bluuebunny/crossref_metadata_2025_split huggingface dataset:
Muramyl dipeptide potentiates lipoteichoic acid-induced nitric oxide production in macrophage (135.32)
Authors: Jeon Jun-Ho , Yang Jihyun , Im Jintaek , Han Ji Young , Kim Sun Kyung , Baik Jung Eun , Yun Cheol-Heui , Han Seung Hyun 
Published: January 2023

Abstract
Pathogen-associated molecular patterns, such as lipoteichoic acid (LTA) and peptidoglycan (PGN) of gram-positive bacteria, often cooperate together to induce immune responses in the host. In this study, we investigated nitric oxide (NO) production by macrophages in response to LTA and muramyl dipeptide (MDP), a minimal structural unit of PGN. MDP alone could not induce NO production in both murine macrophages and a murine macrophage-like cell line, RAW 264.7, whereas LTA induced NO production. However, MDP potentiated LTA-induced NO production under the same condition. This was due to enhanced activation of Toll-like receptor 2 (TLR2) recognizing LTA since MDP increased TLR2 expression on the surface of macrophages and the potentiation effect was abolished in the macrophages derived from TLR2-deficient mice. Furthermore, MyD88 and MAP kinases such as ERK and p38 kinase also appear to be essentially required for such potentiation, which was determined by using MyD88-deficient macrophages and their specific inhibitors, PD98059 and SB203580, respectively. Subsequently, MDP enhanced the LTA-induced activation of NF-κB in RAW 264.7. These findings suggest that MDP potentiates the ability of LTA to stimulate macrophages via upregulating TLR2 activity. Supported by a grant from 21C Frontier Microbial Genomics and Application Center Program, Ministry of Science and Technology (11-2008-15-005-01).
The Pseudosubstrate domain of Protein Kinase C-epsilon directs translocation to phagosomes via binding to phosphomonoinositides (135.46)
Authors: Lennartz Michelle R. , Cheeseman Keylon , Hanes Cheryl M , Kashiwagi Kaori , Shirai Yasuhito , Saito Naoaki , Loegering Daniel J. 
Published: January 2023

Abstract
Protein kinase C-epsilon (PKC-ε) translocation is required for efficient IgG-mediated phagocytosis. We have published that the C1B region, binding to DAG generated by PLCγ1, is necessary for translocation (Cheeseman et al, MCB, 2006). However, C1B is not sufficient for translocation. Deletion and mutational analysis, as well as the use of PKC-ε/PKC-δ chimeras, indicate that the pseudosubstrate domain (εPS) is necessary for PKC-ε translocation. A PKC-ε fragment containing only the C1 and PS regions, as well as a chimera of PKC-δ containing εPS and εC1B, accumulated, demonstrating that these two regions are necessary and sufficient for targeting PKC-ε to phagosomes. Protein-lipid overlays of εPS-GST indicate that εPS preferentially associates with phosphomonoinositides (PI5P, PI3P&gt;PI4P&gt;&gt;&gt;polyphospho-inositides). Of the PI3 Kinase inhibitors, 100 nM wortmannin, but not 50 µM LY294002 inhibits PKC-ε translocation to phagosomes. That both drugs block the formation of PI3P as indicated by 2XFYVE staining argues against PI3P as the docking lipid for PKC-ε. Similar to PKC-ε, wortmannin, but not LY294002, blocked accumulation of PLCδPH (a PI4,5P2 reporter) at forming phagosomes. Together, these data suggest that εPS binding to PI4P and/or PI5P is necessary for PKC-ε translocation to phagosomes. Efforts are underway to identify the specific lipid binding partner for PKC-ε. Funded by NIH R01 AI05821 (MRL).
Chicken scavenger receptors and their ligand-induced cellular immune responses (135.57)
Authors: He Haiqi , MacKinnon Kathryn M. , Genovese Kenneth J. , Nerren Jessica R. , Swaggerty Christi L. , Kogut Michael H. 
Published: January 2023

