bluuebunny/crossref_metadata_2025_split huggingface dataset:
Homeodomain protein Pbx1 regulates regulatory T cell development, stability and suppressive function
Authors: Choi Seung-Chul , Park Yuk Pheel , Roach Tracoyia A , Morel Laurence 
Published: January 2023

Abstract
Pre-B cell leukemia transcription factor 1 (Pbx1) regulates multiple processes in various cell types. The Pbx1 gene is located in the Sle1a1 lupus susceptibility locus in the NZM2410 lupus-prone mouse model. The Pbx1-d dominant negative isoform lacking the DNA binding domain is more frequent in the CD4+ T cells from NZM240 mice as well as lupus patients as compared to healthy controls. In addition, the transgenic expression of Pbx1-d in CD4+ T cells reproduced the phenotypes of Sle1a1 mice, with impaired Foxp3+ Treg cell homeostasis as well as increased inflammatory CD4+ T cells. To study the role of Pbx1 in Treg cells, we generated Foxp3-specific Pbx1 knock-out mice (Foxp3-Pbx1 KO), which were bred to CD4-Pbx1-d-Tg mice to generate Pbx1-d Tg-KO mice that have Treg cells solely expressing Pbx1-d. Foxp3-Pbx1 KO mice showed reduced numbers of Treg cells with impaired suppressive activity. The numbers of natural Treg (nTrg) cells are decreased in thymus but not in spleen of Foxp3-Pbx1 KO mice, while the number of induced Treg (iTreg) cells are increased in the thymus and decreased in the periphery, suggesting that Pbx1 regulates Treg cell development and stability. The results were enhanced in Pbx1-d Tg-KO mice compared with Foxp3-Pbx1 KO mice, leading to autoimmune phenotypes, suggesting that the expression of Pbx1 functional isoform is necessary for the maintenance and function of Treg cells. RNAseq analyses with Pbx1-KO Treg cells indicated that Pbx1 regulates cell cycle, apoptosis and migration. Overall, our results suggest that Pbx1 regulates Treg cell homeostasis, stability and their suppressive functions and that Pbx1-d promotes lupus development by decreasing Treg cell development and functions.
Supported by a grant from the NIH (R01 AI045050)
Treg-specific ablation of HuR results in systemic autoimmunity associated with decreased TLR4 activation and IL-1β production
Authors: Atasoy Ulus , Fattahi Fatemeh , Ellis Jason S , Socha Sarah , Bahleda Kristin 
Published: January 2023

Abstract
The RNA-binding protein HuR (Elavl1) plays a critical role in T cell activation and function by regulating the stability and transport of its mRNA targets. To study the role of HuR in regulatory T cell (Treg) development and function, we generated the Foxp3YFP-Cre HuRfl/fl mouse in which HuR is selectively deleted in CD4+ Foxp3+ cells, a lineage with crucial roles in self-tolerance and immune homeostasis. Foxp3+ Treg-specific HuR deletion results in abnormal phenotypic features (scurfy-like mice, hair loss, stippled/banded tails, significant failure to thrive, splenomegaly and multi-organ inflammation, including lung and lymphoid organs), likely the result of reduced Treg function, a phenotype similar to human IPEX syndrome (an autoimmune disorder). Unexpectedly, our RNAseq data showed reduced levels of TLR4, IL-1β, IL-18 and NLRP3 mRNA expression in HuR deficient Tregs (Foxp3YFP-Cre HuRfl/fl). In control WT mice, in vitro LPS treatment of Tregs induced increased expression of TLR4 and IL-1β production, associated with increased nucleocytoplasmic shuttling of HuR. In contrast, the expression of TLR4 and IL-1β was significantly decreased in Tregs from Foxp3YFP-Cre HuRfl/fl mice. Homozygous Foxp3YFP-Cre HuRfl/fl mice had statistically significant reductions in Treg numbers and increased frequency of activated effector T cells. We were also able to detect IL-1β secretion from WT Tregs in vitro following LPS treatment, which was amplified by the addition of DAMP agonists (e.g., ATP or Nigericin). In contrast, HuR KO Tregs had decreases in IL-1β secretion. Together, our data show that HuR is critical for normal CD4+ Foxp3+ regulatory T cell function.
Supported by grants from NIH (R01 AI080870 and R21 AI079341).
Distinct super-enhancer elements differentially control Il2ra gene expression in a cell-type specific fashion
Authors: Spolski Rosanne , Li Peng , Chandra Vivek , Shin Boyoung , Liu Chengyu , Oh Janksuk , West Erin , Vijayanand Pandurangan , Rothenberg Ellen , Leonard Warren 
Published: January 2023

