bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sinomenine attenuates angiotensin II-induced autophagy via inhibition of p47-phox translocation to the membrane and influences reactive oxygen species generation in podocytes
Authors: Zhang Jingbo , Wang Weili , Cai Juan 
Published: January 2023

Abstract

Background/Aims
Sinomenine, a pure alkaloid extracted from the Chinese medicinal plant Sinomenium acutum, and sinomenine hydrochloride (SN) has been successfully used for the therapy of rheumatoid arthritis (RA) and kidney diseases. Autophagy is a cytoprotective mechanism used by podocytes and other cells to alleviate the effects of oxidative stress, and angiotensin II (Ang II) significantly promotes podocyte autophagy. However, excessive autophagy may lead to cell death and podocyte depletion. The present study evaluated the effect of SN in podocytes induced by Ang II.

Methods
Podocytes were pretreated with graded concentrations (10−8 M ~ 10−4 M) of SN and then stimulated with Ang II. The LC3B protein and the p47-phox membrane fraction were measured by Western blot. Autolysosomes were assessed by transmission electron microscopy. FACS was used to quantify the ROS produced by podocytes. The translocation of p47-phox to the membrane was investigated by immunofluorescence.

Results
The 10−8M ~ 10−4M of SN alone did not effect ROS generation or podocyte autophagy. The 10−8M and 10−6M SN attenuated Ang II-induced autophagy in podocytes. Furthermore, SN decreased the level of ROS generation in Ang II-induced podocytes via inhibition of NOX subunit p47-phox translocation to the membrane.

Conclusion
The appropriate concentration of SN attenuated Ang II-induced podocyte autophagy through ROS generation, at least in part, by regulating NOX subunit p47-phox translocation to the membrane.
Exploiting T cell co-inhibition to delay autoimmune disease recurrence
Authors: Burrack Adam L , Osum Kevin , Pauken Kristen , Fife Brian 
Published: January 2023

Abstract
Type 1 diabetes (T1D) results from T cell-mediated destruction of insulin-producing pancreatic β cells. Individuals with long-term disease are at risk of developing life-threatening complications. β cell replacement is a therapy for T1D but is limited by recurrent autoreactive T cell targeted β cell death. Thus, β cells better equipped to inhibit local T cell responses may survive longer in autoimmune recipients. Programmed-death 1 (PD-1) signaling through its ligand PD-L1 inhibits T cells, and may serve as a prominent defense in T1D. Using flow cytometric analysis, in the absence of T cells in NOD.RAG−/− mice we do not detect β cell PD-L1 expression. However, with T cells, we observed an increased proportion of β cells expressing PD-L1 in female non-obese diabetic (NOD) mice which had not developed diabetes. In addition, the majority of remaining live β cells at diabetes onset in NOD mice continue to express high levels of PD-L1. These three situations suggest that islet β cells may increase PD-L1 expression as a last line of defense to limit infiltrating T cell mediated destruction. To manipulate β cell PD-L1 expression prior to transplantation, we screened a panel of diabetes-related cytokines and found that IFN-γ enhances β cell PD-L1 expression. Unfortunately, islet transplant survival was not prolonged, which we hypothesized was due to enhanced MHC class I expression, facilitating CD8+ T cell-mediated killing. We therefore de-coupled PD-L1 from enhanced MHC I expression. Using this approach, enforced β cell PD-L1 expression delays disease recurrence. These data support our hypothesis that β cells expressing T cell co-inhibitory molecules, like PD-L1, can locally inhibit autoreactive T cells which may prevent transplant destruction.
Prophylactic and therapeutic neutralization of IL-21 bioactivity protects mice from lethality induced by human T cells in a xenogeneic mouse model of graft vs host disease
Authors: Wang Shu , Naiman Brian , Gonzalez Carlos , Herbst Ronald , Kolbeck Roland , Ettinger Rachel 
Published: January 2023

