bluuebunny/crossref_metadata_2025_split huggingface dataset:
The effects of HBV core protein on TRAIL-induced apoptosis and its molecular mechanisms (41.45)
Authors: Sun Wensheng , Du Juan , Liang Xiaohong , chen Youhai H 
Published: January 2023

Abstract
Hepatitis B virus (HBV) infection afflicts more than 300 million people worldwide .TRAIL, the TNF-related apoptosis-inducing ligand, has recently been implicated in hepatocyte death during HBV infection. The goal of our study is to determine for the first time whether HBV core protein (HBc) is involved in modulating the TRAIL apoptotic pathway. Using cck-8 and TUNEL assay we firstly reported the HBV core protein (HBc) is a potent inhibitor of TRAIL-induced apoptosis. The further study showed that HBc decreased the expression of DR5, which is one of the most important TRAIL receptors. Additionally , the dual-luciferase assay indicated HBc significantly repressed the promoter activity of the human DR5 gene.Our further results examed by the microarray analysis indicated that HBc also repressed P53 gene. Whether the HBc-p53 pathway plays a significant role in regulating DR5 expression needs to be further studied. These results indicate that HBc may prevent hepatocytes from TRAIL-induced apoptosis by blocking DR5 expression, which in turn contributes to the development of chronic hepatitis and HCC.
This work was supported by grants from the National Natural Science Foundation of China ( No.30700973, No.30772031, No.30128023 and No.30700357). Corresponding author E-mail : WSW@sdu.edu.cn. Wensheng Sun , Juan Du and Xiaohong Liang contributed equally to this work.
The role of CD11c+ hepatic dendritic cells in immue tolerance to hepatocellular carcinoma (41.48)
Authors: Zheng Hong , Avella Diego M , Schell Todd D , Jiang Yixing , Staveley-O'Carroll Kevin F 
Published: January 2023

Abstract
Immune tolerance is a major mechanism involved in the development of hepatocellular carcinoma (HCC). We have previously showed, in a mouse model of spontaneous HCC, that T cell response can be induced in mice with small HCC, but not in mice with large HCC, indicating that immune tolerance occurs when tumors grow to a certain size. The mechanisms responsible for this phenomenon were not clear. In this study, we compared freshly isolated liver dendritic cell (DC) subsets in mice with no tumor, small HCC, and large HCC. We found that total numbers of CD11c+ hepatic DC were comparable in these 3 groups. However, CD11cint B220+ subset of 'plasmacytoid' DC (pDC) significantly decreased in both small and large HCC-bearing mice. Importantly, CD11chi B220- subset of mature DC significantly increased in mice with small HCC. In contrast, CD11chi B220- subset of mature DC remains same in mice with large HCC, the CD11cint B220- subset of immature DC increased instead in these mice. The surface phenotype of each subset of DCs were examined and demonstrated higher level of CD86 and CD40 expression by CD11chi subset of mature DC comparing with CD11cint subsets. These findings indicate that modulating subset distribution of liver DC, such as inhibiting the expansion of CD11chi B220- subset of mature DC, may play a role in immune tolerance to HCC.
Homeostatic proliferation transiently rescues tolerant CD8+ T cells (41.57)
Authors: Schietinger Andrea , Fowler Carla , Teague Ryan , Tan Xiaoxia , Blattman Joseph , Greenberg Philip 
Published: January 2023

Abstract
Peripheral T cell tolerance to self antigens is required to prevent the destruction of normal tissue. Since most tumor antigens are overexpressed self antigens one critical challenge is to develop strategies that break T cell tolerance to such antigens without causing autoimmunity. We previously developed a murine T cell tolerance model in which CD8+ T cells express a transgenic TCR specific for a viral protein that is expressed as a transgene selectively in the liver. Peripheral CD8+ T cells in these mice are tolerant and do not proliferate in response to the antigen. However, strategies that "force" tolerant T cells to proliferate, for example by stimulation with IL-15, abrogated tolerance.
To understand the mechanism by which repetitive cell division rescues tolerant T cells, we studied the effect of homeostatic proliferation (HP) on tolerant T cells. Transgenic tolerant T cells were adoptively transferred into lymphopenic mice in the presence or absence of a tolerizing environment. We found that: (i) expression of self-antigen dramatically augmented HP of tolerant T cells, (ii) HP rescued tolerant T cells even in a tolerogenic environment, and (iii) rescued tolerant T cells persisted without causing autoimmunity. Intriguingly, the observed HP-dependent rescue was transient; tolerant T cells became functional during the "acute phase" of HP but subsequently reacquired their tolerant phenotype even in the absence of a tolerizing environment.
In summary, our studies suggest that HP may lead to a temporary reprieve from epigenetic repression of functional activity that is subsequently reimposed after the proliferative drive terminates.
Evaluation of dendritic cell based cancer vaccines with a huPBL-NOD/SCID/IL2r-gamma-null xenotransplant model (41.7)
Authors: Harui Airi , Kiertscher Sylvia M. , Basak Saroj K , Roth Michael D 
Published: January 2023

