bluuebunny/crossref_metadata_2025_split huggingface dataset:
Donepezil is emerging as a potential therapeutic treatment in improving endothelial barrier function to prevent and treat coronary artery disease in Diabetes Mellitus
Authors: Wilson Kiara D , Kasim Omodolapo , Aliyeva Leyla , Graefen Bahar , Najafova Nurana , Tauseef Mohammad , Fazal Nadeem 
Published: August 2023

Abstract
Current treatments have shown limited success in reversing DM induced vascular endothelial barrier dysfunction. Vascular endothelium forms the inner most lining of the blood vessels, regulates variety of biological processes such as angiogenesis, wound healing and host defense mechanisms. During inflammatory conditions, such as diabetes mellitus and myocardial infarction endothelial cell-cell junctions start to disrupt because of the internalization of the junctional proteins, such as vascular endothelial (VE) cadherin. This leads to the formation of minute inter-endothelial gaps, and the infiltration of protein-rich fluid and immune cells in the interstitial space. If remains unchecked, the persistent buildup of edema underlying the endothelial lining sets the stage for the serious life-threatening complications. Therefore, we hypothesized that the donepezil prevents high glucose-induced endothelial barrier dysfunction by preventing the destabilization of junctional proteins. We investigated the potential protective role of Donepezil in high glucose induced increase in human coronary artery endothelial (HCAE) permeability and wound healing process. Wound healing assay was performed by employing electric signals to both wound and monitor the healing process in endothelial monolayer. To investigate the signaling mechanism involved in the protection offered by Donepezil, we found that Donepezil prevented loss of VE-cadherin in endothelial junctions, and abrogated stress fiber formation in endothelial cell exposed to high-glucose solution. In conclusion, our study shows that Donepezil maintained endothelial barrier function and improved wound healing process in endothelial cells exposed to high glucose.
Chicago State University CTRE grant to Professor Nadeem Fazal, MD, PhD
A noncanonical NF-kB - β-catenin signaling axis in dendritic cells instructs gut inflammation
Authors: Deka Alvina , Kumar Naveen , Madan Upasna , Bhawna  , Chawla Meenakshi , Awasthi Amit , Basak Soumen 
Published: August 2023

Abstract
Dendritic cells (DCs) balance intestinal homeostasis against inflammatory insults. In particular, the noncanonical NF-kB pathway, which activates RelB:p52 heterodimers, has been implicated in DC differentiation. However, if DC-intrinsic noncanonical signaling modulates DC function in the inflamed gut remains unclear. Here we report that genetic ablation of the noncanonical signaling in DCs accumulated regulatory T cells in the lamina propria and associated lymph nodes, restraining inflammation in the colitogenic gut. Our mechanistic investigation revealed that an impaired RelB:p52 gene function, rather unexpectedly, augmented the abundance of active β-catenin via impairment of the β-catenin destruction complex. In turn, β-catenin transcriptionally upregulated Raldh2, imparting a retinoic acid gene signature and tolerogenic functions to noncanonical NF-kB-deficient DCs. Indeed, mice possessing DC-intrinsic noncanonical signaling defects exhibited an increased frequency of β-catenin+ DCs in the gut and resilience to chemical induced experimental colitis. Taken together, we chart a novel crosstalk between noncanonical NF-kB signaling and the β-catenin pathway that instructs tolerogenic DC functions and modulates inflammation in the gut.
National Institute of Immunology Core funding and BT/PR36631/BRB/10/1862/2020, Department of Biotechnology (DBT).
Deep Characterization of Human γδ T Cell Subsets Defines Lineage-Specific Traits
Authors: Mann Brendan T , Sanz Marta , Ryan Paul , Bosque Alberto , Pennington Daniel , Hackstein Holger , Soriano-Sarabia Natalia 
Published: August 2023