Abstract
IL-21 is a pluripotent cytokine involved in both T and B cell activation/differentiation and plays a major role in Xenogeneic graft vs. host disease (XenoGVHD) where human lymphocytes are transferred into NOD/SCID/gc mice. In order to elucidate the underlying mechanisms of IL-21 action, a neutralizing anti-human (but not anti-mouse) IL-21 mAb, was administered prior to cell transfer and the resultant effects on disease progression determined. Strikingly, IL-21 neutralization halted T cell expansion, and decreased CD4/CD8 T cell ratio resulting in complete protection to wasting and lethality. Loss of IL-21R signaling significantly inhibited several cytokines including, GM-CSF, IL-10, TNF, IFNγ, IL-17A and IL-9, while surprisingly significantly increased IL-5. Both T cell expansion and serum cytokines dramatically increased after discontinuation of anti-IL-21 treatment, resulting in weight loss and death of the mice. Anti-IL-21 was protective when administered 1 or 2 weeks post-graft, where 100% and 40% survival was found, respectively. These data demonstrate that blockade of IL-21 did not simply result in loss of T cell survival, but rather inhibited homeostatic CD4 T cell proliferation and selectively impacted cytokine production switching the response from a Th1 pathogenic to a Th2 protective response. Thus, blockade of IL-21 was not globally eradicating T cell responses, but rather shaping the immune response resulting in protection from GVHD-induced lethality.
Bim identifies tumor-reactive effector CD8+ T cells in cancer patients respond to anti-PD-1 therapy
Authors: Dong Haidong , Dronca Roxana S 
Published: January 2023

Abstract
Endogenous tumor-reactive CD8+ T cell responses to tumors are controlled by immune checkpoints. Immune checkpoint blockade strategy has demonstrated therapeutic effects in some solid cancers. However, only a portion of patients responds to this therapy and there are no biomarkers to help predict and monitor treatment outcomes. We report that Bim is a downstream signaling molecule of the PD-1 pathway that reflects the degree of PD-1 interaction with its ligand B7-H1. B7-H1 protein directly induced Bim protein up-regulation in pre-activated human CD8+ T cells via regulation of AKT activation and phosphorylation of Bim. In mouse models, genetic absence of PD-1 in T cells and B7-H1 deficiency in tumors prevented Bim up-regulation in tumor-infiltrating CD8+ T cells. Among tumor-reactive PD-1+ CD11ahigh CD8+ T cells in the peripheral blood of cancer patients, Bim expression was significantly associated with expression of PD-1 and effector cell markers (T-bet and granzyme B). The frequency of Bim+ PD-1+ CD8+ T cells significantly increased in the peripheral blood of advanced melanoma patients compared to healthy donors. In melanoma patients, high Bim levels were correlated with poor overall survival. Importantly, patients who experienced clinical benefit after anti-PD-1 therapy had higher frequency of Bim+PD-1+CD8+ T cells in the peripheral blood at baseline compared to non-responding patients; however, this cell population decreased significantly after successful anti-PD-1 therapy. Our results indicate that Bim reflects the degree of PD-1 interaction with its ligand PD-L1 in effector CD8+ T cells, and measurement of Bim levels may provide a less invasive method to monitor or predict responses to anti-PD-1 therapy.
Signaling events resulting from synchronous BCR and CD180 activation on murine B cells differ from activating CD180 or the BCR alone
Authors: Roe Kelsey , Shu Geraldine , Draves Kevin E. , Clark Edward A. 
Published: January 2023

Abstract
CD180 (RP105), expressed on dendritic cells and B cells, is a receptor closely related to TLR4. CD180 signaling does not require adaptor proteins like MyD88, instead it resembles B cell receptor (BCR) signaling, including the activation of PI3K and Akt. Recently, others and we have found αCD180 also induces the Pim-1 kinase, but much of the CD180 signaling pathway remains ill defined. We previously found that targeting antigen (Ag) to B cells via CD180, by coupling the Ag to an anti-CD180 antibody (Ag-αCD180), induces a rapid and strong Ag-specific IgG antibody (Ab) response in mice. Furthermore, IgG Ab responses were maintained in immunodeficient mice that lack mature B cells (BAFFR−/−). These studies led us to investigate the signaling events that occur within B cells upon coincident activation of the BCR and CD180. For this study we have employed B1-8hi transgenic mice, which have a BCR that binds to NP-hapten on some splenic B cells, and the K46μm17 murine B cell line, that expresses both CD180 and an NP-specific BCR. By stimulating these NP-specific B cells with αCD180, NP-isotype Ab control or an NP-αCD180 conjugate, we are defining the signaling elements specific to the activation of the BCR and CD180 together vs. single receptor ligation only. Utilizing these models, we are working to characterize the survival and proliferation signals induced, including Pim-1 and Akt, and to phenotype the B cell expansion in mice following NP-αCD180 vaccination. Data will be presented showing how CD180-BCR co-ligation differs from signaling through either receptor alone. These studies will enhance our understanding of how CD180 Ag targeting is able to induce strong humoral responses even in immunodeficient mice. (Supported by NIH grant R01AI44257).
RIG-ging the host innate immune response for vaccine adjuvant and antiviral therapy
Authors: Loo Yueh-Ming , Wilkins Courtney R. , Pattabhi Sowmya , Knoll Megan L. , Kaiser Shari , Mire Chad E. , Grigg John , Hemann Emily A , Probst Peter , Posakony Jeffrey , Ireton Renee C. , Geisbert Thomas W. , Bedard Kristin M. , Iadonato Shawn P. , Gale Michael 
Published: January 2023