Abstract
Dendritic cells (DC) are antigen presenting cells that have shown great potential as cancer vaccines when tested in conventional animal models. However, relating the induction of antitumor responses in mice to the efficacy in humans has been difficult. Here-in we describe the development of a xenotransplant model to detect the capacity for a DC-based vaccine to generate human anti-tumor T cell responses in vivo. Immunodeficient NOD/SCID/IL2r-gamma-null mice allow efficient engraftment and expansion of human peripheral blood leukocytes (huPBL). Over 3 weeks, the lymphoid organs of recipient mice are reconstituted with more than 70% of the spleen cells expressing human CD45. To evaluate anti-tumor responses, mice were engrafted with huPBL, implanted with human tumor cells, and then vaccinated with autologous DC expressing tumor antigens. Spleen cells showed a normal CD4:CD8 ratio, while CD8+ cells predominated in the tumor infiltrating lymphcytes (TIL). There was a significant decrease in naïve CD45RA T cells and an increase regulatory T at the tumor site. Immunized mice showed a decrease in tumor size, an increase in tumor-specific T cells, but not complete tumor resolution. This model may allow the character of TIL and the induction of human immune responses against cancer to be more realistically evaluated in vivo. This study is funded by NIH/NIDA R01-DA03018 and by the NIH/NCI UCLA SPORE in Prostate Cancer.
Ten color flow cytometry reveals CD126 as a marker for mature SP4 and SP8 thymocytes (42.10)
Authors: Tan Chibing , Taylor Ashlee A , Van De Wiele C. Justin , Teague T. Kent 
Published: January 2023

Abstract
We have developed a 10 color flow cytometric staining protocol for the simultaneous detection and characterization of 21 thymocyte subpopulations. These subpopulations include 17 previously described subsets and 4 new subsets. Importantly, this methodology uses only commercially available monoclonal antibodies and therefore could be easily employed by any investigator with access to a standard 4 laser flow cytometer. Using this novel staining scheme we have analyzed mouse thymocytes at discreet stages of development for expression of multiple cytokine receptors. As previously described, we found IL-7Ralpha to be present on some early subsets, upregulated briefly after beta-selection, downregulated on ISP and DP cells, and then re-expressed after positive selection. IL-4Ralpha was also expressed within the early DN1 and DN2 subsets. However, its expression was downregulated prior to beta-selection (in the DN3a subset) and was re-expressed in the DP cells prior to positive selection. IL-6Ralpha (CD126) showed a dramatically different pattern of expression. It was low or negative on all thymocyte subsets with the exception of the CD24low SP4 and SP8 subsets. The CD24low cells represent the more mature SP populations and therefore CD126 expression appears to be a good marker for the detection of mature thymocytes.
Quantitative Phosphoproteomic Analysis of Cellular Signaling (42.21)
Authors: Nguyen Vinh Hai , Salomon Arthur R. 
Published: January 2023