Abstract
Expression of the IL-2 receptor a-chain (IL-2Ra, also known as CD25) is tightly regulated during T cell development and in mature lymphoid populations. IL-2Ra is constitutively expressed on double negative DN2/DN3 thymocytes and regulatory T cells, but its expression is inducible on mature T and natural killer (NK) cells. The Il2ra gene is regulated in part by a super-enhancer that is extensively bound by STAT5, but whether the super-enhancer elements are differentially used to control the cell-specific expression of Il2ra in distinct cell populations is unknown. Here, we generated a comprehensive series of mutant mice harboring deletion of individual or multiple elements within this super-enhancer. Strikingly, deletions in the upstream super-enhancer region dominantly affected CD25 expression on DN2/DN3 thymocytes and Treg cells, whereas deleting an intronic region primarily decreased IL-2-induced CD25 expression on peripheral CD4+ and CD8+ T and NK cells. Thus, distinct elements control constitutive versus inducible expression of the Il2ra gene in a cell-type-specific manner rather than behaving as simple enhancers whose activity is additive/cooperative, as has been observed for the a-globin and Wap super-enhancers. Moreover, the intronic super-enhancer region was critical for chromatin looping, and deletion of this region altered super-enhancer structure throughout the locus. These results demonstrate differential functions for distinct super-enhancer elements, thereby indicating ways to manipulate CD25 expression in a cell-type specific fashion, an area of importance in autoimmune and other disease states.
TRAF3 enhances type I interferon (IFN I) receptor signaling in T cells through modulation of the phosphatase PTPN22
Authors: Hornick Emma , Wallis Alicia Michelle , Bishop Gail A 
Published: January 2023

Abstract
Type I interferons (IFN I) are among the most powerful tools host cells deploy against intracellular pathogens. Their effectiveness is due to both the rapid, directly anti-viral effects of IFN-stimulated gene products, and to the effects of IFN I on responding immune cells. IFN I signaling through its receptor, IFNAR, is tightly regulated at multiple steps in the signaling cascade. Despite the distinct phenotypic changes induced by IFN I in different immune cell types, few studies have addressed the possibility of cell type-specific differences in regulation. In this study, we show that TNFR-associated factor 3 (TRAF3) promotes robust IFNAR signaling specifically in CD4+ T cells by preventing recruitment of the negative regulatory phosphatase PTPN22 to the IFNAR complex. The balance between signals through IFNAR and other differentiating cytokine receptors strongly influences CD4+ T cell differentiation and function during infections. Our work introduces TRAF3 and PTPN22 as newly appreciated regulators key to influencing CD4+ T cell activation by IFN I.
Supported by R01 AI123107, T32 AI007260, T32 HL007344, Department of Veterans Affairs
Interferon kappa (IFNk) in keratinocytes is critical for normal wound repair and is decreased in diabetic wounds
Authors: Wolf Sonya , Audu Christopher O , Joshi Amrita D , denDekker Aaron , Melvin William J , Xing Xianying , Wasikowski Rachael , Tsoi Lam C , Gudjonsson Johann E , Kahlenberg J Michelle , Gallagher Katherine A 
Published: January 2023

