bluuebunny/crossref_metadata_2025_split huggingface dataset:
The cellularity of thymic regulatory T cells is regulated by CCR7 through a non-cell intrinsic mechanism (HEM7P.228)
Authors: Hu Zicheng , Ehrlich Lauren 
Published: January 2023

Abstract
To ensure central tolerance, developing T cells must enter the thymic medulla to encounter self-antigens that induce autoreactive cells to undergo apoptosis or differentiation into the regulatory T cell (Treg) lineage. Both dendritic cells (DCs) and medullary thymic epithelial cells (mTECs) present antigens to tolerize thymocytes. mTECs express a diverse array of self-antigens, while DCs express auto-antigens acquired both in the periphery and from mTECs. Thymocytes must interact efficiently with these medullary antigen presenting cells (APCs) to encounter the full spectrum of self- antigens. CCR7 guides thymocytes into the medulla, promoting encounters with medullary DCs and mTECs. In the absence of CCR7, negative selection of autoreactive thymocytes is impaired, and autoimmunity ensues. Surprisingly, thymic Treg cellularity is increased in Ccr7-/- mice. Diminished negative selection has previously been associated with a concomitant increase in Treg differentiation. Thus, we anticipated that a cell-intrinsic defect in CCR7-/- thymocytes would account for increased Treg generation. Instead, a series of bone marrow chimeras demonstrate that CCR7-deficient non-T, non-B lineage hematopoietic cells account for increased Treg cellularity. Expression profiling revealed expression of Ccr7 by thymic DCs. We present evidence that CCR7 deficiency in thymic DCs is responsible for increased Treg cellularity, suggesting that CCR7 has pleiotropic effects on thymic central tolerance.
DNA-dependent protein kinase plays a role in inflammation independent of DNA-damage repair function and its inhibition blocks allergic inflammation in mice and modulates human CD4+ T cell function without causing SCID (HYP7P.264)
Authors: Pyakurel Kusma , Ghonim Mohamed , Ju Jihang , Rodriguez Paulo , Lammi Matthew , Davis Christian , Naura Amarjit , Boulares Hamid 
Published: January 2023

Abstract
We reported that DNA- dependent protein kinase (DNA-PK) is critical for the expression of NF-κB-dependent genes in TNF-α -treated glioblastoma cells including VCAM-1. Here we show that DNA-PK protein level and function are critical for VCAM-1 expression in TNF-α-treated human endothelial and mouse lung smooth muscle cells and are required for monocytes adhesion. Interestingly, DNA-PK activation and subsequent VCAM-1 expression in presence of TNF occurred independently of DNA breaks/repair. Administration of the DNA-PK inhibitor, NU7441, and DNA-PK heterozygosity reduced airway eosinophilia, mucus hypersecretion, airway hyperresponsiveness (AHR), and OVA-specific IgE production in asthma model of mice. Such effects correlated with a marked reduction in lung VCAM-1 expression and production of several inflammatory cytokines. Remarkably, such protection occurred without causing SCID. These results were confirmed in a chronic model of asthma using house dust mite. DNA-PK inhibition reduced Th2 cytokines production without affecting Th1 cytokines production and cell proliferation in CD3/CD28-stimulated human CD4+T cells potentially by blocking gata-3 expression. In mouse CD4+T cells, DNA-PK inhibition, in vitro, severely blocked CD3/CD28-induced gata-3 and t-bet expression and prevented differentiation of Th1 and Th2 cells. Thus, our results suggest DNA-PK as a novel determinant of asthma and a potential target for the treatment of the disease.
Monocyte and Treg phenotypes in two U.S. farming populations mirror differential asthma and atopy risk: studies in Amish and Hutterite children (HYP7P.254)
Authors: Hrusch Cara , Ober Carole , Stein Michelle , Igartua Catherine , Holbreich Mark , Thorne Peter , Vercelli Donata , von Mutius Erika , Sperling Anne 
Published: January 2023

