bluuebunny/crossref_metadata_2025_split huggingface dataset:
Non-steroidal anti-inflammatory drugs blunt antibody production in human B lymphocytes from older volunteer (84.10)
Authors: Bancos Simona , Phipps Richard 
Published: January 2023

Abstract
Older individuals respond poorly to vaccination. B lymphocytes are responsible for the synthesis of antibodies that are generated in response to infection and vaccination. Our laboratory has shown that activated human B cells isolated from younger subjects (18-35 years old) express cyclooxygenase-2 (Cox-2) and that inhibition of Cox-2 by non-steroidal anti-inflammatory drugs (NSAIDs) blunts antibody production. However, NSAIDs are highly used by people over 60 years old. There is no information regarding Cox-2 expression in B cells in older human subjects (over 60 years old). We hypothesize that Cox-2 activity is reduced in B cells from older individuals. Young (18-35 years old) and older subjects (over 60 years old) were enrolled in the study. B cells were isolated from peripheral blood, stimulated in vitro with anti-Ig plus ODN CpG 2395 in the presence or absence of NSAIDs and Cox-2 expression, prostaglandin production, cytokines and antibody synthesis determined. Cell viability and proliferation were also assayed. Compared to younger subjects, B cells from older volunteers had reduced Cox-2 expression and also produced less antibody. Furthermore, in these individuals, antibody levels were further decreased by NSAIDs. Thus, NSAIDs and low intrinsic Cox-2 expression could be responsible for the poor antibody response to vaccination in the elderly.
Do Maternal Antibodies Inhibit Newborn Immunization? (84.15)
Authors: Akpinarli Akgul , Matzinger Polly 
Published: January 2023

Abstract
Investigations with measles vaccine suggested that persisting maternal antibodies might inhibit active immunization in the newborn. Because evolutionarily this makes no sense, it prompted us to revisit the relationship between maternal antibodies and neonatal immunity. We primed and boosted adult B6 mice against OVA and analyzed the level of OVA-specific antibodies in the serum of the progeny at various times after birth. The offspring of immunized mothers acquired high levels of antibodies, which lasted for the duration of lactation and then waned, disappearing at about 120 days for IgG1 and 50 days for IgG2a. Next, newborn mice were immunized after acquiring high titer of passively transferred antibody from their mother. We found that there was no apparent impairment of responsiveness as they made high titers of anti-OVA antibodies, and these did not wane, unlike those of their unimmunized littermates. Because the contribution of antibodies from the newborn could not be determined, we used a congenic marker system to discriminate maternal antibodies from those made by their progeny (Igha and Ighb). We found that newborn mice from both groups produced similar levels of Igha or Ighb antibodies against OVA. Therefore we conclude that newborn born immunization was not hindered by the presence of maternal antibodies.
Nocodazole induces apoptosis in human Jurkat T cells through G2/M arrest-dependent phosphorylation of Bcl-2 and Bim by p34cdc2 kinase and subsequent provoking mitochondrial death signaling pathway (85.16)
Authors: Kim Young Ho , Han Cho Rong , Jun Do Youn , Park Hae Sun , Woo Hyun Ju 
Published: January 2023

Abstract
Treatment of human Jurkat T cells with nocodazole, a microtubule-depolymerizing agent, provoked apoptosis along with G2/M-arrest, Bcl-2 phosphorylation at Thr-56 and Ser-70 residues, Bim phosphorylation, Δψm loss, and caspase cascade activation. Whereas these nocodazole-induced apoptotic events were abrogated by Bcl-2 overexpression, the G2/M-arrest and phosphorylation of Bcl-2 and Bim were sustained. Although Bcl-2 and Bim were mainly located in mitochondrial fraction regardless of their phosphorylation status, their physical interaction became reduced upon phosphorylation. FADD-positive Jurkat clone A3 and FADD-deficient Jurkat clone I2.1 exhibited similar susceptibilities to the apoptogenic activity of nocodazole. Further analysis using selective caspase inhibitors elucidated that the activation of caspase-9 and -3, leading to activation of caspase-7 and -8, was crucial for nocodazole-induced apoptosis. In the presence of the G1/S blocking agent (hydroxyurea), nocodazole failed to induce G2/M-arrest, phosphorylation of Bcl-2 and Bim, and mitochondria-dependent apoptotic events. Nocodazole-induced phosphorylation of Bcl-2 and Bim, Δψm loss, and apoptotic events were significantly reduced by pretreatment with p34cdc2 kinase inhibitor (roscovitine). Consequently, these results indicated that G2/M-arrest and resultant phosphorylation of Bcl-2 and Bim dictated by p34cdc2 kinase, contributed to mitochondria-dependent apoptosis in Jurkat T cells exposed to nocodazole.
Regulation of TRAIL Expression in T helper cells (85.17)
Authors: Zhang Jimin , Shi Yufang 
Published: January 2023