Abstract
Scavenger receptors (SR) play an important role in the innate immunity. To evaluate the role of SR in the chicken immune system, we investigated immune responses, including oxidative burst, degranulation, phagocytosis, nitric oxide (NO) production, and cytokine gene expression of chicken heterophils, peripheral blood mononuclear cells, and a chicken macrophage cell line (HD11) to stimulations with various SR class A and B ligands. Our results show that ligands for SR-A, fucoidan, poly (I) and poly (G), but not ligands for SR-B, phosphatidylserine (PdS) and native LDL, stimulated a dose-dependent NO production in HD11 cells. However, these SR-A ligands failed to induce NO in monocytes isolated from peripheral blood. Quantitative RT-PCR indicated that SR ligands differentially affected the gene expression of several cytokines and chemokine in HD11 cells. SR-A ligands induced a strong up-regulation of proinflammatory cytokine IL-1β and to a lesser extent, IL-6, IL-12 and Mip-1β, with down-regulation of IL-4. However, SR-B ligands did not alter expression of these genes. Heterophil degranulation was not induced by any SR ligands. Stimulation of heterophils and monocytes with SR ligands also did not increase oxidative burst, with the exception of native LDL. However, LDL induced oxidative burst is probably mediated by the LDL receptor, not by SR.
MOLECULAR LINK BETWEEN TOLL LIKE RECEPTORS, TUMOROGENESIS AND OBESITY (135.61)
Authors: Kumaraguru Uday , Nandakumar Subhadra 
Published: January 2023

Abstract
Hygiene hypothesis states that "Lack of early childhood exposure to infectious agents and symbiotic microbes increases the susceptibility to allergic diseases by modulating the immune system". We predicted that neonate's exposure to microbes or its components will have long term protective effect against infectious diseases and cancer. To this end, we simulated dirty environment (exposure to microbial components) for a group of neonatal mice and compared them to mice kept in sterile conditions. The mice were allowed to grow up to 5-6 weeks after which they were analyzed for their ability to resist infection and tumor challenge. The mice that were in a simulated dirty environment were better able to resists HSV disease manifestation and tumor challenge with adeno carcinoma cells. Interestingly, the mice kept in sterile conditions gained significant body weight. Taken together, mice in cleaner environment became obese and were prone to infection and cancer indicating a similarity with human epidemiological data. We found that microbial exposure resulted in the 2.5 times more bone morphogenetic protein -7 in microbes exposed mice compared to mice kept in cleaner environment. BMP-7 is involved in the generation of brown fat that is known to be efficient in energy utilization, metabolism and weight reduction. Hence we conclude that bmp-7 induced by exposure to microbial components may be the molecular link between the microbial exposure pattern, obesity development and immune-protection. Increasing the levels of BMP-7 may well be employed as a therapeutic tool.
Acknowledgements:
ETSU-College of Medicine-Start up funds
Pathogen-induced activation of inflammasomes requires intracellular redistribution of the apoptosis associated speck-like protein containing a caspase recruitment domain (ASC) (135.70)
Authors: Bryan Nicole B , Dorfleutner Andrea , Rojanasakul Yon , Stehlik Christian 
Published: January 2023

Abstract
Activation of caspase-1, which is necessary for the maturation and secretion of the pro-inflammatory cytokines IL-1β and IL-18, occurs upon assembly of multi-protein complexes known as inflammasomes. Apoptosis associated speck-like protein containing a caspase recruitment domain (ASC) is an adaptor protein, which is essential for the recruitment of caspase-1 into inflammasomes. However, distribution of endogenous ASC has not been thoroughly examined. In the current study, we used laser scanning confocal microscopy and subcellular fractionation to demonstrate that endogenous ASC localized primarily to the nucleus in resting human macrophages. Upon pathogenic stimulation, ASC rapidly redistributed to the cytosol, and subsequently formed a characteristic perinuclear aggregate. These aggregates were found to incorporate other key inflammasome proteins including NLRP3 and caspase-1. Furthermore, sequestering ASC to the nucleus impaired formation of the inflammasome and abolished caspase-1 activation, as measured by IL-1β secretion. Our results indicate that the redistribution of ASC from the nucleus to the cytosol represents a novel and essential mechanism that regulates inflammasome assembly and activation.
Nod1 and Nod2 regulation of pulmonary immunity to Legionella pneumophila (135.77)
Authors: Berrington William Richard , Smith Kelley D , Wells Rick D , Jansson-Hutson Malinka , Iyer Ravi , Skerrett Shawn J , Hawn Thomas R 
Published: January 2023