Abstract
The differences in asthma prevalence between farming and non-farming families in central Europe are recapitulated in two unique U.S. farming populations, the Amish of Indiana and the Hutterites of South Dakota. The prevalence of asthma and allergic sensitization was 5.2% and 7.2%, respectively, in Amish children and &gt;15% and &gt;40%, respectively, in Hutterite children. Compared to Hutterite children, Amish children’s PBLs had lower eosinophils, but higher neutrophils. Amish had higher gene expression of IRF7 and TNFA, which may be the result of the elevated levels of endotoxin found in Amish homes. To determine how the Amish environment alters antigen presentation, we measured the activation status of monocytes. The Amish had increased percentages of inhibitory monocytes that expressed low HLA-DR and high ILT3 and ILT5. We investigated how these changes in innate activation influenced the adaptive immune response. Amish had low ICOS expression on their CD4 effector cells and high ICOS expression on their Tregs. The expression of ILT5 on monocytes correlated to the level of CD45RO+ICOS+ Tregs. Consistent with the idea that microbial exposure could be promoting a regulatory immune phenotype similar to endotoxin-induced tolerance, PBLs from the Amish were hyporesponsive to LPS. These results indicate that environmental exposure has a profound impact on baseline immune function and may prevent the induction of atopic disease in children.
Maternal supplementation of allergic female mice with gamma-tocopherol increases the development of select dendritic cell subsets and allergic lung inflammation in neonates (HYP7P.272)
Authors: Abdala-Valencia Hiam , Soveg Frank , Cook-Mills Joan 
Published: January 2023

Abstract
Offspring from allergic mothers have elevated numbers of CD11b+ lung dendritic cells and develop allergic lung responses to suboptimal allergen challenge. We recently reported that maternal supplementation with alpha-tocopherol (αT) reduced development of these dendritic cell subsets and allergic responses in offspring of allergic female mice. It is not known whether gamma-tocopherol (γT) supplementation regulates development of allergic responses. To address this, allergic female mice were supplemented with γT during pregnancy/lactation. Then, offspring were given a suboptimal allergen challenge. γT supplementation of allergic mothers elevated pup responses to allergen challenge. There were increased numbers of pup lung eosinophils, inflammatory mediators, and CD11b+ but not CD11b- subsets of CD11c+ dendritic cells. There were elevated numbers of IRF4+ CD11b+ CD11c+ dendritic cells, a dendritic cell subset critical for development of allergic responses. There were also fewer pups from γT supplemented allergic mothers. In conclusion, maternal αT supplementation reduced and maternal γT supplementation increased development of CD11b+ CD11c+ dendritic cells and allergic responses in offspring from allergic mothers. These results have implications for supplementation of allergic mothers with tocopherol isoforms and for development of allergies in future generations.
TGFb inhibits IL-33-mediated mast cell activation (INC6P.316)
Authors: Ndaw Victor , Paez Patrick , Taruselli Marcela , Ryan John 
Published: January 2023

Abstract
TGFβ is involved in many pathological considtions, including autoimmune disorders, cancer, and cardiovascular and allergic diseases. We have previously found that TGFβ can suppress IgE-mediated mast cell activation. IL-33 is a recently discovered member of the IL-1 family that plays a critical role in mast cell activation and enhances IgE-mediated activation. In this study, we investigated the effects of TGFβ on IL-33-mediated mast cell activation. Bone marrow-derived mast cells cultured in TGFβ -1, -2, or -3 showed reduced IL-33-mediated production of TNF, IL-6, IL-13 and MCP-1, in a concentration-dependent manner. Furthermore, TGFβ also reduced expression of the T1/ST2 receptor as well as IL-33-mediated ERK activation. The role of IL-33 in the pathogenesis of allergic diseases is incompletely understood. These findings, consistent with our previously reported effects of TGFβ on IgE-mediated activation, demonstrate that TGFβ can provide broad and substantial inhibitory signals to activated mast cells.
Generation and validation of a reporter mouse for inflammasome assembly (INM6P.335)
Authors: Tzeng Te-Chen 
Published: January 2023