Abstract
Retinoic acid inducible gene I (RIG-I) is essential for directing and priming the host immune response against many RNA viruses. A diverse small molecule library was used in a cell-based screening approach to identify drug-like compounds that could target the RIG-I signaling pathway and induce downstream IRF3 to drive innate antiviral immunity. We identified small molecule compounds that activate IRF3 to induce discrete subsets of innate immune and antiviral genes. We identified a class of hydroxyquinoline compounds that induce IRF3 activation and innate immunity of cultured cells to decrease viral RNA load and infectious virus production, thereby controlling infection by viruses of the families Flaviviridae, Filoviridae, Orthomyxoviridae, Arenaviridae and Paramyxoviridae. In addition, we identified a second class of benzo-bisthiazole compounds that directly engage and activate RIG-I to promote downstream IRF3 signaling and the expression of innate immune genes, cytokines and chemokines. Importantly, these compounds serve potent immune adjuvant functions to promote the maturation of dendritic cells to enhance antigen presentation and T cell activation. When administered to mice in combination with influenza A virus or West Nile virus vaccine, this compound conferred adjuvant activity to facilitate vaccine-mediated protection of mice from lethal virus challenge under conditions of otherwise suboptimal vaccine dose. Thus, we have identified novel small molecule RIG-I agonists that activate the innate immune response. Our studies reveal the strong potential of developing small molecule RIG-I agonists as broad-spectrum antiviral agents to control RNA virus infection and as adjuvants to potentiate vaccine immunity.
Human and rat epithelial cells inactivate extracellular influenza A virus via H2O2-dependent hypothiocyanite
Authors: Gingerich Aaron , Hanson Jarod , Dlugolenski Daniel , Pang Lan , Tripp Ralph A , Rada Balazs 
Published: January 2023

Abstract
Influenza A virus (IAV) causes ongoing epidemics characterized by high morbidity and variable mortality in several species including humans and swine. Current intervention strategies incorporate either vaccines with several strains of IAV or antiviral drugs against which the virus is developing resistance. Identifying novel anti-influenza mechanisms of the respiratory innate immune system is a potential source of influenza therapies. The first line of defense against respiratory pathogens is the respiratory epithelium. One of the innate defense mechanisms respiratory epithelial cells use to prevent infection with pathogens such as IAV is an extracellular oxidative burst at the airway surface mediated by lactoperoxidase (LPO), thiocyanate anion (SCN−), and hydrogen peroxide (H2O2) to produce antiviral hypothiocyanite ions. Cellular H2O2 is produced by two isoforms of the NADPH oxidase dual oxidase (DUOX) enzymes, DUOX1 and DUOX2. Our data has shown that differentiated, primary human or rat tracheal epithelial cells supplemented with LPO and SCN− are capable of inactivating IAV in vitro. This system has shown a 4–5 log reduction in viral titers that was eliminated by the addition of catalase (an H2O2 scavenger) demonstrating its H2O2 dependence. Additionally, we have shown that this system is capable of inactivating human-origin, swine-origin, and recombinant IAV strains. These results suggest that by modulating the DUOX system IAV infection could be prevented or at least modulated to improve viral killing thereby reducing virus shedding which could alter IAV infection dynamics within a population.
Carbohydrate specific T cell stimulation by HIV envelope glycoprotein
Authors: Sun Lina , Ozdilek Ahmet , Middleton Dustin R , Avci Fikri Y 
Published: January 2023