Abstract
TNF-related apoptosis-inducing ligand (TRAIL/Apo2 ligand), along with FasL and TNFα, is one of three members of the TNF cytokine family that induce apoptosis. TRAIL can bind to several different receptors including those containing a cytoplasmic death domain and others that lack such a domain, termed decoy receptors. Here we found that TRAIL was preferentially expressed in Th2 cells, and, though exogenous TRAIL did not cause death in these cells, it did induce apoptosis of Th1 cells. Since it differentially induced apoptosis in Th1 and Th2 cells, TRAIL was likely to modulate the balance between them in determining the type of immune response. However, the regulation mechanism of TRAIL expression in T lymphocytes is not fully understood. We showed that A1.1 T hybridoma cells constitutively expressed TRAIL at a low level that could be boosted by adding IFN-γ. When Th2 cells were treated with high dosage of IFN-γ upon restimulation, TRAIL expression was boosted as well, whereas when Th1 cells were treated with high dosage of IL-4 upon restimulation, TRAIL was surprisingly detected at a comparable level to that in Th2 cells without any cytokine treatment. Interestingly, although Th1 cells were the main producers of IFNγ, Th2 cells expressed IFNγ receptors. All these results suggest that IFN-γ secreted by Th1 cells and IL-4 secreted by Th2 cells may be potent inducers of TRAIL under certain circumstances and therefore can provide some insight into clinical applications.
Deciphering the function of the microRNA-181a in effector CD4+ T cells (85.19)
Authors: Deng Min , Arias Marco , Ramaswamy Madhu , Siegel Richard 
Published: January 2023

Abstract
One of the mechanisms of peripheral tolerance in CD4 T cells, mediated by the TNF superfamily member Fas/FasL, occurs through a process termed Restimulation Induced Cell Death. Previously, we showed that naïve and memory activated CD4 T cell subsets vary in their response to Fas apoptosis, with the effector memory subsets having the highest Fas sensitivity. Since this phenotype was maintained stably in activated cell cultures, genetic or epigenetic regulation is likely. To examine the role of miRNAs in regulating the phenotype of effector memory (Em) T cells, we performed a miRNA array via Solexa sequencing to profile miRNAs differentially expressed in CD4 T cell subsets. One of these, miR181a, was upregulated in naïve and Cm, but down modulated in memory and Em cells. This downregulation correlated with the upregulation of its target genes including DUSP6. Mir181a is required for positive selection of T cells in the thymus, however, its function in peripheral TCR signaling is unknown. To probe the functional consequences of differential miR181a expression in peripheral T cell subsets we utilized a siRNA knockdown and lentiviral overexpression approach. Thus far we have not observed overt differences in Fas sensitivity, however, we have seen skewing of cytokine responses in miR181a knockdown, including changes in IL-17 and IFN-g expression following TCR triggering. These results suggest a role for miR-181a in regulating certain aspects of the effector memory T cell phenotype.
Human pluripotent stem cells: a source of mast cells for the study of allergic and inflammatory diseases (86.16)
Authors: Kovarova Martina , Latour Anne , Chason Kelly , Tilley Stephen , Koller Beverly 
Published: January 2023

Abstract
Human mast cells are tissue resident cells with a principal role in allergic disorders. Cross-linking of the high-affinity receptor for IgE (FcϵRI) results in release of inflammatory mediators initiating the clinical symptoms of allergy and anaphylaxis. Much of our knowledge regarding the mechanisms of mast cell activation comes from studies of mouse bone marrow derived mast cells. However, clear differences have been identified between human and mouse mast cells. Studies of human mast cells are hampered by the limited sources available for their isolation, the resistance of these cells to genetic manipulation, and differences between cultures established from different individuals. To address this limitation, we developed a simple co-culture-free method for obtaining mast cells from human pluripotent stem cells (hPS) including human embryonic stem (hES) cells and induced pluripotent stem (iPS) cells. hES-derived mast cells respond to antigen by releasing mast cell mediators. Moreover, the cells can be generated in numbers sufficient for studies of the pathways involved in their effector functions. Genetically modified mast cells expressing GFP and Neo resistance were obtained by modification of hES cells prior to differentiation. iPS cells can be used for derivation of mast cells with patient specific genetic make-up. This direct co-culture-free differentiation of hPS cells represents a new and unique model to analyze the function and development of human mast cells.
Regulation of RGS13 expression by the tumor suppressor p53 in mast cells (86.22)
Authors: Druey Kirk , Lu Yunbiao , Kuehn Hyesun , Gilfillan Alasdair , Xie Zhihui 
Published: January 2023