Abstract
Wound repair following acute injury requires a coordinated inflammatory response. Type I interferon (IFN) signaling is important for regulating the inflammatory response post- skin injury. IFN kappa (IFNk), a type I IFN, has recently been found to drive skin inflammation in lupus and psoriasis; however, the role of IFNk in the context of normal or dysregulated wound healing is unclear. Thus, this project explores the role of IFNk in wound repair. Here, we found that IFNk expression is upregulated in keratinocytes early post-injury and is essential for normal tissue repair. Under diabetic conditions, IFNk was decreased in wound keratinocytes, and early inflammation was impaired. Further, we found that the histone methyltransferase mixed lineage leukemia protein-1(MLL1) regulates IFNk expression in diabetic wound keratinocytes via an H3K4me3 mediated mechanism. Using a series of in vivo studies with a genetically engineered mouse model(Mll1fl/fl K14cre−) and human wound tissues from patients with T2D, we demonstrate that MLL1 controls wound keratinocyte-mediated IFNk and MLL1 is decreased in T2D keratinocytes. Importantly, we find the administration of IFNk early following injury improves diabetic tissue repair. These findings have significant implications for understanding the complex role type I interferons play in keratinocytes in normal and diabetic wound healing. Additionally, they suggest IFNk may be a viable therapeutic target to improve diabetic wound repair.
Supported by MNORC P30-DK089503
Loss of Ten-Eleven Translocation 2 (TET2) reduces CCR6 expression and increases B1 B cell number in the peritoneal cavity
Authors: Dennis Emily A , Bontha Sai Vineela , Marshall Melissa A , Srikakulapu Prasad , Garmey James , Blackburn Cassidy M R , McNamara Coleen A 
Published: January 2023

Abstract
TET2 is an evolutionarily conserved dioxygenase that catalyzes the conversion of 5-methyl-cytosine to 5- hydroxymethyl-cytosine, promotes DNA demethylation and regulates transcription. TET2 has been reported to regulate germinal center formation and class switch recombination in B2 B cells, yet its role in B1 B cell subsets remains largely unexplored. To investigate the role of TET2 in B1 subsets, flow cytometry of cells from the peritoneal cavity (PEC), spleen and bone marrow of TET2−/− mice and wildtype littermate controls (n = 14/group) was performed. In addition, sort purified PEC B1a, B1b and B2 cells were analyzed for methylation status (n = 24) and RNA expression (n = 24). Results demonstrated increased B1a (p = 0.0162) and B1b (p = 0.0056) cells in TET2−/− mice in the PEC, their primary niche, but not in the spleen, while in the bone marrow only TET2−/− B1a cells were increased (p = 0.0021). RNAseq analysis of PEC B1a, B1b, and B2 cells from TET2−/− and wildtype mice revealed reduced chemokine receptor CCR6 expression in B1a (−4.879068141 fold change, 1. 8.03E-08 padj.) and B1b cells (−2.17044776 fold change, 0.34771255 padj.) from the TET2−/− mice. Further, methylation analysis showed significant hypermethylation of CCR6 in B1a and B1b TET2−/− cells compared to wildtype B1a and B1b cells. This was corroborated by gene pathway analysis of the RNAseq data which showed significant downregulation of chemotaxis and migration pathways in B1a and B1b TET2−/− cells. We conclude that loss of TET2 increases hypermethylation of CCR6 and suppresses CCR6 RNA expression in B1 cells which may impair trafficking out of the PEC to the spleen, but not necessarily the bone marrow, leading to PEC accumulation of B1 subtypes.
Supported by grants from the NIH: 1R01HL 136098-01 (McNamara, C PI), R01 HL141123 (McNamara, C PI), T32 HL007284 (Dennis, E)
The Effect of Various Stimulants on Cytokine Secretion Profiles in Freshly Isolated Peripheral Blood Mononuclear Cells (PBMC)
Authors: Lefavor Rebecca , Clarke Emer 
Published: January 2023

