bluuebunny/crossref_metadata_2025_split huggingface dataset:
Suppression of IL-12 production by TTP through blocking NF-kappaB nuclear translocation (P3008)
Authors: Liu Jianguo , Gu Ling , Ning Huan , Hou Rong 
Published: January 2023

Abstract
Tristetraprolin (TTP) plays a significant role in regulating the expression of AREs containing mRNAs. TTP-/- Mice develop a systemic autoimmune inflammatory syndrome characterized by cachexia, conjunctivitis and dermatitis. IL-12 plays a crucial role in immune defense against infectious and malignant diseases. In the present study, we found increased production of IL-12 during endotoxic shock and enhanced Th1 cells in TTP knockout mice. The levels of IL-12 p70 and p40 protein as well as p40 and p35 mRNA were also increased in activated macrophages deficient of TTP. In line with these findings, overexpression of TTP suppressed IL-12 p35 and p40 expression at the mRNA and promoter level, while surprisingly had no effects on their mRNA stability. Our data showed that the inhibitory effects of TTP on p35 gene transcription were completely rescued by overexpression of NF-kappaB p65 and c-Rel but not the p50 in activated macrophages. Our data further indicated that TTP acquired its inhibition on IL-12 expression through blocking nuclear translocation of NF-kappaB p65 and c-Rel while enhancing p50 upon stimulation. In summary, our study reveals a novel pathway through which TTP suppresses IL-12 production in macrophages, resulting in suppression of Th1 cell differentiation. This study may provide us with therapeutic targets for treatment of inflammatory and autoimmune disorders
Fli-1 drives transcription from the MCP-1 gene promoter, which may provide a novel mechanism for cytokine and chemokine activation. (P6236)
Authors: Lennard Richard Mara , Nowling Tamara , Zhang Xian 
Published: January 2023

Abstract
Regulation of proinflammatory cytokines and chemokines is a primary role of the innate immune response. MCP-1 is a chemokine that recruits immune cells to sites of inflammation. Expression of MCP-1 is reduced in primary kidney endothelial cells from mice with a heterozygous knockout of the Fli-1 transcription factor. Fli-1 is a member of the Ets family of transcription factors, which are evolutionarily conserved across several organisms including Drosophilla, Xenopus, mouse and human. Ets family members bind DNA through a consensus sequence GGAA/T, or Ets binding site (EBS). We have demonstrated that Fli-1 binds to three EBSs within the endogenous MCP-1 promoter by ChIP assay. Transient transfection assays of the murine MCP-1 promoter indicate that the Fli-1 gene actively drives transcription from the MCP-1 gene promoter in a dose-dependent manner. The Ets-1 transcription factor was also tested, but failed to drive transcription. Studies to determine the relationship between Fli-1 and Ets-1 in the transcriptional control of MCP-1 will be presented. Deletion studies to determine the most important regions of the MCP-1 promoter for Fli-1 gene regulation are being investigated. Together, these results demonstrate a novel pathway, separate from NFκB, for the transcriptional activation of MCP-1. Evidence suggests that Fli-1 may interact with the promoters of other chemokines and cytokines, providing a novel mechanism for activation of the inflammatory response.
Pharmacokinetics and immunological effects of human IL-15/IL-15Rα heterodimeric complexes in macaques and comparison to single-chain IL-15 (P6242)
Authors: Pavlakis George , Bergamaschi Cristina , Valentin Antonio , Kulkarni Viraj , Bear Jenifer , Rosati Margherita , Alicea Candido , Sowder Raymond , Chertova Elena , Lifson Jeffrey , Felber Barbara 
Published: January 2023