Abstract
Regulators of G-protein Signaling (RGS) proteins negatively regulate GPCRs by accelerating the GTPase activity of Gα. In recent years, non-GPCR-related functions have also been described for these proteins. We found previously that RGS13 was enriched in mast cells and inhibited anaphylaxis in mice by suppressing IgE-mediated mast cell degranulation. We therefore examined the transcriptional regulation of RGS13 in mast cells. We confirmed the transcriptional start site (TSS) by 5’ RACE in cDNA from the human mast cell line LAD2 and demonstrated promoter activity in a ~ 1 kb element upstream of exon 1 using luciferase reporter assays. We also identified 2 potential p53 response elements (RE) in the region 2 kb upstream of the TSS by bioinformatic analysis. Overexpression and shRNA-mediated knockdown of endogenous p53 in LAD2 cells revealed an inverse relationship between p53 levels and RGS13 expression. Endogenous p53 in these cells bound exclusively to oligonucleotides containing the p53 RE most upstream of the TSS. Cells expressing double-stranded oligonucleotides containing this site or a p53 consensus site had enhanced promoter activity. These studies indicate that p53 suppresses RGS13 transcription in mast cells. Thus, upregulation of p53 activity in mast cells exposed to stress could reduce the threshold for degranulation as a result of decreased RGS13 expression.
Effects of a novel mucosal tolerogen on critical T cell cytokine secretion and development of anti-FVIII inhibitor Ab in a mouse model of Hemophilia A (87.27)
Authors: Csencsits-Smith Keri , Nguyen Trinh , Donnachie Elizabeth , Borck Margarete , Escobar Miguel , Pascual David 
Published: January 2023

Abstract
The development of anti-FVIII (F8) antibodies (Abs) in Hemophilia A patients who receive therapeutic supplemental F8 results in significant morbidity. T cell help is required for anti-F8 inhibitor formation, however, the role of secreted cytokines is not yet fully understood. To ascertain the T cell cytokines important in the development of inhibitor Abs, F8-deficient mice were immunized 4x with 2μg recombinant human F8 at weekly intervals. To abate the production of inhibitor Ab, mice were intranasally immunized with 100μg of E. coli colony-forming antigen I (CFA/I), a novel tolerogen which reduces inflammatory immune responses. Plasma was assayed for the presence of secreted cytokines via multiplex and for functional anti-F8 Ab via Bethesda assay. Inhibitor Ab was detectable by day 14 post-primary immunization, and Th1 (IFNγ and IL-12), Th2 (IL-4), and Th17 (IL-17) cytokines were present in control and CFA/I treated plasma as early as 7 days post-primary immunization. There was no apparent correlation between Th1 vs Th2 responses and increased inhibitor titer. However, mice that were resistant to the effects of CFA/I treatment displayed increased anti-F8 Ab as well as IL-6 and IL-17 by day 21 post-primary immunization. In contrast, IL-13 secretion was associated with significantly decreased inhibitor antibody in CFA/I treated mice. Our results suggest that modification of Th cytokine response may amerliorate inhibitor Ab formation.
Adenosine A2A receptor activation inhibits T cell activity in a murine model of chronic granulomatous disease (87.3)
Authors: Lappas Courtney , Domeier Phillip , Chehata Veronica 
Published: January 2023

Abstract
Chronic granulomatous disease (CGD) is caused by defects in the NADPH oxidase system, and is characterized by an increased susceptibility to recurrent, severe bacterial and fungal infections. Early diagnosis and aggressive treatment can be effective at combating life threatening infections, but other significant complications of CGD, such as non-infectious hyperinflammatory manifestations, result in a high associated morbidity. Recent studies suggest that the hyperinflammation observed in CGD (gp91phox deficient) mice may be due in part to dysregulated T cell activity. We have broadly characterized the phenotypes of T lymphocytes collected from p91phox deficient mice and our data indicate that although the basal activation states of T cells collected from WT and CGD mice are similar, upon activation through the T cell receptor, CGD T cells produce higher levels of the pro-inflammatory cytokines IFN-γ and IL-2 and express elevated levels of the cell surface activation markers CD25, CD69 and CD40L as compared to WT T cells. This hyper-activation of CGD T cells is effectively modulated by treatment with the selective adenosine A2A receptor agonist CGS21680, which reduces the levels of IFN-γ and IL-2 by approximately 90% and down regulates CD25, CD69 and CD40L expression by approximately 50%. These data suggest that the adenosine A2A receptor is a potential pharmacological target for the treatment of the hyperinflammatory complications observed in CGD patients.
Identification of an XBP-1-Binding Endoplasmic Reticulum Stress-Responsive Enhancer of IFN-β Transcription in Macrophages (88.11)
Authors: Smith Judith , Liu Yi-Ping , Chen Hui , Qi Ling , Zeng Ling 
Published: January 2023