Abstract
HIV has been a major threat to human health and a protective AIDS vaccine has not as yet been developed. The design of the current generation of vaccines does not make use of specific scientific knowledge to maximize stimulation of critical immune cells (i.e., helper T cells) involved in producing protective antibodies. HIV-1 surface is decorated with a heavily glycosylated envelope protein called gp120, whose interaction with the CD4 molecule is the key step for the virus’s entry into CD4+ T cells. Gp120 glycans have been shown to play critical roles in Env structure, viral infectivity, antigenicity, receptor binding and HIV-1 transmission. Our goal is to identify CD4+ T cell repertoires recognizing the glycan shield of HIV-1 envelope glycoprotein, gp120. We have gained significant evidence for glycan/glycopeptide presentation on the antigen presenting cell (APC) surfaces serving as potential epitopes to induce carbohydrate-specific T cells (Tcarb) response. Additionally, we demonstrate that certain populations of CD4+ T cells differentiate between glycosylated and non-glycosylated gp120 epitopes, suggesting carbohydrate recognition by CD4+ T cells. Using the discovery of such Tcarbs and their glycan epitopes, we can design and develop knowledge-based, new-generation HIV vaccines that will elicit strong and long lasting immune response to protect from HIV. We believe our study will yield a new platform to develop a new-generation of protective HIV vaccines.
Profiling system-wide immune signaling in chronic viral infection and its response to viral clearance
Authors: Angel Cesar Joel Lopez , Furman David , Pham Edward , Fram Benjamin , Nguyen Thai , Ahmed Aijaz , Grant Philip , Maecker Holden T , Glenn Jeffrey , Davis Mark M 
Published: January 2023

Abstract
There are many billions of chronic viral infections in humans, as each of the ~7.4 billion individuals alive today each host ~10 chronic infections. The clinical significance of these infections vary widely, yet all individuals with chronic viral infections rely on persistent antiviral immunity to keep these infections in check. Over time, chronically infected individuals exhibit impaired immune function as evidenced by their poor response to vaccination and progressive T cell dysfunction – a process known as exhaustion. The impact of chronic viral infection on other immune cell types is understudied, and whether the immune landscape can be restored upon viral eradication is unknown. To address these questions, we have prospectively recruited a cohort of HCV-infected patients who underwent standard of care treatment leading to viral clearance in 12 weeks, a cohort of virologically suppressed HIV-infected patients, and an aged-matched cohort of healthy individuals. With the use of single-cell mass cytometry (CyTOF) and systems bioinformatics analyses, we comprehensively characterized the molecular pathways and functional signature underlying immune exhaustion. Here we report an immune signaling signature that differentiates individuals with chronic viral infections from uninfected individuals. While this signature is largely unaltered following viral clearance, we identified a set of functional parameters that are restorable upon successful viral eradication which represent a previously unobserved level of plasticity of the human immune system.
A role for CXCL17 in Protective Vaginal CD8+ T cell Immunity Against Genital Herpes Infection and Disease
Authors: Benmohamed Lbachir , Srivastava Ruchi , Hernández Marcela , Khan Arif A , Zlotnik Albert 
Published: January 2023

Abstract
Conventional memory CD8+ T cells include effector memory (TEM) and central memory (TCM) CD8+ T cell subsets that circulate through the blood to access lymphoid and non-lymphoid tissues in order to control peripheral infections. A distinct non-recirculating tissue-resident memory CD8+ T cell subset (CD8+ TRM) develops and is retained within peripheral non-lymphoid mucosal tissues. Chemokines play a crucial role as chemoattractants for conventional memory CD8+ T cells. However, the identity of the chemokines involved in the homing of tissue-specific mucosal resident CD8+ TRM cells remain to be fully elucidated. We screened several prominent mucosal chemokines to identify those relevant to viral infections in the vagina. This screen indicated that the mucosal chemokine CXCL17 is strongly expressed in the vagina. We used herpes simplex virus type 1 (HSV-1) to determine the role of CXCL17 in immunity in the vaginal mucosa. Following HSV-1 infection, CXCL17 is strongly induced in the vaginal mucosa, and its expression parallels an increase in the number of tissue-resident CXCR8+CD8+ TRM cells. These changes are associated with a decrease in genital herpes infection and disease. Furthermore, a CXCL17−/− mouse failed to contain genital herpes when infected with HSV-1. These results strongly suggest a pivotal role for CXCL17 in protective vaginal CD8+ T cell immunity.
Elevated CD11c and CD80 levels on peripheral B cells are associated with reduced viral set points in SIV infected infant macaques
Authors: Wood Matthew P , Lippy Adriana , Fisher Bridget , Fuller Deborah , Sodora Donald 
Published: January 2023