Abstract
Human γδ T cells represent a small fraction of T cells in peripheral blood (1–10%). They constitute a unique subset of T lymphocytes that recognize non-peptide antigens through MHC-independent presentation. Major γδ T cell subsets, Vδ1 and Vδ2 possess distinct tissue localization, antigen recognition, and effector responses. We hypothesized that Vδ1 and Vδ2 T cells possess differences at the gene, phenotypic, and functional level that distinguish them as distinct lineages. Comparisons between each subset have been limited by challenges in isolating sufficient cell numbers for characterization. Through multiparametric flow cytometry we identified phenotypic signatures that defined memory and tissue homing properties specific for each subset. The immune checkpoint marker TIGIT marked a Vδ1 T cell population enriched for higher cytotoxic marker expression. Using a stringent FACS-based isolation method, we compared cytotoxicity and gene expression profiles of highly purified human Vδ1 and Vδ2 cells. Cytotoxicity assays revealed similar killing capacity of Daudi cells possibly through dissimilar mechanisms. Microarray analysis identified shared and subset-specific genetic programs including lower expression of CD3ζ in Vδ1 T cells. Analysis of transcription factors associated with subset-specific gene profiles implicated NFAT in Vδ1 and AhR in Vδ2 T cells as key regulators of ex vivo gene expression. In confirmatory assays, using TCR-dependent and independent stimuli we show higher sensitivity to calcium-mediated NFAT activity in Vδ1 T cells while Vδ2 T cells may require additional activating stimuli. These findings will facilitate further studies into the specific contributions of Vδ1 and Vδ2 T cells to the immune system.
NIH NIAID R01 AI125097
miR-19a-3p and miR-19b-3p promote microglia activation associated with neuroinflammation following spinal cord injury
Authors: Sahebdel Faezeh , Morse Leslie , Battaglino Ricardo , Olson Julie K 
Published: August 2023

Abstract
Spinal cord injury (SCI)-induced neuropathic pain represents a significant clinical challenge that affects 10 to 30% of injured persons. Following SCI, microglia activation promotes neuroinflammation through the production of pro-inflammatory cytokines which has been suggested to play a role in neuropathic pain. MicroRNAs (miRs) can be molecular mediators of neuroinflammation. Our group has shown elevated levels of circulating miR-19a and miR-19b in persons living with chronic pain following SCI. We wanted to determine whether miR-19a and miR-19b are involved in neuroinflammation mediated by activated microglia after SCI. Microglia were incubated with inflammatory stimuli in the presence of miR-19a or miR-19b mimics and analyzed for activation. Microglia activation was enhanced in the presence of miR-19a or miR-19b mimics with increased expression of pro-inflammatory cytokines and decreased expression of suppressor of cytokine signaling, SOCS1 and SOCS3. Further, our results show enhanced signaling via NF-kB and Jak pathways. Overall, our results showed that treatment of microglia with miR-19a or miR-19b inhibited SOCS1 and SOCS3 expression and promoted expression of pro-inflammatory cytokines resulting in enhanced microglia activation. These results suggest that increased expression of miR-19a and miR-19b after SCI may be promoting neuroinflammation through activation of the microglia which contributes to neuropathic pain. These results may be useful in developing therapies targeting miRNAs to control neuroinflammation and neuropathic pain in persons living with SCI.
This study was supported by the Department of Defense (W81XWH-10-1-1043), National Institutes of Health (R01AR064793) and The National Institute on Disability, Independent Living, and Rehabilitation Research (NIDILRR, 90SI5015-01-00)
Human neutrophils are a key target of glucocorticoids
Authors: Gadkari Manasi , Tran Thai , Cao Qilin , Howe Katherine N , Randazzo Davide , Kaplan Mariana J , Franco Luis M 
Published: August 2023

Abstract
Neutrophils play central roles in immunity and inflammation. Glucocorticoids (GCs) are the cornerstone of anti-inflammatory and immunosuppressive therapy. While GCs trigger an acute rise in circulating neutrophils, little is known about the mechanisms by which GCs modulate neutrophil biology. We studied the response of 9 primary human cells to in vitro treatment with GCs. Surprisingly, neutrophils were transcriptionally the most responsive cell type, and they express proportionally the most glucocorticoid receptor (GR) transcripts. Long-read RNA-seq revealed 42 full-length GR transcript isoforms in neutrophils, twice as many as had been reported previously in all other cell types. At the protein level, neutrophils also have a unique and diverse set of GR isoforms. In contrast to the widely accepted model of GR signaling, in which GR binds GC molecules in the cytoplasm and then translocates to the nucleus, GR localization in neutrophils is almost exclusively nuclear, even in the absence of GC treatment. To validate our findings in vivo, we then performed a study of healthy volunteers, with blood collected before and serially after GC infusion. We generated a pathway-level map of the human neutrophil transcriptome at baseline and in response to GCs. The in vivo transcriptional response of human neutrophils to GCs was as strong as that observed in vitro, with a predominance of upregulation. Among the most strongly induced genes are several that are either not expressed, or expressed at very low levels, in human neutrophils at baseline. Some of the proteins encoded by these genes may have putative effects on neutrophil function, including SAMSN1 and ZBTB16. These findings indicate that human neutrophils are a key target for GC action.
Cigarette smoke-initiated oxidative stress causes NF-κB activation and lung inflammation via E3 ligase Itch-mediated Txnip degradation
Authors: Chen Chia-Ling , Wu Ying-Jung , Lin Pei-Yun , Shih Fu-Chia , Lin Chiou-Feng 
Published: August 2023