Abstract
Inflammasomes are key signaling platforms that detect pathogenic microorganisms and sterile stressors and then activate the highly pro-inflammatory cytokines interleukin-1β (IL-1β) and IL-18 in response. Both IL-1β and IL-18 have numerous functions in inducing the production of more cytokines, such as TNFα. Hence, the inflammasome is a central mediator of inflammation. Although we have several tools to test inflammasome activation in vitro, there is a dearth of in vivo tools to probe the role of inflammasome activation during infection. Thus, we generated a transgenic mouse expressing mouse ASC-citrine fusion protein in the Rosa26 locus. This system contains a knock-in of ASC-citrine gene and a proximal floxed stop site, which allows us to conditionally express ASC-citrine in a lineage specific manner. BMDC from this reporter mouse showed ASC speck formation after inflammasome stimulation, as defined biochemically by others. This suggests that inflammasome activation will be detectable in vivo by looking for specks in tissues. Interestingly, our data suggested that non-hematopoietic cells are the major speck formed cells in the early phase of inflammation when infected mice with Influenza A virus. We also showed that specks are released after inflammasome activation that may enhance and amplify the inflammatory responses. Overall, the expression of ASC-citrine molecules in the mouse system has renders it a useful tool to study and monitor inflammasome activation in vivo.
B cell intrinsic effects of the vitamin D receptor on IgE responses. (IRC10P.417)
Authors: James Jamaal , Cantorna Margherita 
Published: January 2023

Abstract
B cells express the vitamin D receptor (VDR). VDR KO mice had higher levels of circulating total IgE than WT mice by 12 wks of age. Mice were sensitized using two OVA/alum injections. VDR KO mice mounted a significantly higher OVA-specific IgE response compared to WT mice. The data also showed that vitamin D deficient mice had higher OVA-specific IgE responses than vitamin D sufficient mice. IL-4, IL-5, IL-2, IFN-γ and IL-17 were not different in OVA re-stimulated spleen and mesenteric lymph nodes (MLN) in WT and VDR KO mice. However, VDR KO spleen and MLN produced significantly less IL-10 than WT. Furthermore B cells and not T cells were the major source of IL-10 in both the WT and VDR KO cultures. Experiments were done using T cell specific-VDR deletion (T-VDR KO), or B cell specific-VDR deletion (B-VDR KO). The serum IgE levels in T-VDR KO mice were comparable to that of WT mice. Conversely, B-VDR KO mice had higher total serum IgE levels than WT mice and, when immunized, produced significantly more OVA-IgE. In our current model, we propose that B cell-derived IL-10 is regulated by the VDR and that autocrine IL-10 is higher in WT mice, therefore inhibiting IgE production to a greater extent in WT than in VDR KO mice. The major finding of this work is that vitamin D receptor expression selectively regulates IgE levels in the serum via a direct effect of the VDR in B cells.
Probiotic preparation modulate immune system response to food proteins (IRC10P.418)
Authors: Zlotkowska Dagmara , Wasilewska Ewa 
Published: January 2023

Abstract
Probiotics are microorganisms which have been reported to have beneficial effect on human health to the different extent. Presented study uses probiotic preparation (one commercial were used) to modulate mucosal immune system response to food allergens beta-lactoglobulin and ovalbumin. Mice were orally fed with 5x107 CFU DICOFLOR® by 24 days. Then mice were orally immunized with antigen on 10, 17 and 24 day of experiment. At 35 days post-immunization, significant decreases in serum Ag-specific IgG and IgA titers were observed: 2^6 and 2^4 for β-LG immunized mice and 2^7 and 2^2 for OVA-immunized mice, respectively . Specific IgA titer in fecal extracts was slightly higher than that in groups not supplemented with DICOFLOR®. On 48th day of the experiment, DTH test was performed. No response was observed 24 and 48 hours after DTH challenge by groups supplemented with probiotics preparation. Lymphocytes from spleen, head and neck lymph nodes, Peyer’s patches and mesenteric lymph nodes were phenotyped for CD8+, CD4+and CD4+CD8+ T cells. In Peyer’s patches, mice immunized β-LG and supplemented with probiotics showed elevated numbers of CD4+CD8+ and CD8+ T cell subsets compared to β-LG group not supplemented with probiotic. Similar results were found for OVA group. Results show that probiotics has potential to modulate immune response to food allergens. Those potential could be used to prevent symptoms of allergy to food allergens.
Stromal cells from TNF-transgenic mouse inflamed lymph nodes differentially affect B cell subset maintenance and activation in vitro (IRC11P.429)
Authors: Bottaro Andrea , Kuzin Igor 
Published: January 2023

