. Antigen-targeted TF enhanced inflammatory response in models against Candida as well as S. aureus. Mass spec data suggests TF is hybrid compound, predicted to containing elements of peptides and nitrogenous bases. Finally, we provide ex vivo human data to demonstrate the therapeutic potential of TF and even attempt to address some of the past’s more fanciful claim. This report represents transfer factors’ resurrection and provides a modern scientific investigation into this once forgotten area of immunology.