bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ku protein regulates interferon regulatory factor 1 expression in response to CpG-ODN (135.39)
Authors: Riolo Matthew , Chu Wen-Ming 
Published: January 2023

Abstract
Interferon regulatory factor 1 (IRF1) expression is induced in response to toll-like receptor 9 (TLR9) stimulation by bacterial and viral CpG-DNA. Recent studies have demonstrated that IRF1's activity is specific to promoting the expression of interleukin-12p70, but not p40, in a novel signaling pathway that is myeloid differentiation protein-88 (MyD88)-dependent yet nuclear factor kappa B (NF-κB)-independent. Further evidence suggests that DNA-dependent protein kinase catalytic subunit (DNA-PKcs), is upstream of IRF1 activation and required for its expression and subsequent production of interferon in response to CpG- oligodeoxynucleotides (ODNs) stimulation. Here we have demonstrated that regulatory protein Ku is critical for IRF1 expression in response to CpG-ODN stimulation and the subsequent production of IL12p70. In Ku70 and Ku80 knock out mice bone marrow-derived dendritic cells (BMDCs) IRF1 expression was nearly abolished in response to CpG-ODN stimulation, and IL12p70 production was reduced to one fourth that of their wild type counterparts, while IL12p40 levels showed no differences. Our results suggest a novel pathway where Ku proteins regulate DNA-PKcs activation in response to CpG stimulation critical for the expression of IRF1 and the subsequent antiviral response.
This research is supported by the GAANN Training Grant.
A Role for RIP1/RIP3-Dependent Programmed Necrosis in Innate Immune Responses Against Vaccinia Virus Infections (135.44)
Authors: Chan Francis K , Cho YoungSik , Challa Sreerupa 
Published: January 2023

Abstract
Cell death by programmed necrosis (also known as caspase-independent cell death or necroptosis) is characterized by rapid loss of plasma membrane integrity prior to the exposure of phagocytic signal. The release of endogenous "danger signals" from necrotic cells trigger inflammation and may have important physiological and pathological consequences in immune responses, inflammatory diseases and cancer growth. However, the role of programmed necrosis in immune responses and inflammation has not been adequately tested, in part because the molecular pathway that regulates programmed necrosis was ill-defined. The serine/threonine kinase RIP1 plays crucial role in programmed necrosis induced by TNF and other stimuli. We performed a RNA interference (RNAi) screen to identify downstream regulators of RIP1-dependent programmed and identified RIP3 to be another kinase critically required for TNF-induced programmed necrosis. In order to test the role of programmed necrosis in inflammation and anti-viral immune responses, we infected RIP3-deficient mice with vaccinia virus and found that the RIP3-deficient mice were highly sensitive to vaccinia virus infections. The details of these studies and the physiological implications of programmed necrosis will be discussed.
Polymorphisms associated with human Dectin-1 result in disrupted protein folding and loss of function (135.47)
Authors: Wolf Andrea J , Simmons Randi M , Goodridge Helen S , Underhill David M 
Published: January 2023

Abstract
Recognition of fungal pathogens is primarily mediated by a combination of TLRs and Dectin-1, the C-type lectin receptor for fungal β-glucans. While fungal infections in U.S. hospitals primarily arise among immune compromised patients, there has been an increased incidence in fungal infections that has been complicated by the emergence of drug resistance among fungal pathogens. Even though Dectin-1 functions as one of the primary phagocytic receptors in macrophages for fungal pathogens, the importance of Dectin-1 for controlling fungal pathogens is still unclear and may be pathogen dependent. Therefore we set out to analyze the functional consequences of single nucleotide polymophisms (SNPs) in human Dectin-1. When we mutated the full length form of the A isoform of human Dectin-1, two SNPs in the coding region resulted in a loss of phagocytic function. While both mutant forms of the protein were detectable they no longer supported phagocytosis of yeast zymosan particles in non-phagocytic cells. Analysis of alternate amino acid substitutions demonstrated an essential structural role for amino acid 223. Consequently, individuals possessing either of these mutations are potentially at a higher risk for certain fungal pathogens.
TLR7-induced protective CD8+ T cell immunity is mediated by a Type I IFN and IL-12 co-dependent pathway (135.48)
Authors: Oh Jason Zee Seug , Kedl Ross M. 
Published: January 2023