Abstract
With 150,000 new infections in 2015, HIV continues to infect infants at an unacceptably high rate, partly due to a lack of an effective HIV vaccine. To better understand infant immune factors contributing to oral HIV transmission and disease in infants, we monitored several markers of peripheral immune cell activation in newborn infant Rhesus macaques. Infants in this study were orally challenged weekly until infected with SIVmac251. From 4–9 weeks post infection, macaques diverged into two distinct groups designated as High Viremic (HV) (7.2×107 – 3.9×108vRNA copies/mL plasma, n=4) and Low Viremic (LV) (3.8×105 – 6.2×106vRNA copies/mL plasma,, n=4) based on viral set points. Markers of immune activation on B cells, monocytes, T cells, and dendritic cells in blood were assessed by flow cytometry through 10 weeks post-infection. At 10 weeks post infection we observed significantly higher (p=0.015) inflammatory (CD14+, CD16+) monocytes in HV macaques, which has been observed in the literature in adult macaques. Unexpectedly, we also observed striking differences between these two groups of infants regarding the activation markers CD11c+ and CD80+ on B cells. LV infant macaques had significantly higher CD11c+ (p=0.036) and CD80+ (p=0.009) B cells by 5–6 weeks post infection compared to HVs (26 fold LV vs 13.2 fold HV at 10 weeks). These findings suggest that activation of B cells may contribute to reduced viral loads in these infants, or alternatively that high levels of SIV can suppress activation of B cells. Experiments are currently underway to explore B cell/antibody mediated viral control in LV infants as these findings have implications regarding development of SIV/HIV vaccines in infants.
The role of Id3 in regulating NKT cells of the adipose tissue
Authors: Buechel Heather Marie , Piccirillo Ann , D’Cruz Louise M 
Published: January 2023

Abstract
Disorders involving obesity, such as diabetes, heart disease, and metabolic syndrome have become a predominant health issues in the United States and around the world. According to the CDC 36% of adults in the US qualify as obese, leading to a variety of negative health outcomes and billions of dollars in medical costs.
One of the hallmarks of obesity is inflammation of the adipose tissue mediated by a combination of molecules such as hormones and cytokines produced by immune cells. Natural killer T (NKT) cells are a heterogeneous population of innate lymphocytes, defined in part, by their ability to quickly release cytokines after activation. NKT cells have been implicated in the maintenance of healthy adipose tissue, and loss of NKT cells in obesity has been correlated with the increased inflammation, glucose and insulin intolerance.
As a heterogeneous population, the exact manner by which adipose resident NKT cells modulate the adipose tissue is contentious. We hope to further elucidate NKT cells’ role in adipose tissue, after activation. We also hope to define the molecules responsible for mediating NKT cells’ activity in a healthy adipose environment, with the goal of modulating these cells to improve health outcomes in the disease state.
Our first molecule of interest is the inhibitor of DNA binding protein, ID3, which has been previously implicated in NKT cell development in the thymus. Here we use both a GFP reporter and a conditionally deficient line to determine the role that Id3 plays in NKT cell activation in healthy adipose tissue.
AP-1 transcription factor reprograms T cell epigenome during activation
Authors: Barski Artem 
Published: January 2023

Abstract
Upon encountering an antigen, naïve T helper (Th) cells are activated, differentiate into several lineages and contribute to immune response. Epigenetic changes at several cytokine and other effector genes during activation has been described previously, but the mechanism behind these changes is not understood. Here, we have profiled open chromatin during human T cell activation using ATAC-Seq approach and identified the regions where epigenetic changes take place. T cell activation requires antigen signaling via T cell receptor and co-stimulation via CD28 which in turn result in nuclear translocation of NFAT and AP-1 transcription factors. For this reason, we profiled genome-wide distribution of these proteins during activation by ChIP-Seq. Our results show that 73% of activation-induced open chromatin loci contain binding sites for AP-1, whereas 39% of them bind NFAT. Over 90 % of the binding sites for NFAT were shared with AP-1. We and others have previously shown that the lack of co-stimulation results in reduction or delay of AP-1 translocation into the nucleus. In order to confirm the role of AP-1 in establishment of the open chromatin structure during activation we mapped open chromatin in the cells that were activated without co-stimulation. Our data showed that ATAC signal was reduced at the open chromatin sites bound by AP-1 in the absence of co-stimulation, suggesting that CD28 signaling and AP-1 transcription factor are involved in open chromatin formation during T cell activation.