Abstract
Chronic inflammatory lung diseases including chronic obstructive pulmonary disease (COPD) are characterized by pulmonary structural changes, narrowing of the small airways, and destruction of the lung parenchyma caused by the prolonged inflammation. Sustained inflammation mediated by macrophages is deliberated the critical role in COPD pathogenesis, while inductive mechanisms of the persistent inflammation remain unclear. In this study, we revealed that thioredoxin-interacting protein (Txnip) participated in cigarette smoke-incited NF-κB activation that potentially conducted pulmonary inflammation. Cigarette smoke extract markedly inhibited Txnip expression via an MAPKs-dependent regulation, which accompanied by iNOS/NO and COX-2 induction in RAW264.7 cells. The decreased Txnip was also detected in lung tissues and macrophages obtained from smoking mice, while a higher NF-κB activation and lung inflammation had arisen simultaneously. In addition, cigarette smoke-associated oxidative stress initiated the proteasomal degradation of Txnip followed by the MAPKs-regulated NF-κB activation concurrently. The expression of E3 ligase Itch was elevated in smoking mouse lungs and in hydrogen peroxide-stimulated cells, whereas specific silencing Itch significantly attenuated Txnip degradation as well as NF-κB activation. Moreover, Txnip was distinctly suppressed in lung tissues, bronchoalveolar lavage fluid cells and peripheral blood mononuclear cells obtained from patients with COPD. Accordingly, cigarette smoke-aroused oxidative stress induced Itch-mediated Txnip degradation and further NF-κB inflammation thereby enabling the COPD pathogenesis potentially.
MOST 111-2314-B-038-120-MY3 and
MOST 111-2320-B-038-012
Connexin-43: More than just a gap junction protein. Connexin-43 controls CD4+ T cell activation and clonal expansion
Authors: Bennett Beth E , Bilokon Thomas , Kraj Piotr , Kuczma Michal 
Published: August 2023

Abstract
Activation and proliferation of antigen specific T cells depends on the interaction of multiple signaling molecules with the TCR/CD3 complex. We report that expression of connexin 43 (Cx43), a gap junction protein, is critical for the expansion of antigen-specific or self-reactive effector CD4+ T cells. While gap junction mediated intercellular communication has been the main focus of connexin research, the cytoplasmic C-terminus of Cx43 has multiple functions independent of membrane channel forming properties and may act as a docking molecule for assembling signaling complexes including the immunological synapse. We also report that antigen-specific Cx43-deficient CD4+ T cells transferred into recipient mice immunized with cognate antigenic peptide and adjuvant expanded poorly compared to wild type cells. This protected mice from experimental autoimmune encephalomyelitis (EAE). Decreased proliferation and expansion also protects lymphopenic mice repopulated with naive, Cx43 deficient CD4+ T cells, from inflammatory bowel disease. We have found that Cx43-deficient CD4+ T cells have a signaling defect affecting Erk, p38 and Akt signaling pathways. Transcriptome RNA-seq analysis comparing gene expression profiles of activated Cx43 sufficient and deficient CD4+ T cells shows that differentially expressed genes are enriched in genes involved in G protein and Ca2+ signaling, ATP metabolism, genes regulating cytoskeleton reorganization and T cell homing. In summary, our data demonstrate that Cx43 has a major impact on the generation of effector CD4+ T cells, their ability to respond to antigenic stimulation, or to precipitate autoimmune disease.
The pro-inflammatory phenotype of human macrophages is restricted by the activity of the orphan nuclear receptor Nurr1
Authors: Solís Miguel Ángel , Santana Eduardo , Muñoz José Ramón , Segovia Norma Cristina , Patiño Eduardo , Sánchez-Torres Carmen 
Published: August 2023