Abstract
Cytokines play critical roles in in cancer, in regulating autoimmune diseases and in chemically induced tissue damage repair. In response to antibody treatment or as a result of corona virus disease (COVID), a cytokine storm may occur where high levels of inflammatory cytokines are produced leading to organ damage. To understand how to regulate cytokine levels, primary cell in vitro assays may assess the effects of small molecule compounds or antibodies. We evaluated OKT3, Polyinosinic-polycytidylic acid (Poly I:C), Resiquimod, CpG oligodeoxynucleotides (CpG ODN), DynaBeads with aCD3/aCD28, Phytohemagglutinin (PHA)-M and lipoplolysaccharide (LPS) on cytokine secretion from 6 normal donors. PBMCs isolated from fresh blood using a density gradient medium were plated at 1 × 105 cells per well in the presence and absence of the test compounds for 48 hours. Supernatants were assessed for interleukin 2 (IL-2), tumor necrosis factor-α (TNFα) and interferon-γ (IFNγ). The remaining cells were assessed for viability to evaluate cytotoxicity. Flow cytometric profiles of T cell subsets were determined. The data generated from the 6 normal donors varied greatly, though the effects of the stimulants followed the same trends. The Dynabeads (CD3/28) generated the highest values of IL-2, IFNγ and TNFα in all donors. OKT3 and PHA-M, Resiquimod and LPS generated lesser amounts of the cytokines whereas Poly I:C and CpG ODN did not induce these cytokines over background levels. Only PHA-M caused a decrease in cell viability. Additionally, the release of cytokines was not associated with activation of T cells. Cytokine release assays provide a robust readout with certain stimuli and may facilitate evaluation of novel test compounds for functional activity.
Sex differences in IL-3, IL-4, and IL-7 correlation with cholesterol and triglyceride levels in African Americans with high HbA1c
Authors: Kimbro K Sean , Wamble Ezekiel , Williams Ariel 
Published: January 2023

Abstract

Background
Nearly 30 million Americans have diabetes; majority are diagnosed with type 2 diabetes (T2D). Low grade inflammation is associated with T2D which influences a network of serum cytokines. IL-3, IL-4, and IL-7 play coordinated pathogenic roles in many inflammatory diseases including T2D and non-fatty liver disease. It has been previously reported high circulating glucose influences serum cytokine levels and cholesterol metabolism. However, further studies are needed to delineate the relationship between cytokine serum levels, cholesterol, and triglycerides metabolism.

Methods
A total of 232 serum samples were collected from African Americans: 161 women and 71 men. This group consists of 95 normal HbA1c and 106 high HbA1c participants. Most participants had a BMI &gt; 30 (n=171). Cytokines were measured using multiplexing assays from Luminex X-MAP® technology.

Results
In the presence of high HbA1c, there is a trend increase in serum levels of IL-4 in both men and women. In women participants, IL-3 (R=−0.41, P=0.02) is inversely correlated to BMI, but not in men. IL-3 is positively correlated to VLDL cholesterol in women with normal HbA1c. HDL is positively correlated with IL-3 (R=0.20, P=0.04). VLDL is positively correlated with IL-4 (R=0.79, P=0.007), in men with normal HbA1c. Triglycerides were positively correlated to IL-4 (R=0.80, P=0.006) and IL-3 (R=0.56, P=0.05) in men with normal HbA1c.

Conclusion
The involvement of IL-3, IL-4, IL-7 are linked to cholesterol metabolism but are possibly regulated differently between sexes. The evidence of these correlations provides a basis of understanding that could be utilized to provide intervention to reduce the complications of T2D.
Supported by NIH NIMHD U54MD012392
IL-11 induces monocyte inflammasome activation and migration to the CNS in IL-11R+ cells in relapsing remitting multiple sclerosis (RRMS)
Authors: Seyedsadr Maryamsadat , Wang Yan , Alzoheiry Manal , Kasimoglu Ezgi , Hwang Daniel , Garifallou James , Miskin Dhanashri , Markovic-Plese Silva 
Published: January 2023

Abstract

Objective
To examine the effect of IL-11 on the transcriptome changes in the blood and cerebrospinal fluid (CSF)-derived cells from RRMS patients.