Abstract
The common γ-chain cytokine IL-15 plays a significant role in CD8+ T and NK cell survival, proliferation and activation, and may find clinical applications in cancer. We have previously showed that co-expression of IL-15 and IL-15Rα in the same cell allows for efficient production of bioactive IL-15 heterodimer in vivo, whereas the single-chain IL-15 is unstable and poorly secreted. In addition, the identification of soluble IL-15/IL-15Rα complexes in human and mice sera suggests that the IL-15 heterodimer is the natural form of the cytokine in vivo. We have developed stable cell lines overproducing IL-15/IL-15Rα heterodimers. We compared pharmacokinetics and biological effects of purified IL-15 heterodimers with E.coli-derived single-chain IL-15 in macaques. Upon i.v. injection, the plasma half-life of IL-15 heterodimers was increased 6x compared to single-chain IL-15. Injection s.c. of IL-15 heterodimers resulted in persistent bioactive level of plasma IL-15 for up to 72 hrs. Repeated s.c. administration of IL-15 heterodimers resulted in a massive expansion of NK, γδ and CD8 T cells. Therefore, in comparison to single-chain IL-15, IL-15 heterodimer is more stable in vivo, has longer plasma half-life and is more bioactive in macaques. This favorable pharmacokinetics/pharmacodynamics profile allows for lower dose, simple s.c. delivery and lowers the possibility of toxicity due to cytokine spike, therefore it is the most favorable form for clinical applications of IL-15.
Air-pollution mediated susceptibility to inflammation and insulin resistance via CCR2 dependent and independent effects (P6246)
Authors: Liu Cuiqing , Xu Xiaohua , Bai Yuntao , Wang Tse-yao , Rao Xiaoquan , Sun Lixian , Ying Zhekang , Maiseyeu Andrei , Sun Qinghua , Rajagopalan Sanjay 
Published: January 2023

Abstract
Epidemiologic and experimental studies support an association between fine ambient particulate matter &lt;2.5µm (PM2.5) exposure and insulin resistance/type 2 diabetes mellitus. A role for innate immune cell activation has been suggested in the pathogenesis of these effects. We evaluated the role of CCR2 in PM2.5-mediated inflammation and insulin resistance. Wild-type C57BL/6 and CCR2-/- male mice were fed a high-fat diet and assigned to concentrated ambient PM2.5 or filtered air for 17 weeks via a whole body exposure system. PM2.5 exposure resulted in whole body insulin resistance and increased hepatic lipid accumulation in the liver which was attenuated in CCR2-/- mice by inhibiting SREBP1c mediated transcriptional programming, decreasing fatty acid uptake and p38 (but not JNK/ERK) MAPK activity. CCR2-/- restored abnormal phosphorylation levels of AKT, AMPK in visceral adipose tissue and adipose tissue macrophage content. However, the impaired whole body glucose tolerance and reduced GLUT4 in skeletal muscle in response to PM2.5 was not corrected by CCR2 deficiency. CCR2 modulates PM2.5-mediated insulin resistance by regulating visceral adipose tissue inflammation and hepatic lipid metabolism. These findings provide new mechanistic links between air pollution mediated insulin resistance and associated abnormalities in lipid metabolism and adipose inflammation.
Characterization of CXCR3 expression on B cell subsets in chronic Hepatitis C patients and its relation to autoimmunity. (P6250)
Authors: Reyes Elane , Sugalski Julia , Khasnis Atul , Antony Don 
Published: January 2023

Abstract
Hepatitis C virus (HCV) patients have extrahepatic manifestations, such as cryoglobulins. Cryoglubilins are immunoglubulins that precipitate in the cold and contain IgM rheumatoid factor. Vasculitis is an autoimmune manifestation detected in a subset of HCV patients with cryoglobulins (1-5%). HCV patients with mixed cryoglobulinemia have enhanced levels of CXCL10 (a chemotactic ligand of CXCR3) in their plasma (Antonelli.2008) and CXCR3+ B cells are detected in the liver compartment (Mizouchi. 2010). We analyzed by flow cytometric analysis a defined cohort of HCV patients (Cryoglobulin and Rheumatoid factor state) and healthy controls for CXCR3 and CXCR4 expression on B cell subsets. The chemokine expression was compared with parameters of B cell phenotype such as activation state (CD86), and cell cycling(ki-67) . We found that mature activated B cells of HCV patients had a high expression of CXCR3 (75% in HCV vs. 20% in healthy controls).. The goal of this project is to define the potential role that CXCR3 plays in the progression of HCV towards autoimmunity by comparison with alteration in B cell parameters such as alterations of cycling, apoptosis and survival.
Direct actions of TSLP on CD4+ T cells are important for in vivo Th2-type responses (P6252)
Authors: West Erin , Rochman Yrina , Kashyap Mohit , Li Peng , Spolski Rosanne , Leonard Warren 
Published: January 2023