Abstract
The role of Nod1 and Nod2 in pulmonary innate immune responses is poorly understood. We hypothesized that Nod1 and Nod2 regulate non-redundant and specific immune responses during murine in vivo infection with Legionella pneumophila (Lp). We first examined whether Nod1 and Nod2 regulate detection of Lp in HEK293 cells transfected with luciferase reporter constructs and either Nod1 or Nod2. Heat-killed Lp stimulated NF-κB and interferon-β-dependent promoter activity in cells transfected with either Nod1 or Nod2. To examine the role of Nod1 and Nod2 during in vivo infection, we exposed Nod1-/- and Nod2-/- and C57Bl/6 control mice to aerosolized Lp. Bronchoalveolar lavage fluid from Nod1-/- animals had decreased neutrophils compared to wild type animals at 4 and 24 hours post-infection. Furthermore, monocyte recruitment was also impaired at 24 hours. In addition Nod1-/- mice had increased lung bacterial loads at 72 hours in comparison to wild type and Nod2-/- mice. In contrast, there was no reproducible difference in cytokines in Nod1-/- lung homogenates at 4 hours and 24 hours. Conversely, Nod2-/- mice had impaired neutrophil cell recruitment at 24 hours, and only a trend to increased organism burden at 72 hours (p=0.07). Nod2-/- animals, however, showed reproducible impairment in cytokine production from lung homogenates. Lung histologic sections at later time points (3-7 days) had no detectable differences in inflammation in Nod1-/- or Nod2-/- mice in comparison to wild type mice. Overall these data indicate that Nod1 regulates early inflammatory cell recruitment of both neutrophils and monocytes to the alveolar space without detectable differences in inflammatory cytokine productions or changes in lung histology at later time points.
Catecholamine mediated S49 T lymphoma apoptosis is regulated by p38 MAPK (136.15)
Authors: LaJevic Melissa D , Suleiman Samia , Cohen Rhonna L , Chambers Donald A. 
Published: January 2023

Abstract
p38 MAPK is an important intracellular signaling mediator for a variety of cellular and environmental stressors and plays a role in cell survival, apoptosis, and inflammation. Previously, our lab has shown that norepinephrine (NE) treatment of S49 T lymphoma cells leads to increased activity of p38 MAPK by the classical βAR/AC/cAMP/PKA pathway. Exogenous cAMP or βAR stimulation of S49 cells leads to apoptosis. To study the relationship between p38 MAPK activity and apoptosis, S49 cells were incubated in the presence and absence of a cAMP analogue (8-bromo) with or without the p38 MAPK inhibitor SB203580. After 72 hours of cAMP treatment, S49 cell viability decreased by 50%, yet in the presence of SB203580 cell viability decreased by 35%. The steady state levels of FasL mRNA (an early biochemical marker of apoptosis) were analyzed after two hours of S49 cell treatment with 8-bromo cAMP in the absence or presence of SB203580. FasL mRNA levels increased 9.8 fold with 8-bromo cAMP treatment for 2 hours and was reversed 50% by p38 MAPK inhibition. These results suggest that activation of p38 MAPK by NE or cAMP is associated with T lymphoma cell apoptosis. Supported by NIH
Deletion of miRNAs in bone morrow disrupts iNKT cell development and function (136.19)
Authors: Zhou Li , Seo Kook-Heon , He Hong-Zhi , Pacholczyk Rafal , Meng Dong-Mei , Dong Zhen , Mi Qing-Sheng 
Published: January 2023

Abstract
MicroRNAs (miRNAs), a class of non-coding small RNAs, function as endogenous translation repressors of protein-coding genes. Accumulated evidences indicate that miRNAs are differentially regulated in developmental hematopoietic organs during hematopoietic lineage differentiation. Invariant Natural Killer T (iNKT) cells are potent regulators of diverse immune responses. The role of miRNAs in iNKT cell development is unknown. Here we test if lack of miRNAs following the deletion of Dicer, the miRNAs-processing enzyme, will affect iNKT cell development and function. We generated a mouse strain with tissue-specific disruption of Dicer in the hematopoietic progenitors, which is designated as Dicerfl/flTie2Cre+ (Dicer KO). Significantly decreased iNKT cell ratio and munber were found in the thymus, spleen, and liver of Dicer KO mice compared to littermate control (Dicerfl/flTie2cre-). The frequencies of mature CD44hiNK-1.1+ iNKT cells were significantly decreased in Dicer KO mice, suggesting that miRNAs may regulate iNKT cell maturation at the CD44low to CD44hi and NK1.1- to NK1.1+ checkpoints. Furthermore, miRNA-deficient peripheral iNKT cells display profound defects in alpha-GalCer-induced activation and cytokine production. Bone marrow (BM) from Dicer KO mice poorly reconstituted iNKT cells compared to BM from WT control mice, while thymic iNKT cells from WT mice poorly migrate to liver and spleen in irradiated Dicer KO mice compared to that in WT control mice. In summary, miRNAs are potent regulators of iNKT cell development, migration, and function. (Supported by JDRF).
MicroRNA-30c is down-regulated during NKL cell activation and cytotoxicity induced by CD226 crosslinking (136.20)
Authors: Chen Lihua , Gong Jiuyu , Wang Tao , Xu Zhuwei , Zhuang Ran , Zhang Yun , Yang Angang , Jin Boquan 
Published: January 2023