Abstract
The nuclear receptor related 1 protein (Nurr1) is a member of the orphan nuclear hormone receptor family whose expression has been associated with several inflammatory diseases. Macrophages are key regulators of the inflammatory processes, yet information about the role of Nurr1 in human macrophages is limited. In the present study we examined the expression and activity of Nurr1 in steady state and activated human monocyte-derived macrophages. Pro- and anti-inflammatory macrophages were derived in vitro by culture of blood monocytes with the hematopoietic cytokines GM-CSF and M-CSF, respectively. Nurr1 expression was predominant in macrophages with a pro-inflammatory phenotype. Accordingly, its expression was induced by a variety of inflammatory stimuli such as TLR-2, -3 and -4 ligands, TNF, IFN-β, and IL-4. Pro-inflammatory macrophages exposed to the Nurr1 agonist C-DIM12 decreased their production of IL-1β, IL-6, and reactive oxygen species. Conversely, Nurr1 deficient macrophages produced enhanced levels of IL-6. Mechanistically, Nurr1 agonists partially abrogated the activation of the NF-κB signaling pathway in pro-inflammatory macrophages exposed to LPS, rendering cells with a lower rate of NF-κB p65 nuclear translocation. These results suggest that Nurr1 expression is linked with the pro-inflammatory phenotype of human macrophages where it may constitute a brake to attenuate the synthesis of inflammatory mediators.
Supported by grants from Secretaria de Educación Pública-Consejo Nacional de Ciencia y Tecnología (SEP-CONACYT, #A1-S-9430)
β-catenin inhibition in tumor microenvironment macrophages promotes immunogenicity
Authors: Pundkar Chetan Y , Kang Xuejia , Babu R Jayachandra , Chen Pengyu , Suryawanshi Amol 
Published: August 2023

Abstract
Melanoma cells activate Wnt/β-catenin signaling in the tumor microenvironment (TME) to promote tumor progression and suppress dendritic cell-mediated anti-tumor immunity. However, the role of β-catenin in macrophages and its effect on anti-tumor immunity is largely undefined. In this study, using XAV939, a tankyrase inhibitor encapsulated in nanoparticles, we explored whether targeting β-catenin in macrophages can promote immunogenicity in TME. Our data show that the co-culture of macrophages with melanoma cell supernatants or melanoma cells in the presence of XAV939 nanoparticles significantly promotes the expression of CD80 and CD86, and suppresses the expression of PD-L1 and CD206 compared to macrophages treated with control nanoparticles. Similarly, XAV939 nanoparticle treatment of macrophages in co-culture conditions resulted in significantly increased TNF-α and IL-6 and reduced IL-10 production by macrophages compared to control groups. Our data suggest that targeted inhibition of β-catenin in tumor associated macrophages could overcome tolerance and promote anti-tumor immunity.
CD4 T cells from mice with alopecia areata express an effector like phenotype and can transfer disease
Authors: Connell Samuel J , Crotts Sydney , Kahl Payton , Lensing Maddison , Dix Cristina , Jabbari Ali 
Published: August 2023

Abstract
Alopecia areata (AA) is an autoimmune disease with a 2% lifetime incidence. AA presents as hair loss and is characterized histologically by the presence of CD4 and CD8 T cells surrounding the hair bulb. Although CD4 T cells comprise a majority of the cellular infiltrate, little is known about the contribution of this population to disease pathogenesis. An emerging animal model of AA involves adoptive transfer of in vitro activated and expanded bulk skin draining lymph node (SDLN) cells from AA mice, resulting in the induction of disease in recipient mice. The objective of this study was to investigate the mechanisms by which those cells transfer disease. Expanded bulk SDLN cells overwhelmingly consisted of CD3+ T cells, including both CD8 and CD4 T cells. To address whether these T cell populations could separately induce AA, we first sorted CD8 T cells from the SDLNs of AA mice and found these cells potently induced disease in recipient mice. Surprisingly, isolated CD4 T cells from AA mice also had a robust capacity to induce disease, and they did so more efficiently than CD4 T cells from unaffected (UA) mice. High dimensional flow cytometric analysis of the SDLNs indicated that an enhanced number of CD4 T cells in AA mice exhibited an effector-like phenotype. Development of AA following transfer of expanded CD4 T cells relied on the presence of endogenous CD8 T cells and host responsiveness to IFN-γ, but, in contrast, loss of IL-17 signaling appeared to have no effect. Together, these data suggest that activated CD4 T-helper type 1 effector cells contribute to the activation of CD8 T cells and the subsequent attack of the hair follicle. Further studies are needed to further define the specific mechanisms by which CD4 T cells contribute to the development of AA.
Dept of VA (I01BX004907), NIH/NIAMS (R01AR077194, K08AR069111, T32AI007485)
MOG-specific T cells skew high affinity during severe demyelinating disease after recombinant protein induction
Authors: Faust Michael A , Kolawole Elizabeth , Liu Tao , Cordero Bethanie , Zhou Zemin , Evavold Brian D 
Published: August 2023