Abstract
TSLP is a pleiotropic cytokine that has broad effects in multiple T helper type 2 (Th2) driven immune responses, including in asthma, intestinal immunity, and tumor progression. Initially, TSLP was shown to indirectly influence CD4+ T cells through direct actions on dendritic cells, resulting in CD4+ T cell priming towards a Th2 polarized response. Subsequently, direct actions of TSLP on CD4+ T cells were also demonstrated, but these direct actions were not analyzed in vivo. Herein, we show that TSLP induces both Th2 (IL-4, 5, 13) and Th2-associated (IL-3, 9, 10 and 31) cytokines early after TCR activation. Interestingly, TSLP can increase IL-5 production independent of IL-2 and IL-4 signaling; however, production of the other Th2 cytokines was a least partially dependent on IL-2 and IL-4. Additionally, we demonstrated that direct actions of TSLP on CD4+ T cells were important for Th2 polarization and cytokine production in an in vivo mouse lung inflammation model. Lastly, we found that TSLP plays a role in the development of IL-9 producing cells during a mouse chronic asthma model. Taken together, these data indicate a role for critical direct actions of TSLP in Th2 responses in vivo and indicate strategies for manipulating Th2 responses, including in allergic disease.
Lithium treatment reduced microglia activation and inflammation after irradiation to the immature brain (P6256)
Authors: Zhu Changlian , Xie Cuicui , Zhou Kai , Blomgren Klas 
Published: January 2023

Abstract
To evaluate the effects of lithium on microglia activation and inflammation after irradiation to the immature brain, male rat pups were injected 2 mmol/kg lithium chloride i.p. on postnatal day 7 (P7), additional lithium injections, 1 mmol/kg, were administered at 24 h intervals. Pups were subjected to whole brain 6Gy irradiation on P11. The pups were sacrificed at 6h and 24h after IR. Microglia scattered in the brain can be detected by counting their numbers, their size, engulfment of cell debris or by the production of cytokines and chemokines. Microglia were stained using the marker Iba-1, revealing the presence of these cells throughout the brain under normal conditions, and apparently higher numbers and bigger sizes after IR, particularly in areas where cell death occurred. Quantification of Iba-1+ cells in the GCL of DG showed a significant increase after IR, but with a lower increase in the lithium-treated brains. The concentration of MCP-1, IL-1α, IL-1β, GRO/KC in the hippocampus were increased significantly at 6h after IR compared with non-irradiation control. Lithium treatment significantly inhibited the increase. There was no significantly difference with these cytokines/chemokines at 24h after IR between lithium and vehicle treated group. These data indicate that lithium can specifically reduce inflammation through GSK3-beta inhibition or modified microglia activation.
Targeted delivery of HMGB1 siRNA to macrophages and dendritic cells for sepsis treatment in humanized mouse model (P6223)
Authors: Ye Chunting , Choi Jang-Gi , Abraham Sojan , Shankar Premlata , Manjunath N. 
Published: January 2023

Abstract
Over expression cytokine by innate immune cells is thought to initiate multiple organ failure in murine models of sepsis. Whether human cytokine storm also plays a similar role is not clear. Here we show that human hematopoietic cells are required to induce sepsis-induced mortality following cecal ligation and puncture (CLP) in the severely immunodeficient NOD/SCID/IL2Rγ-/- mice and siRNA treatment to inhibit HMGB1 release by human macrophages and dendritic cells dramatically reduces sepsis-induced mortality. Following CLP, as compared to immunocompetent WT mice, NOD/SCID/IL2Rγ-/- mice did not show high levels of serum HMGB1 or murine proinflammatory cytokines and were relatively resistant to sepsis-induced mortality. In contrast, NOD/SCID/IL2Rγ-/- mice transplanted with human hematopoietic stem cells (humanized BLT mice) showed high serum levels of HMGB1 as well as multiple human, but not murine proinflammatory cytokines and uniformly succumbed, suggesting human cytokines are sufficient to induce organ failure in this model. Moreover, targeted delivery of HMGB1 siRNA to human macrophages and dendritic cells using a short AchR binding peptide (RVG-9R) effectively suppressed secretion of HMGB1, reduced the human cytokine storm, human lymphocyte apoptosis and rescued humanized mice from CLP-induced mortality. SiRNA treatment was also effective when started after the appearance of sepsis symptoms. These results show that: CLP in humanized mice provides a model to study human sepsis; HMGB1 siRNA might provide a treatment strategy for human sepsis; and RVG-9R provides a tool to deliver siRNA to human macrophages and dendritic cells that could potentially be used suppress a variety of human inflammatory diseases.
Structure function analysis of NOD2, ATP and muramyl-dipeptide interaction (P1272)
Authors: Davis Beckley , Mo Jinyao , Smith Dylan , Pollock Sarah , Duncan J. 
Published: January 2023