Abstract
Although many details of T cell signaling have been described previously, a systematic, quantitative wide-scale analysis of global tyrosine phosphorylation events that are triggered by activation of the T cell receptor is lacking. Here we introduce and evaluate a new quantitative approach for phosphoproteomic analysis of signaling pathway structure. Current approaches in phosphoproteomics focus on analysis of the global phosphoproteome in a single cellular state or receptor stimulation time course experiments, often with restricted number of time points. Our approach combines genetic analysis of isogenic signaling pathway mutants with quantitative phosphoproteomic method that examines disruption of downstream phosphorylation events through a time course of receptor activation using recently developed visual pathway analysis tools. This new approach is evaluated in the context of the T cell signaling pathway and a T cell clone lacking the upstream Zap-70 tyrosine kinase and its reconstituted counterpart. In our approach, label free quantitation using normalization to copurified phosphopeptide standards is applied to assemble high density temporal data within a single cell type, either Zap-70 null or reconstituted cells, providing a list of candidate phosphorylation sites that change in abundance after T cell stimulation. Metabolic labeling of proteins using SILAC method allows for the calculation of ratios used to compare Zap-70 null and reconstituted cells across a time course of receptor stimulation, providing direct information about the placement of newly observed phosphorylation sites relative to Zap-70. Quantitative phosphoproteomic signatures indicative of downstream inhibition, downstream activation, and mutant compensation are revealed and validated with proteomic pathway visual analysis tools using established T cell signaling pathway structure as a scaffold.
Quantitative Phosphoproteomic Dissection of Immunological Signaling Pathways Applied to Mast Cell and T Cell Signaling (42.5)
Authors: Salomon Arthur R. , Nguyen Vinh , Hung Norris , Cao Lulu , Lung Jonathan , Yu Kebing 
Published: January 2023

Abstract
Although many details of immune cell signaling have been described previously, a systematic, quantitative wide-scale analysis of the global tyrosine phosphorylation events that are triggered by activation of the immune cell receptors is lacking. We have combined genetic analysis with quantitative proteomic methods and recently developed visual analysis tools to dissect the tyrosine phosphoproteome of isogenic Zap-70 tyrosine kinase null and reconstituted Jurkat T cells and to examine signaling events downstream of FceRI stimulation in mast cells. In our approach, label free quantitation using normalization to copurified phosphopeptide standards is applied to assemble high density temporal data within a single cell type, either Zap-70 null or reconstituted cells, providing a list of candidate phosphorylation sites that change in abundance after T cell stimulation. Metabolic labeling of proteins in cell culture (SILAC) enables the calculation of phosphopeptide abundance ratios between Zap-70 null and reconstituted cells across a timecourse of receptor stimulation, providing direct information about the placement of newly observed phosphorylation sites relative to Zap-70. Quantitative phosphoproteomic signatures indicative of downstream inhibition, downstream activation, and mutant compensation are revealed and validated with proteomic pathway visual analysis tools using the established T cell signaling pathway structure as a scaffold. These methods are adaptable to any cell culture signaling system in which isogenic wild type and mutant cells have been or can be derived using any available phosphopeptide enrichment strategy.
Immunorepertoire analysis by multiplex PCR amplification and high throughput sequencing (42.6)
Authors: Han Jian , Wang Chunlin , Sanders Catherine M , Yang Qunying , Wang Elijah , Gharizadeh Baback 
Published: January 2023

Abstract
An immunorepertoire is the sum of functionally diverse B and T cells in one's circulation at any given moment. Three key factors shape the personal immunorepertoire: HLA type, antigen exposure history, and immunoregulation. Two major challenges have made studying the immunorepertoire difficult: the enormous diversity of V(D)Js, and the scarcity of each VDJ in a sample. To overcome these challenges, a powerful multiplex PCR method has been developed to amplify, inclusively and unbiased, all expressed VDJs in a sample. The amplicon mixture was sequenced directly with Roche 454 platform. In one run, for example, more than 73,000 and 119,000 unique V(D)Js were obtained from B and T cells, respectively. High resolution HLA typing was also carried out with the same sequencing run. This novel technology can be used to uncover possible disease mechanisms associated with abnormal immunorepertoires, to evaluate vaccine efficiency, to identify new biomarkers, and to develop new therapeutics. An online dababase and unique analytical tools have been developed for data sharing, deposition, and mining.
Direct and indirect type-1 arginase (Arg1) induction following Mycobacterium bovis (BCG) infection (43.1)
Authors: Qualls Joseph E. , DeFreitas Ashley , Smith Amber M. , Watowich Stephanie S. , Murray Peter J. 
Published: January 2023