Methods
IL-11+ expressing cells in PBMC or CSF were characterized by flow cytometry (n=14 and 9 respectively). Single cell RNA sequencing (scRNAseq) was performed for IL-11R+ sorted peripheral blood-derived cells following IL-11 stimulation (1 hour) (n=3 RRMS patients) or CSF and blood samples (n=2 RRMS patients). The animal model of RRMS (RREAE) was used to test the therapeutic effect of anti-IL11 mAb.

Results
RRMS patients have an increased frequency of IL-11+ monocytes, IL-11+ and IL-11R+ CD4+ lymphocytes and IL-11R+ neutrophils in comparison to matched HCs. IL-11 expressing monocytes, CD4+ lymphocytes and neutrophils accumulate in the CSF of RRMS patients in comparison to paired blood samples. scRNAseq for IL-11R+ sorted cells following IL-11 stimulation, revealed the highest number of differentially expressed genes (DEGs) in classical monocytes with upregulation of NFKB1, NLRP3 and IL1B genes involved in the inflammasome activation. All CD4+ cell subsets had an increased expression of S100A8/9 genes involved in the inflammasome activation. ScRNAseq for IL-11R+-sorted cells from the CSF revealed that classical and intermediate monocytes upregulate multiple complement, inflammasome-induced (IL-18), and migratory genes. aIL-11 mAb in RREAE decreased clinical scores, CNS inflammatory infiltrates, demyelination, IL-11+ CD4+ cells and monocytes, and numbers of NFkBp65+, NLRP3+ and IL-1b+ monocytes within the CNS.

Conclusion
IL-11/IL-11R signaling in monocytes represents a therapeutic target in early RRMS.
The study was supported by NIH 1R01AI131238-01A1 and PA Cure SAP4100083100 grant.
The roles of IL-22 and its receptor in the regulation of inflammatory responses in the brain
Authors: Lee DaHae , Jo HyeJung , Go Cheolhyeon , Kim Yejin , Kang Jae Seung 
Published: January 2023

Abstract
Interleukin (IL)-22 is a potent mediator of inflammatory responses. The IL-22 receptor consists of the IL-22Rα and IL-10Rβ subunits. Previous studies have shown that IL-22Rα expression is restricted to non-hematopoietic cells in the skin, pancreas, intestine, liver, lung, and kidney. Although IL-22 is involved in the development of inflammatory responses, there have been no re-ports of its role in brain inflammation. Here, we used RT-PCR, western blotting, flow cytometry, immunohistochemical, and microarray analyses to examine the role of IL-22 and expression of IL-22Rα in the brain, using the microglial cell line, hippocampal neuronal cell line, and inflamed mouse brain tissue. Treatment of BV2 and HT22 cells with recombinant IL-22 increased the ex-pression levels of the pro-inflammatory cytokines IL-6 and TNF-α, as well as cyclooxygenase (COX)-2 and prostaglandin E2. We also found that the JNK and STAT3 signaling pathways play an important role in IL-22-mediated increases in inflammatory mediators. Microarray analyses revealed up-regulated expression of inflammation-related genes in IL-22-treated HT22 cells. Finally, we found that IL-22Rα is spontaneously expressed in the brain and is upregulated in in-flamed mouse brain. Overall, our results demonstrate that interaction of IL-22 with IL-22Rα plays a role in the development of inflammatory responses in the brain.
This study was funded by the NRF of Korea (No.: 2017R1A2B2010948, 2017R1A6A3A11032576, 2020R1C1C1009334)
Pancreatic Interleukin-22 Receptor Signaling is Critical in Maintaining Beta-Cell Insulin Production and is Hepatoprotective
Authors: Sajiir Haressh , Wong Kuan Yau , Mueller Alexandra , Keshvari Sahar , Wang Ran , Wiid Percival , Macdonald Graeme , Prins John , McGuckin Michael A , Hasnain Sumaira Z 
Published: January 2023