Abstract
Nucleotide binding and oligomerization domain containing protein 2 (NOD2) is an intracellular protein that is involved in the recognition of bacterial cell wall derived muramyl-dipeptide. Mutations in the gene encoding NOD2 are associated with inherited inflammatory disorders including Crohn’s disease and Blau syndrome. NOD2 is a member of the Nucelotide-binding domain and leucine-rich repeat- containing protein gene family (NLR). Nucleotide binding is thought to play a critical role in signaling by NLR family members. However, the molecular mechanisms underlying signal transduction by these proteins remains largely unknown. Mutations in the nucleotide-binding domain of NOD2 have been shown to alter its signal transduction properties in response to muramyl-dipeptide in cellular assays. Using purified recombinant protein, we demonstrate that NOD2 binds and hydrolyzes ATP. Additionally, we have found the purified recombinant protein is able to bind directly to muramyl-dipeptide and can associate with known NOD2 interacting proteins in vitro. Binding of NOD2 to muramyl-dipeptide and homo-oligomerization of NOD2 are enhanced by ATP binding, suggesting a model that involves binding of nucleotide, muramyl-dipeptide, and oligomerization into a signaling complex. These findings set the stage for further studies into the molecular mechanisms that underlie detection of muramyl-dipeptide and assembly of NOD2-containing signaling complexes.
STING an essential trigger of DNA-mediated innate immune signaling. (P1258)
Authors: Barber Glen 
Published: January 2023

Abstract
Cellular innate immune sensor pathways are essential for identifying invading pathogens and for initiating host defense responses. However, how cells recognize DNA based microbes and trigger innate immune gene activation remains to be fully resolved. Here, we demonstrate that STING, an endoplasmic reticulum associated transmembrane protein, acts as a receptor for cytosolic pathogen associated DNA. This event invoked STING trafficking from the ER through autophagy-related endosomes to promote the activation of interferon regulatory factors IRF- 3, 7 and NF-κB, which triggered the production of host defense genes. STING, predominantly found in endothelial cells, macrophages and dendritic cells, was found to be essential for protecting mice against lethal HSV-1 infection through production of type I interferon. However, while STING was found to be essential for host defense, chronic activation of STING was observed to be responsible for inflammatory disease in two murine model systems. For example, DNaseII-/- and Trex1-/- mice die prematurely due to self-DNA manifested cytokine production. Lethality was prevented in both models through loss of STING since cytosolic self-DNA failed to robustly trigger cytokine production through STING signaling pathways. Our data suggests that STING senses aberrant cytosolic DNA species and is responsible for initiating innate immune gene activation, though may play a key role in inflammation and autoimmune disease manifested through self-DNA.
Defining cellular characteristics of human IL-7Rαlow memory CD8+ T cells using a genome-wide DNA methylation analysis (P1432)
Authors: Yoo Seung-Ah , You Sungyong , Shin Min Sun , Lee Naeun , Kang Ki Soo , Hwang Daehee , Kang Insoo 
Published: January 2023

Abstract
DNA methylation is one of the predominant epigenetic mechanisms to propagate heritable epigenetic information. Recently, we identified two subsets of memory CD8+ T cells expressing high and low levels of IL-7Rα in humans. IL-7Rαlow memory CD8+ T cells have impaired proliferation and survival in response to TCR triggering and IL-7, respectively. Importantly, the expansion of IL-7Rαlow memory CD8+ T occurs in human aging and lupus, suggesting a potential role for this cell subset in immunosenescence and autoimmunity. However, the exact characteristics of this cell subset are yet to be defined. Of interest, IL-7Rα expression in T cells is tightly regulated by DNA methylation in the IL-7Rα gene promoter. Here, we investigated how DNA methylation contributes to determining the distinct characteristics and functions of IL-7Rαhigh and IL-7Rαlow memory CD8+ T cells using genome-wide DNA methylation profiling technique. Methylation profiling revealed genes that are differentially methylated between IL-7Rαhigh and IL-7Rαlow memory CD8+ T cells. In particular, we found that the most significantly hypomethylated genes in IL-7Rαlow memory CD8+ T cells are associated with chemotaxis (CX3CR1, CXCR1, CCL3, and CCL4). Furthermore, the hypomethylation of these genes directly correlated with increased gene expression in this cell subset. Our data suggest that DNA methylation can be an important mechanism involved in defining the characteristics of IL-7Rαlow memory CD8+ T cells.
Histone deacetylase 11 is an epigenetic regulator of cytotoxic T-lymphocyte effector function and memory formation (P1404)
Authors: Woods David , Woan Karrune , Wang Dapeng , Yu Yu , Powers John , Sahakian Eva , Cheng Fengdong , Wang Hongwei , Rock-Klotz Jennifer , Villagra Alejandro , Pinilla-Ibarz Javier , Yu Xue-Zhong , Sotomayor Eduardo 
Published: January 2023