Abstract
MicroRNAs have been implicated as negative regulators controlling diverse biological processes at the level of posttranscriptional repression. There have been little report on NK cell associated miRs until now. CD226 is a kind of activation receptors located on NK cells and involved in the activation and cytotoxixity of NK cells. Previous study showed that CD226 cross-linking increased the CD107a expression on NKL cells. In the present study, we identified miRs induced in NKL cells after exposure to CD226 crosslinking using microarray and quantitive RT-PCR technology. It has been found that miR-30c level was down-regulated in NKL cells after CD226 crosslinking. Furthermore transfection of miR-30c into NKL cells decreased the CD107a expression on NKL cells induced by CD226 crosslinking, suggesting that miR-30c plays a role in NKL activation and cytotoxicity induced by CD226 crosslinking. Meanwhile miR-30c was predicted to base-pair with sequences in the 3' UTRs of the HMBOX gene. Altogether, our studies propose that miR-30c may play a role in control of CD226 signalling in NKL cells through a negative feedback regulation loop involving in down-regulation of HMBOX expression.
C8orf13/BLK association with systemic lupus erythematosus across different ethnicities (136.28)
Authors: Lu Rufei , Dominguez N , Lessard C , Deshmukh H , Vidal G , Macwana S , Kelly J , Kim X , Cobb B , Kaufman K , Bae S-C , Tsao B , Shen N , Langefeld C , Niewold T , Gilkeson G , Merrill J , Moser K , Harley J , Nath S , Gaffney P , James J , Guthridge J 
Published: January 2023

Abstract
SLE is a complex autoimmune disease with immunological abnormalities that targets B cell functions and development. Previous genome-wide association studies identified C8orf13/BLK as a novel SLE risk factor. BLK (B lymphoid tyrosine kinase) has been shown to have potential effects on B cell develoment. The main objectives of this study are to: 1.) assess this association in independent populations and 2.) determine if BLK risk alleles correlate with alteration in peripheral blood B cell development and protein expression. We genotyped 52 SNPs within the C8orf13/BLK region in European-derived, Asian and African American populations. rs13277113 is associated with SLE in all three populations. A few novel upstream SNPs within the intergenic region demonstrate a stronger association in Asians. Flow cytometry data suggests there is a significant difference in BLK expression and T2 cells B cell numbers between high-risk and non-risk individuals based on the risk haplotype. Our results suggest that the BLK genetic polymorphisms alter immune system development and function to increase the risk for SLE development. Supported by NIH RR020143, RR015577, NIAID-DAIT-BAA-05-11, AI031584, AR053483, AR48940, AI063274, AR052125, AR043247 and OCAST # HR08-037.
Determining the copy number variation (CNV) of human Complement Factor H Related Genes: from Southern blots to Real-time PCRs (136.30)
Authors: Kitzmiller Kathryn J , Yang Yan , Wu Yee Ling , Zhou Bi , Chung Erwin K , Yu C. Yung 
Published: January 2023

Abstract
Current applications for detecting the CNV of Complement Factor H-Related Genes 3 and 1, CFHR3 and CFHR1, such as Multiplex Ligation-Dependent Probe Amplification, MLPA, rely on the presence of single nucleotide polymorphisms that associate with but do not necessarily reflect the actual physical variation of gene copy numbers. A 76-kb "deletion" of genomic region for CFHR3-CFHR1was detectable using long range-mapping of PmeI-digested DNA resolved by Pulsed-Field Electrophoresis using a CFHR1 cDNA probe. The deletion was further characterized by Southern blots of genomic DNA digested by TaqI using CFHR2 cDNA probe, or PshAI-PvuII using CFH exons 21-22 probe. Thus, we developed quantitative Real-time PCR (RT-PCR) assays to detect the deleted region using 36 samples of genomic DNA characterized by Southern blot for CNV of CFHR3-CFHR1. Three RT-PCR assays were designed to measure a common region between CFHR3 and CFHR4, and CFHR1 and CFHR2, as well as intergenic region between CFHR3 and CFHR1using the standard curve method and obtaining the ratio between the target gene(s) and endogenous control. The 3 RT-PCR assays were able to correctly identify the CNV of CFHR3-R1 in 35/36 samples, for 97.5% accuracy and 100% sensitivity. This method will enable further investigation of CFHR3-R1CNVs in Age-Related Macular Degeneration, atypical Hemolytic Anemia and Systemic Lupus Erythematosus.
Epigenetic regulation of BCL6 and other genes important for B-cell maturation (136.32)
Authors: Ramachandrareddy Himabindu , Shen Yulei , Chan Wing C , McKeithan Timothy W 
Published: January 2023