Abstract
We discovered that the cytokine interleukin-22 (IL-22) is an efficient natural inhibitor of cellular stress and exogenous IL-22 improved insulin quality in pancreatic β-cells in preclinical models of Type 2 diabetes. Importantly, IL-22 completely restored glucose tolerance, suppressed fasting hyperinsulinemia/hyperproinsulinemia, and restored insulin sensitivity in obese animals. Moreover, metabolic improvements were accompanied by significant improvements in circulating triglycerides, liver function (AST:ALT ratio) and a reduction in hepatic lipid accumulation. Whilst the IL-22 receptor, IL-22RA1 is highly expressed in the pancreas and liver, its endogenous role in these tissues remains elusive. To explore this, we generated tissue specific IL-22ra1 knockout mice lacking the receptor in pancreatic β-cells (Il-22ra1fl/fl x Ins2Cre: IL-22ra1β-cell−/−) and hepatocytes (Il-22ra1fl/fl x AlbCre: IL-22ra1Hep−/−). We assessed their metabolic phenotype including glycemic control, hepatic lipid accumulation, and hepatic markers of cellular stress, lipid, and glucose metabolism with age or with obesity. We discovered that IL-22ra1β-cell−/− animals developed hyperglycemia with age and had defective insulin secretion, which was exacerbated when on a high fat diet. Interestingly, they also had a significant increase in markers of hepatic inflammation and cellular stress. In conclusion we demonstrate, for the first time, that local endogenous IL-22 secretion in pancreatic β-cells maintains insulin biosynthesis. Moreover, our work highlights the importance of the pancreatic-β-cell-liver axis as the absence of pancreatic IL-22ra1 signaling leads to a marked increase in liver inflammation and cellular stress.
Perturbations of Marrow Stromal Cell function during Acute Inflammation
Authors: Matkins Victoria , Camacho Virginia , Hoang Ashley , Patel Sweta , Welner Robert S 
Published: January 2023

Abstract
Mesenchymal stromal cells (MSCs) reside in a complex network of blood cells and can differentiate into chondrocytes, osteoblasts, and adipocytes. MSCs differentiation process is tightly regulated through intrinsic and extrinsic signals within the marrow microenvironment, leading to transcriptional programming of balanced lineage output. Inflammatory insults to this system can occur through the upregulation of activating cytokines. Pro-inflammatory cytokines’ impact on the hematopoietic lineage is well-defined, but similar impacts to stromal cells was largely undefined. To delineate these changes, we studied two well-characterized forms of inflammation: polyinosinic-polycytidylic acid (Poly(I:C)), that activates IFNα, and Lipopolysaccharide (LPS), that promotes IFNγ. Prior studies of inflammatory conditions showed bone loss; therefore, we hypothesized that there would be an increase in the MSC pool to compensate for loss of bone maturation. Using flow cytometry and lineage-tracing models of stroma, we characterize the non-hematopoietic cells’ changes following inflammatory perturbation. Our data show an increase in MSCs with decreases in osteoblasts and adipocytes following stimulation. Furthermore, we used reverse-phase protein and cytokine arrays to map the signaling mechanisms and activated pathways. We found an increase in the NFκB pathway with an accompanying increase IL-1β. Targeting of the IL-1 pathway did not phenotypically reverse the stromal perturbations but corrected the MSCs functionally. Understanding these alterations within the bone marrow microenvironment during inflammation will allow for therapeutic interventions to mediate hematopoietic diseases such as sepsis or leukemia.
Supported by NIH R01 HL150078