Abstract
Perturbation of the endoplasmic reticulum (ER) by various insults results in a conserved stress response called the “Unfolded Protein Response” (UPR). We have previously shown that macrophages undergoing a UPR respond to LPS with log-fold increased production of IFN-β a cytokine with diverse roles in innate and adaptive immunity. In this study, we found that pre-treatment of macrophages with the ER stress inducer thapsigargin resulted in augmented recruitment of IRF-3, CBP/p300, and ultimately transcriptional machinery, to the ifnb1 gene promoter during LPS stimulation. Although full synergistic IFN-β production required XBP-1, this UPR regulated transcription factor does not bind directly to the ifnb1 promoter. Analysis of the genomic DNA surrounding the ifnb1 gene revealed a conserved site 6.1kb downstream of ifnb1 that bound XBP-1 along with IRF-3, and CBP only during concomitant UPR and LPS stimulation. XBP-1 co-precipitates with p300, suggesting a mechanism of multi-molecular assembly. Results from luciferase reporter assays provide evidence that this +6kb region functions as an XBP-1-dependent enhancer of ifnb1 promoter activity. These findings provide a mechanistic link for diseases involving ER stress and type I interferon, including viral infection, ischemia-reperfusion injury, protein-misfolding and inflammatory diseases.
Distal regions of the human IFNG locus direct cell-type specific expression (88.12)
Authors: Collins Patrick , Henderson Melodie , Chang Shojing , Davis Georgia , McLoed Allyson , Mortlock Douglas , Aune Thomas 
Published: January 2023

Abstract
Genes, such as IFNG, which are expressed in multiple cell lineages of the immune system, may employ a common set of regulatory elements to direct transcription in multiple cell types or individual regulatory elements to direct expression in individual cell lineages. By employing a BAC transgenic system, we demonstrate that IFNG employs unique regulatory elements to achieve lineage specific transcriptional control. Specifically, a one 1-kb element 30 kb upstream of IFNG activates transcription in T cells and NKT cells but not NK cells, macrophages and dendritic cells. This distal regulatory element is a Runx3 binding site in Th1 cells, and is needed for RNA polymerase II recruitment to IFNG, but not for histone acetylation of the IFNG locus. These results support a model whereby IFNG utilizes distinct regulatory elements to achieve cell-type expression.
DNMT3a mediates lineage-specific, de novo DNA methylation at the ifng promoter and contributes to ifng gene silencing in Th2, Th17 and Treg cells (88.15)
Authors: Thomas Rajan , Gamper Christopher , Powell Jonathan , Wells Andrew 
Published: January 2023

Abstract
Naïve CD4+ T cells can differentiate into distinct subsets of effectors that express unique cytokine profiles upon activation. T helper 1 (Th1) cells robustly secrete IFNγ, a cytokine essential for clearance of intracellular pathogens and anti-tumor responses, while Th2, Th17, and regulatory T cells (Tregs) do not. The silencing of “off-pathway” cytokine genes during differentiation is incompletely understood, but may involve epigenetic mechanisms of gene repression. CpG dinucleotides in the ifng promoter, which are largely unmethylated in naïve CD4+ T cells and Th1 cells, are sites of substantial de novo methylation in non-IFNγ-producing lineages. To determine whether such de novo DNA methylation contributes to ifng gene silencing, we used mice with T cell-specific deletion of the de novo DNA methyltransferase, DNMT3a. While “on-pathway” cytokine expression polarized normally, DNMT3a-deficient CD4 T cells did not accumulate de novo ifng promoter methylation seen in wild-type cells during Th2, Th17 or Treg differentiation. Maintenance DNA methylation at the ifng intronic enhancer was not affected. Failed de novo methylation was associated with failed silencing, as DNMT3a-deficient Th2, Th17 and Treg cells produce significant levels of IFN-γ after secondary re-polarization toward the Th1 lineage. These results indicate that DNMT3a restricts the plasticity of CD4+ T helper cell fates via de novo DNA methylation of the ifng promoter in Th2, Th17 and Treg cells.
E2F1 Suppreses DC Maturation (88.22)
Authors: Xiao Weihua , Fang Fang , Wang Yan , Li Rui , Zhao Ying , Yang Guo , Jiang Ming , Sun Jie , Ma Yang , Ren Zijia , Tian Zhigang , Feng Wei , Yang De 
Published: January 2023