Abstract
Inflamed lymph nodes undergo major structural remodeling, associated with proliferation and functional alterations of stromal cell components. In recent years, critical roles for stromal cells have emerged in determining these architectural changes, as well as affecting the trafficking and localization of immune cell populations and the outcome of immune responses. We have shown that in the TNF-transgenic (TNF-tg) mouse model of arthritis, development of chronic inflammatory joint disease is associated with dramatic changes in the organization of draining lymph nodes and in the phenotypic and functional features of resident B cells, with characteristic expansion of a unique CD23+, CD21/35-high, CD1d-high B cell population (B-in cells). These changes in turn profoundly affect lymphatic drainage function, contributing to pathogenesis. In the studies presented here, we show that in vitro co-culture of B cells on stromal cells from TNF-tg, but not non-inflamed wild-type lymph nodes results in maintenance of the B-in phenotype, elevated surface CD23 expression, and changes in proliferative and functional responses to activation stimuli. Altogether, these results are consistent with a direct role of lymph node stroma in the B cell alterations associated with TNF-tg pathogenesis, and more broadly demonstrate that chronic inflammation-induced changes to stromal cells significantly affect the functional properties of resident B lymphocyte populations.
Expression of Helios is required for the differentiation of activated/effector T regulatory cells (IRC11P.438)
Authors: Thornton Angela , Sebastian Mathew , Korty Patricia , Lopez-Ocasio Maria , Shevach Ethan 
Published: January 2023

Abstract
Foxp3+ Regulatory T cells (Treg) suppress immune activation and play a critical role in the maintenance of self-tolerance. A subpopulation (70-80%) of Treg cells in both mouse and man express the transcription factor, Helios, but the role of Helios in Treg function is still unknown. Deletion of Helios expression in all T cells (CD4-Cre) or in early progenitors (Vav-Cre) did not result in immune dysfunction. As some studies have suggested that Helios might be expressed in activated T conventional cells, we generated mice with a conditional deletion (KO) of Helios in Treg. The selective deletion of Helios in Tregs led to a slowly progressive, systemic immune activation characterized by a Th1 profile, lymphadenopathy, splenomegaly, increased serum Ig and enhanced lymphoid follicle and germinal center formation in the absence of immune pathology. In vitro suppressive function of Helios KO Treg was normal, but KO Treg failed to differentiate into T follicular regulatory cells in vivo. While the absolute number of Treg in KO mice was enhanced, in competitive settings Helios deficient Tregs were at a striking disadvantage in the periphery. Notably, Helios KO Tregs with an activated (CD44hiCD62Llo) effector Treg phenotype were greatly decreased. Analysis of Helios KO Tregs revealed increased levels of several BH3 domain pro-apoptotic genes. These findings suggest that Helios plays a critical role in the development and/or survival of effector Tregs by repressing pro-apoptotic genes.
Regulatory T cells ameliorate acetaminophen-induced immune-mediated liver injury (IRC11P.439)
Authors: Wang Xuefu , Sun Rui , Tian Zhigang 
Published: January 2023

Abstract
The contribution of innate immune cells to acetaminophen (APAP)-induced liver injury has been extensively investigated. However, the roles of T cell populations among adaptive immune cells in APAP-induced liver injury remain to be elucidated. Herein, we found that distinct CD4+ T cell subsets but not CD8+ T cells modulated APAP-induced liver injury in mice. After APAP challenge, more CD62LlowCD44hiCD4+ T cells appeared in the liver, accompanied by increased IFN-γ. The removal of CD4+ T cells by either antibody depletion or genetic deficiency markedly compromised pro-inflammatory cytokine levels and ameliorated liver injury. Meanwhile, we also found that the frequency and absolute number of Treg cells also increased. Treg cell depletion increased hepatic CD62LlowCD44hiCD4+ T cells, augmented pro-inflammatory cytokines, and exacerbated liver injury, while adoptive transfer of Treg cells ameliorated APAP-induced liver injury. Our investigation suggests that Th1 and Treg subsets are involved in regulating APAP-induced liver injury. Thus, modulating the Th1/Treg balance may be an effective strategy to prevent and/or treat APAP-induced liver injury.
The effect of 2, 3, 7, 8-Tetrachlorodibenzo -p-Dioxin in the human 3’IgH regulatory region and immunoglobulin expression. (IRC11P.440)
Authors: alfaheeda zahra 
Published: January 2023