Abstract
Demyelinating diseases can be driven by CD4 T cells specific for self-antigen derived from myelin in the central nervous system (CNS). Mouse models utilizing peptide induction protocols may restrict the breadth of T cell specificity. Here, we compare mice induced with a peptide fragment of myelin oligodendrocyte glycoprotein (MOG), recombinant extracellular domain (ECD), and recombinant full-length MOG protein, which includes the transmembrane domain. We sought to understand why mice induced with recombinant MOG suffer a more severe demyelinating disease than those induced with peptide by focusing on MOG-specific CD4 T cells in the CNS. Fluorescently-labeled peptide-MHC tetramers revealed that mice induced with ECD and protein have a greater frequency of tetramer-positive T cells than peptide induction. To more precisely define the MOG-specific T cell response, we employed the two-dimensional micropipette adhesion frequency assay (2D-MP), which, unlike tetramer, identifies low-affinity antigen-specific T cells. We confirm that 2D-MP identifies more MOG-specific T cells in all three induction protocols; however, mice induced with protein had a lower frequency of MOG-specific T cells than those induced with ECD and peptide, likely due to the epitope within the transmembrane region. 2D-MP also unveiled that T cells from mice primed with protein skew high-affinity as compared to mice primed with peptide. Ultimately, our data indicate that disease after ECD and protein induction is driven by a greater frequency of high-affinity T cells than those from mice induced with peptide. We posit that this is due to a change in antigen presentation after recombinant protein priming.
5R01NS071518-09 5R01AI110113-05
miRNAs targeting TGFβ-signaling disrupt Treg development and function in CNS autoimmunity
Authors: Rau Christina N , Severin Mary E , Lovett-Racke Amy E 
Published: August 2023

Abstract
Multiple sclerosis (MS) is a demyelinating autoimmune disease of the central nervous system. Regulatory T cells (Tregs) exhibit impaired function in MS, and microRNAs (miRNAs) are known to regulate the development and activity of these cells.
Our group recently performed a large miRNA profiling study on naïve CD4 T cells in untreated MS patients. These data revealed that large number of TGFβ-signaling pathway targeting miRNAs are upregulated in MS patients compared to healthy controls. This dysregulation limits the in vitro generation of human regulatory T cells (Tregs). Administration of TGFβ-associated miRNAs in utero or after birth resulted in reduced Treg frequencies compared to controls. miRNA-treated mice of various strains were more susceptible to CNS autoimmunity modelled by induction of experimental autoimmune encephalomyelitis (EAE) at 8–10 weeks of age. Tregs isolated from miRNA-treated mice also showed a loss of T cell receptor (TCR) repertoire diversity after induction of EAE as TCR Vβ deep sequencing revealed.
Our findings indicate that increased levels of TGFβ-associated miRNAs lead to impaired Treg development and function contributing to an increased susceptibility to CNS autoimmunity. This may provide further understanding of Treg dysfunction in MS patients.
Activated memory T cells cross-talk to naïve T cells: A mechanism for generation of autoreactive T cells
Authors: Al Moussawy Mouhamad , Abdelsamed Hossam 
Published: August 2023

Abstract
Cytotoxic CD8 T cells (CTLs) can be auto-reactive under certain environmental and genetic conditions resulting in a wide range of autoimmune diseases. Although, these cells are detected at a very low frequency in healthy subjects, the mechanism(s) responsible for their maintenance/generation is not completely understood. Previously, we demonstrated that activated/memory CD8 T cells influence the fate of autologous naïve CD8 T cells acquiring activated/memory phenotype, suggesting that these cells might be auto-reactive. Since both CD4 and CD8 T cells play a pivotal role in the pathogenesis of autoimmune diseases, we examined whether their interaction could contribute to generation of auto-reactive T cells. Here, we showed that CD8 naive cells acquired activated-memory phenotype (CCR7neg CD45RO+) in the presence of activated memory CD4 T cells. Further using Type I diabetes (T1D) tetramers, we observed ~4% of these cells are autoreactive. These findings revealed a possible mechanism where activated memory CD4 T cells are capable to communicate with CD8 naive cells and generate auto-reactive T cells, which may have implications of autoimmunity following vaccination and transplantation.