Abstract
E2F1 has been largely studied as a promoter of cell cycle and as regulator of apoptosis. Recently, E2F1 has been shown to regulate a wide range of genes in response to inflammatory stimulation of macrophages and to contribute to T cell activation in response to pathogens, implicating an extensive immunological role for E2F1. Dendritic cells (DCs) play critical roles as professional antigen-presenting cells in the development of immune responses, however, it is unclear whether E2F1 has any effect on DC phenotype and function. Here, we report that E2F1 acts as a suppressor of DC maturation. The level of E2F1 expression was transiently downregulated in the course of LPS-induced maturation of both human monocyte-derived dendritic cells and a mouse DC cell line, DC2.4. Knockdown of E2F1 by siRNA in DC2.4 cells resulted in both phenotypic and functional maturation, even without LPS treatment. Conversely, ectopic overexpression of E2F1 suppressed LPS-induced maturation of DC2.4 cells. Furthermore, knockdown of E2F1 caused the activation of several major signaling pathways known to be activated in the course of DC maturation, including Erk1/2, NF-κB and PI3K/Akt, suggesting that E2F1 may be involved in regulating multiple signaling pathways in DCs. Finally, the alternation by E2F1 on phenotypic maturation was confirmed with bone marrow-derived DCs from E2F1-KO mice. Overall, our data demonstrate for the first time that E2F1 is a critical regulator of dendritic cell maturation.
Role of Dnmt3a in cytokine gene regulation during Th1/Th2 differentiation (88.23)
Authors: Yu Qing , Kaplan Mark 
Published: January 2023

Abstract
Helper T cell polarization is accompanied by DNA methylation-mediated silencing of specific genes. Dnmt3a/3b, de novo DNA methyltransferases, have been shown to associate with cytokine gene promoters during Th1/Th2 differentiation. To elucidate the functional significance of Dnmt3a/3b-mediated DNA methylation in gene regulation of CD4 cell development, we generated conditional knockout mice by crossing Dnmt3aloxP/loxP and Dnmt3bloxP/loxP mice (a gift from Dr. En Li) with CD4-CRE mice. The conditional knockout mice exhibited normal lymphoid organ cellularity. Analysis of gene expression revealed that Dnmt3a-deficient Th1/Th2 cells secreted higher levels of effector cytokines in comparison with littermate controls, whereas absence of Dnmt3b did not show significant change, compared to controls. Using a DNA methylation-sensitive restriction method, we found that although absence of Dnmt3a did not alter global methylation levels of CD4 cells, Dnmt3a deficiency led to reduced levels of DNA methylation at specific gene loci, indicating that Dnmt3a participated in the establishment of local DNA methylation. Meanwhile, Chromatin IP data showed histone modification alterations in the absence of Dnmt3a. Taken together, the results indicated that Dnmt3a had an inhibitory effect on gene expression in Th1/Th2 development.
Arsenic enhances Langerhans cell migration to draining lymph nodes via CCL21 and IFN-γproduction in a model of epicutaneous protein sensitization (89.19)
Authors: Lee Chih-Hung , Hong Chien-Hui , Yu Chia-Li , Wang Li-Fang , Liao Wei-Ting , Huang Shau-Ku , Chen Gwo-Shin , Yu Hsin-Su 
Published: January 2023

Abstract
Arsenic induces multiple Bowen's disease (As-BD), or cutaneous carcinoma in situ, that runs an indolent course. Immune responses are important modifiers for the development of As-BD. We have previously reported that Langerhans cells (LCs) were reduced in As-BD, but how arsenic alters LC migration and polarizes Th responses remains unknown. Using an epicutaneous protein sensitization model, Balb/c mice were patch-sensitized with OVA and received feeding of 300 ppm arsenic or PBS for 200ul for 5 days. Defined as CD207+MHCII+ cells, the number of LCs was significantly decreased in patched epidermis, but increased in draining lymph nodes, with no changes in dermis and spleens 96 hours after OVA sensitization in arsenic-treated mice. There was a parallel increase of CCL21 in lymph nodes from arsenic-treated mice, and enhanced cellular proliferation and IFN-γ secretion in vitro were also noted in lymph node cells from arsenic-treated mice. Notably, in vivo administration of a neutralizing antibody against CCL21 abolished the increase of LCs in lymph nodes. These results suggest, therefore, that in this model, arsenic exposure enhanced LC migration to draining lymph nodes via CCL21 and interferon-γ production, and that chronic nature of As-BD might result from enhanced Th1 responses and LC migration.