Abstract
The transcription of Ig heavy chain (Igh) is regulated by numerous regulatory elements including 3’Igh regulatory region (3’IghRR).Several transcription factors are involved in modulating the 3’IghRR and aryl hydrocarbon receptor (AhR). TCDD is an environmental contaminant and a ligand for AhR known to interfere with Immunoglobulins (Ig) expression in animal models. Our lab have identified in previous study the mouse 3’Ig heavy chain regulatory region (3’IgH RR) as a sensitive transcriptional target of TCDD, which inhibits mouse 3’IgH RR activation and induces AhR binding to dioxin response element (DRE) within 3’IgH RR. In human the hs1, 2 enhancer is polymorphic which has been correlated with several autoimmune diseases. We have identified TCDD-induced activation of the hu-hs1, 2 enhancer and a decrease in enhancer activity with deletion of the polymorphic region. Pervious study in our lab suggested that the hu-hs1, 2 enhancer may be a negative regulator of 3’IgH RR activity and Ig expression. The objective of this study is to study the cooperative effect of the Human 3’IgHRR Enhancers in intronic promoters and the possibility of TCDD interfering with this activity in the presence and absence of the stimulation and to study the polymorphic alleles effect in the human 3’IgHRR basal activity. We used human CL-01 B cell for this study and multiple luciferase reporter constructs driven by human Igγ intronic promoter and regulated by the human 3’IgHRR enhancers .
Regulatory T cell differentiation and Th17 T cell function are regulated by mPGES-1-dependent PGE2 (IRC11P.424)
Authors: Maseda Damian , Crofford Leslie , Johnson Elizabeth 
Published: January 2023

Abstract
Prostaglandin E2 (PGE2) is a lipid mediator that regulates inflammatory and immune responses. PGE2 has pleiotropic effects on many cell types, but it is still unclear how it affects T cell differentiation and function. The mPGES1 enzyme is part of the PGE2 biosynthetic pathway and, due to its inducible nature, it controls PGE2 concentrations during activation of the inflammatory program. Absence of mPGES1 causes shunting to alternate prostaglandins and changes the characteristics of the inflammatory response. We identified that, compared to WT, naïve T cells from mice with genetic deletion of mPGES1 express a 4-fold lower level of TFGbR1 and 8-fold lower level of the EP4 PGE receptor. Since TGF-b is a key cytokine for the generation of both regulatory and Th17 cells we addressed the effect of mPGES1 deficiency during in vitro T cell polarization. We observed that mPGES1-/- naïve CD4+ T cells generate less iTreg cells, while Th17 commitment is unaffected. However, when stimulating T cells with anti-CD3/CD28 from mice following immunization with type II collagen (CII), mPGES1-/- CD4+ T cells were unable to secrete IL-17A compared to its WT T cell counterparts. Furthermore, in-vivo antigen-specific Th17 responses from CII immunized mice were also impaired, showing a decrease in IL17A production in mPGES1-/- T cells compared to WT. Hence, endogenous production and sensing of PGE2 modulates T cell differentiation possibly by altering TGF-b responses.
ICER controls Th17 cell differentiation and autoimmune pathology (IRM15P.600)
Authors: Yoshida Nobuya , Mizui Masayuki , Otomo Kotaro , Koga Tomohiro , Bradley Sean , Comte Denis , Rosetti Florencia , Kyttaris Vasileios , Crispin Jose , Tsokos George 
Published: January 2023

Abstract
Inducible cAMP early repressor (ICER) is a splice variant of cAMP-responsive element modulator (CREM). Because ICER lacks transcriptional activation domains, ICER acts as a repressor of various genes involved in immune cell function and inhibits T cell activation by suppressing the induction of pro-inflammatory cytokines including IL-2. However, little is known about ICER involvement in IL-17 production and Th17 differentiation. Culture of murine naïve CD4 T cells under Th17 skewing conditions showed surprisingly high expression of ICER indicating and important role in these cells. T cells from animals in which the CREM gene had been deleted were cultured under these conditions, which revealed less IL-17 than T cells from CREM sufficient mice. Because ICER induction is abrogated in T cells from STAT3-deficient mice, we considered the possibility that IL6-STAT3 signaling plays a crucial role in ICER induction in Th17 T cells. We expanded our observations in vivo. We induced experimental anti-glomerular basement membrane antibody glomerulonephritis (AIGN) and experimental autoimmune encephalomyelitis model (EAE) in CREM-deficient and sufficient animals and found that CREM-deficient had diminished nephritis and encephalomyelitis. Collectively, our results suggest that the transcription factor ICER supports Th17 differentiation and promotes Th17-dependent autoimmune pathology.