Abstract
The roles of individual histone deacetylases in the regulation of T-cell development and function remain largely unknown. Here we provide evidence for HDAC11 as an epigenetic regulator of T-cell inflammatory response as well as CTL central memory formation. To investigate the role of HDAC11 in T-cells, an HDAC11 knockout (HDAC11KO) mouse model was utilized. HDAC11KO mice display no gross phenotypic abnormalities and no alterations in T-cell development or CD4+/CD8+ population distributions. However, HDAC11KO CTLs are hyper-proliferative and secrete significantly higher levels of IL-2, TNF, and IFN-γ upon activation, and HDAC11KO mice accumulate higher percentages of central memory CTLs. In an allogeneic bone marrow transfer model, HDAC11KO T-cells mediate more potent and robust graft vs host disease associated with increased inflammatory cytokines. Mechanistically, we provide evidence that in CTLs HDAC11 interacts with the Eomes gene promoter, a known regulator of IFN-γ production and memory formation. HDAC11KO CTLs, at both basal state and post stimulation, display higher levels of acetylation at the Eomes promoter, indicative of a permissive transcriptional state. Correspondingly, eomes mRNA levels in HDAC11KO cells are elevated. Finally, chromatin immunoprecipitation of HDAC11 reveals interaction between HDAC11 and the Eomes promoter in CTLs. These results support HDAC11 as an epigenetic regulator of CTL function and memory formation through epigenetic regulation of Eomes.
T cell receptor repertoire plasticity in peripheral CD8 T cells (P1446)
Authors: Thomas Paul , Wang George , McClaren Jennifer , Morris Melissa , Doherty Peter , Dash Pradyot 
Published: January 2023

Abstract
Using a technique for the paired amplification of TCR α and β chains from single cells, we have discovered a surprising plasticity in the peripheral TCR repertoire, particularly under conditions of inflammation. This plasticity includes multiple instances of TCR revision, where changes are made in the TCR coding sequence by de novo rearrangement. Here we describe the induction of this process in murine CD8+ T cells derived from influenza-infected lungs and human T cells derived from CMV-infected individuals ex vivo or stimulated in single-cell cultures in vitro. Taking advantage of the co-expression of productive and non-productive Tcra alleles, we demonstrated secondary rearrangement in the TCRβ locus ex vivo, while the in vitro cultures demonstrated a remarkably high rate of revision following strong stimulation. Not surprisingly, revision was found to be RAG-dependent. Utilizing a model of peripheral deletion of RAG2, we showed that mice deficient in revision showed delayed onset and severity of disease in an experimental autoimmune encephalomyelitis (EAE) model, but also produced a less functional T cell response following infectious challenge. Thus, peripheral TCR revision appears to be a robust process that contributes to optimal T cell functionality during infection but may also contribute to the development of autoimmunity.
The intrinsic role of ROG in CD8+ T cell development has an impact on protective immunity against viral infection (P1452)
Authors: Shin Hyun Mu , Kapoor Varun , Welsh Raymond , Berg Leslie 
Published: January 2023

Abstract
Repressor of GATA3 (ROG) is a member of the poxviruses and zinc finger family of transcriptional repressors, and is also known as ZBTB32. Several studies have suggested that ROG may regulate the activation and differentiation CD4+ and CD8+ T cells, and further, may also play a role in plasma cell differentiation. We have investigated the role of ROG in the development of anti-viral memory T cells. Upon acute infection with LCMV Armstrong, Rog-/- mouse accumulated higher proportions and numbers of virus-specific CD8+ T cells, resulting in an increased number of memory cells. Rog-/- mouse exhibited normal viral clearance, but as a consequence of the elevated primary response, the Rog-/- CD8+ T cell population contains enhanced protective memory potential. We also examined the response of Rog-/- mice to infection with a high dose of LCMV-clone 13, which causes a chronic infection leading to exhaustion of the CD8+ T cells. We found that approximately one-half of Rog-/- mice infected with high dose LCMV-clone 13 succumbed to this infection as early as day 9 post-infection and also exhibited severe immune pathology in the lung compared to WT mice. The higher frequency of IFNg-producing virus-specific CD8+ T cells in Rog-/- mice also resulted in enhanced viral control following LCMV-clone 13 infection. These data indicate that ROG plays a unique non-redundant role during acute and chronic LCMV infection, and is an important regulator of CD8+ effector and memory T cell generation.