Abstract
BCL6, a proto-oncogene frequently mutated and translocated in B-cell lymphoma, encodes a transcriptional repressor essential for the germinal center (GC) reaction. BCL6 mRNA and protein have higher expression in GC-B cells than in pre- and post-GC-B cells despite strong autorepression. Hypothesizing that histone modifications epigenetically regulate BCL6 during B-cell differentiation, we mapped a number of histone modifications associated with active transcription far upstream to the promoters of BCL6 and ~100 other genes in pre-GC-B, GC-B and plasma cell lines at nucleosomal resolution using the ChIP-chip assay. We identified numerous regions of histone H3 acetylation and H3 lysine-4 mono- and trimethylation (H3K4me1 and -3) within and upstream of BCL6; most striking was a vast region of H3K4me1 extending 140-260 kb upstream of BCL6 seen in a GC-B-cell line, but not in pre- or post-GC-B-cells. We also observed high expression of an intergenic transcript within this far upstream region in GC-B-cell lines but not in numerous other lines, suggesting distant regulatory regions, active only in GC-B-cells, account for high BCL6 expression. Many other genes also showed evidence for stage-specific distal regulatory regions. Defining a histone code for B cell master regulators will elucidate transcriptional networks central to both the immune response and cancer. The work is supported by UNMC Eppley Cancer Institute.
Identification of corticosteroid regulated genes in human Th2 cells (136.37)
Authors: Bowen Holly , Lee Tak H , Cousins David J , Lavender Paul 
Published: January 2023

Abstract
Glucocorticoids (GCs) are the mainstay treatment for many inflammatory diseases including asthma, which in the majority of patients can be controlled by inhaled GC. The molecular mechanisms of this regulation are however still not fully understood, especially in the small percentage of corticosteroid insensitive patients, who fail to respond well to even high doses of GCs.
In order to gain a deeper understanding of the molecular mechanisms of GC mediated gene regulation we have performed microarrays on in vitro differentiated Th1 & Th2 cells treated with the synthetic GC Dexamethasone (Dex). Naïve human CD4+ T cells were cultured for 28 days under either Th1 or Th2 skewing conditions. At day 28 these highly polarised cells (&gt;95% Th1 cells are IFNγ positive, and &gt;50% Th2 cells are interleukin (IL)-5 positive) were activated in the presence or absence of Dex at 10-7M. The isolated total RNA was used to probe the U133Plus2 gene chip (n=2), and results were confirmed via quantitative real-time PCR (n=4).
Our results have identified 64 genes consistently altered &gt; 2-fold by Dex treatment in differentiated human T cells. These genes include documented Dex targets such as GILZ and FKBP51, as well as some novel targets. We are currently identifying the pathways that these novel targets fall into, as well as investigating potential differences between to corticosteroid sensitive and insensitive asthmatic patients.
This work is funded by the MRC and Asthma UK.
Antisense transcripts negatively regulate transcription of multiple variegated killer immunoglobulin-like receptor (KIR) genes (136.38)
Authors: Cichocki Frank , Lenvik Todd , Anderson Stephen K , Miller Jeffrey S 
Published: January 2023

Abstract
The objective of this study is to understand the epigenetic mechanisms that regulate Killer Immunoglobulin-Like Receptor (KIR) gene regulation in human natural killer (NK) cells. We have previously identified both bidirectional transcription within the previously characterized KIR promoter and a novel promoter element upstream of the previously characterized promoter. Based on these findings, we propose that antisense transcripts homologous to the KIR promoter region participate in the establishment of the epigenetic modifications that silence individual KIR genes during human NK cell development. To test this prediction, we overexpressed KIR3DL1 antisense transcripts in developing NK cells and found that mature cells from these cultures exhibited a four-fold decrease in KIR expression relative to GFP-expressing controls. We have also identified 324 base pair and 28 base pair double-stranded RNA molecules homologous to the KIR3DL1 promoter region, suggesting that a siRNA mechanism may be responsible for establishment of the promoter methylation patterns within the KIR locus. Understanding the mechanisms of KIR expression, a requisite for NK cell education/licensing, may allow us to understand the acquisition of effector function and manipulate it for therapeutic benefit.
R01 HL 55417