Abstract
M. tuberculosis infects lung macrophages (MØs) and evades immune responses by a diverse array of mechanisms. We have recently published that BCG infection triggers a MyD88-dependent Arg1 induction that suppresses NO production from infected MØs. In addition, MØ-specific Arg1 conditional knockout mice were more efficient at clearing M. tuberculosis and BCG. In the present study, we have found that while MyD88 is essential for Arg1 induction following infection, MyD88-/- MØs express robust Arg1 mRNA and protein when stimulated with supernatant from BCG-infected WT MØs. Arg1 induction stimulated with BCG supernatant correlated with enhanced activation of Stat3, but not Stat1, 4, 5, or 6. Two Stat3 activators, IL-6 and IL-10, were present in the supernatants of BCG infected WT MØs. We found the combined treatment of MØs with IL-6 and IL-10 synergistically induces Arg1 in the presence or absence of BCG infection. Consequently, we propose a model by which Arg1 is induced directly by BCG infection via MyD88 signaling, and indirectly through the autocrine/paracrine IL-6/IL-10 activation of Stat3. These data suggest that mycobacteria can condition uninfected neighboring cells to suppress NO production.
The Role of IL-10 in Mice Infected with Neurotropic Murine Coronavirus (43.14)
Authors: Trandem Kathryn A , Perlman Stanley 
Published: January 2023

Abstract
Mice infected with the neurotropic coronavirus mouse hepatitis virus (MHV), attenuated strain J2.2 develop acute and chronic demyelinating disease with histopathological similarities to multiple sclerosis (MS). The process of demyelination is immune-mediated, as RAG1-/- or RAG2-/- (mice deficient in recombination activation enzyme), which lack B and T lymphocytes, do not develop demyelination upon infection with J2.2; however, if RAG1-/- mice are reconstituted with either MHV-immune CD4 or CD8 T cells, demyelination proceeded as in the immunocompetent mouse. While the pathogenic role of virus-specific CD4 and CD8 T cells and bystander CD8 T cells in this model has been demonstrated, less is known about the immune mechanisms that are controlling this anti-viral immune response. The cytokine IL-10 has been shown to be able to suppress an excessive immune response and protect the host against immune-mediated damage in EAE, an autoimmune model of multiple sclerosis. We investigated the role of IL-10 in demyelination using IL-10-/- mice. Additionally, we characterized the time, location, and antigen-specificity of IL-10 producing cells using IL-10gfp mice. Here, we demonstrate that IL-10 has a protective role in an animal model of MS and that the majority of IL-10 producing cells are MHV-specific T-cells located at the site of infection.
Funded in part by grants from the NIH and National Multiple Sclerosis Society.
Inosine-Containing ssRNA Is a Novel Viral-Associated Immune Recognition Element (43.3)
Authors: Imani Farhad , Prater Chrissy , Thakur Sheetal 
Published: January 2023

Abstract
Inflammatory responses that are generated during viral infections are critical for protective anti-viral immune responses. The exact viral-derived structures (also known as pathogen-associated molecular patterns, PAMPs) that activate uninfected cells to induce pro-inflammatory signals are not completely characterized. To date, the viral PAMPs that are identified include dsRNA and 5' triphosphate ssRNA, which are recognized by several cellular pathways such as SR-A, PKR, RIG-I, TLR-3 and MAVS. During viral infections, the cellular enzyme ADAR-1 (adenosine deaminase acting on RNA-1) catalyzes conversion of adenines to inosines, which is a pathway for blocking viral replication. ADAR-1 activation in the infected cells leads to the presence of inosine-containing viral and cellular RNA (Ino-RNA). Here, we provide in vitro and in vivo data to affirm our hypothesis that the release of Ino-RNA from infected cells and its interaction with surrounding uninfected cells is a potent stimulus for inflammatory innate immune responses. Treatment of primary human bronchial epithelial (PHBE) cells or C57BL/J6 with in vitro synthesized Ino-RNA rapidly and potently induced several markers of innate immune response such as IFN-β, TNF-a, IL-6 and RANTES. Our data uncovers a novel strategy for uninfected host cells to detect viral infections in the surrounding tissue.
IL-10 suppresses influenza-specific Th17 responses and is detrimental during high-dose primary challenge (43.8)
Authors: McKinstry K Kai , Strutt Tara M , Hamada Hiromasa , Dutton Richard W , Swain Susan L 
Published: January 2023

Abstract
Here we study the expression and influence of IL-10 during murine influenza infection. We find that IL-10 has a minimal impact on the outcome of sub-lethal infection but that IL-10-deficient mice display dramatically increased survival compared to wild-type mice when challenged with lethal doses of virus. Administration of IL-10 receptor blocking antibodies to wild-type mice results in a similar survival advantage. Increased survival in the absence of IL-10 does not correlate with increased cellular responses, or with decreased viral titers but does correlate with improved lung function and increased expression of Th17-associated cytokines during the peak of infection.
Our studies identify highly activated, lung-resident virus-specific CD4 T cell effectors that co-produce IFNγ as the major source of IL-10 during primary influenza infection and we show that autocrine IL-10 production by responding CD4 T cells is critical in dampening influenza-specific Th17 responses.
Finally, we show that transfer of virus-specific Th17-polarized CD4 T cell effectors can protect wild-type, B cell-deficient, and T cell-deficient mice against lethal influenza challenge. We further show that Th17-polarized effectors employ novel mechanisms of protection against influenza distinct from previously established modes of protection employed by Th1-polarized CD4 T cells. Our results thus show that production of IL-10 is detrimental during high-dose primary influenza challenge, and furthermore, suggest an unexpected protective role for virus-specific Th17 CD4 T cell effectors.
Regulatory role of IFN-gamma on alveolar macrophage function during influenza infection (43.9)
Authors: Sun Keer , Metzger Dennis W 
Published: January 2023

Abstract
Secondary bacterial infection often follows pulmonary virus infection. We previously reported that pulmonary IFN-γ produced during influenza infection in mice inhibits initial bacterial clearance by alveolar macrophages. As a result, there was a 100-fold increase in bacterial burden in viral-infected WT mice 48 hr after pneumococcal challenge compared to IFN-γ-/- mice. Although IFN-γ production by CD4 or CD8 T cells alone was sufficient to inhibit bacterial clearance, TCR transgenic OT-I and OT-II mice had no detectable IFN-γ following influenza infection. Thus, virus-specific T cells are responsible for significant IFN-γ production. We found that the inhibitory effect of IFN-γ was correlated with down regulation of the class A scavenger receptor MARCO. Indeed, prior influenza infection had no effect on initial bacterial clearance in MARCO-/- mice. Inhibition of MARCO expression following influenza infection could be prevented by IFN-γ neutralization. Preliminary data suggest that IFN-γ inhibits MARCO expression by signaling through STAT1 and there is significant induction of SOCS1 and SOCS3 in alveolar macrophages during influenza infection. Ongoing work is designed to determine whether influenza-induced IFN-γ facilitates activation of a SOCS negative-feedback loop in alveolar macrophages, thereby suppressing innate protection against extracellular bacteria. (NIH grant #AI41715)
Lethal viral infection results from STAT1 but not STAT2 or IRF9 deficiency in mice and is mediated by CD4+ T-cells (44.11)
Authors: Campbell Iain L , Manders Peter , Lim Sue Ling , Terry Rachael L , Getts Meghann T , Getts Daniel R , King Nicholas JC , Hofer Markus J 
Published: January 2023

Abstract
Type I IFN (α/β) signaling is crucial in the antiviral response and is mediated by STAT1, STAT2 and IRF9 while type II IFN (γ) uses STAT1. We studied the role of STAT1, STAT2 and IRF9 in the host response to systemic infection with lymphocytic choriomeningitis virus (LCMV). In wild type (WT), STAT2 KO and IRF9 KO mice, infection was non-lethal and virus was either cleared (WT) or established persistence (STAT2 and IRF9 KO). In contrast, in STAT1 KO mice, infection was lethal with severe immune pathology with T-cells, macrophages and granulocytes and elevated expression of various cytokines, particularly IFN-γ, present in various organs. However, lethality was unaltered in LCMV-infected STAT1 KO mice that lacked the IFN-γ receptor. While clearance of LCMV in WT mice is mediated by CD8+ T-cells, depletion of these cells in LCMV-infected STAT1 KO mice did not affect lethality. However, depletion of CD4+ T-cells prevented lethality in LCMV-infected STAT1 KO mice with a considerable reduction in tissue immune pathology. In vitro stimulation assays revealed a skewing of the T-cell response to the Th17 phenotype in LCMV-infected STAT1 KO but not WT mice. These findings indicated that STAT1 has a major protective function in LCMV infection, not only being crucial for limiting viral replication and spread, but also preventing the emergence of a lethal CD4+ T-cell response possibly mediated by Th17 cells. Support : NHMRC grant 512407.