Abstract
Covalent conjugation of a TLR7 agonist to a protein antigen enhances the immunogenicity of the antigen and elicits protective CD8+ T cell responses. This study identifies divergent effects of two major cytokines induced by the TLR7 agonist-protein conjugate (conjugate), IFNα and IL-12, in leading to efficient priming of CD8+ T cells. In examining dendritic cell responses to the conjugate, we found that signaling through the Type I IFN receptor was required for the timely recruitment and accumulation of activated dendritic cells in the draining lymph nodes. Furthermore, Type I IFN receptor-deficient dendritic cells failed to cross-present appreciable levels of the antigen even though the capacity for antigen uptake was not impaired. Consequently, conjugate-induced CD8+ T cell responses were substantially reduced in Type I IFN receptor-deficient mice. In contrast, IL-12 deficiency had no effect on the capacity of dendritic cells to respond to the conjugate. Yet, IL-12 was required for the induction of CD8+ T cells. Collectively, the data suggest that there is a co-dependency of both IFNα and IL-12 during the primary response to immunization with the conjugate possibly due to distinct roles on dendritic cells and CD8+ T cells, respectively. These results point to the possibility of targeting the induction of both IFNα and IL-12 as a novel strategy in vaccine development for optimal induction of cell-mediated immunity.
NIH Grant R21: AI70038
Distinct regulatory roles of NF-kB in CpGA and CpGB-induced IFN-alpha production by plasmacytoid dendritic cells (135.50)
Authors: cui chongwei , Singh Sukhwinder , George Thaddeus , Fitzgerald-Bocarsly Patricia 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (PDC) are a major source of IFN-α production in response to the TLR9 ligand CpGA, but not CpGB. TLR9, MyD88, and IRF7 have been established to be in the major pathway of PDC IFN-α production. We found that the inhibition of NF-κB activation by SN50 or NBD conferred on CpGB the ability to induce IFN-α in PDC, while CpGA-induced IFN-α was decreased. Using the ImageStream imaging flow cytometer, we found that although CpGA-induced IRF-7 translocation was inhibited by SN50, SN50 conferred the ability of CpGB to induce IRF7 nuclear translocation. The early endosomal localization of CpG has been shown to be important in IFN-α production. We found that both CpGA and CpGB moved from early to late endosomes from 30 min to 2 hr. However, neither CpGA nor CpGB was found in either early or late endosomes in large quantities upon NF-κB inhibition. Unlike in cell lines, in PDC, TLR9 was found to be constitutively localized in the early endosomes and also recruited from ER upon stimulation with CpGA. These results suggest that NF-κB plays an inverse role in the regulation of IFN-α production by PDC in response to CpGA or CpGB stimulation. Supported by AI26806.
GSK3 β Regulates the mTOR -mediated inflammatory response in TLR-stimulated innate immune cells (135.55)
Authors: wang huizhi , Garcia Carlos A. , Martin Michael 
Published: January 2023

Abstract
The tuberous sclerosis complex-mammalian target of rapamycin (TSC-mTOR) pathway has been shown to play a critical role in regulating the inflammatory response. Here we identity the downstream signaling molecules within this pathway that are responsible for differentially controlling pro- and anti- inflammatory cytokines production by TLR-stimulated innate immune cells. Utilizing a kinase screen, we demonstrate that inhibition of mTOR by rapamycin or siRNA-mediated gene silencing results in the loss of GSK3-β (Ser 9) phosphorylation and subsequently increases its kinase activity. The ability of rapamycin to inhibit the TSC-mTOR pathway and thus augment pro-inflammatory cytokines while concurrently suppressing the anti-inflammatory cytokine IL-10 was shown to be dependent on its regulation of GSK3-beta activity. Analysis of downstream kinases of mTOR identified that p70S6K was responsible for the ability of the TSC-mTOR pathway to control GSK3 activity. mTOR inhibition in vivo induced a potent adaptive and innate inflammatory response that was abrogated by preventing rapamycin from increasing GSK3 activity. These findings identify the molecular mechanism by which the TSC-mTOR pathway regulates the host inflammatory response.
Cathepsin B activity is required for NALP3-induced caspase-1 activation, lysosomal dysfunction and cell death in macrophages. (135.81)
Authors: Bruni Antonio , Ha Soon-Duck , Franchi Luigi , Nunez Gabriel , Kim Sung O 
Published: January 2023

Abstract
The nucleotide-binding and leucine-rich repeats (NLRs) are a family of innate immune receptors sensing various pathogen- and danger-associated molecules within the cytosol. Activation of NLRs such as NALP3 induces the secretion of inflammatory cytokines such as interleukin (IL)-1ß and rapid cell death, through caspase-1 activation. To date, the mechanisms associated with NALP3 activation and cell death remain poorly defined. This study examined whether lysosomal membrane dysfunction or the lysosomal protease, cathepsin B (CTSB), are involved in cell death induced by NALP3. Wild-type (WT), NALP3-, ASC-, caspase-1- and CTSB-deficient immortalized bone marrow-derived macrophages were exposed to the NALP3 activator ATP. ATP treatment caused lysosomal dysfunction measured by loss of acridine orange, caspase-1 activation and rapid cell death in WT but not in NALP3-, ASC-, caspase-1- or CTSB-deficient macrophages. Mutant Pseudomonas aeruginosa (exotoxin U deficient) induces IPAF-dependent IL-1ß release and cell death. The mutant bacteria induced cell death and IL-1ß release in WT and CTSB-deficient macrophages but not in IPAF-, ASC- or caspase-1-deficient macrophages. Altogether, these results suggest that cathepsin B is required for NALP3-induced but not IPAF-induced caspase-1 activation and lysosomal dysfunction.
Aberrant DNA demethylation of PD-1 during chronic viral infection (136.16)
Authors: Youngblood Ben , Oestreich Kenneth J , Ha Sang-Jun J , Duraiswamy Jaikumar , Akondy Rama , Boss Jeremy M , Ahmed Rafi 
Published: January 2023

Abstract
Chronic infection results in CD8 T cell exhaustion characterized by reduced cytokine expression, cell proliferation and an inability to clear the source of antigen. Central to the development of exhausted CD8 T cells is the retained upregulation of the inhibitory receptor programmed death (PD)-1. It is now known that PD-1 upregulation in exhausted CD8 T cells is coincident with the progression of many chronic human diseases including human immunodeficiency virus (HIV), hepatitis C virus, (HCV), and Epstein Barr virus (EBV). To date, relatively little is known regarding PD-1 regulation. Using Lymphocytic Choriomeningitis virus (LCMV) infection to study anti-viral CD8 T cells in mice, we observe that DNA methylation of the PD-1 promoter is present in naïve (PD-1lo) and memory (PD-1lo) CD8 T cells, while effector cells (PD-1hi) are partially demethylated. Consistent with these observations, exhausted CD8 T cells from chronically infected mice have near complete demethylation of the PD-1 promoter. Resolution of the persistent LCMV infection results in a moderate reduction of PD-1 expression and a lack of promoter methylation. These findings provide insight into antigen induced expression of PD-1 associated with many human diseases. Moreover prolonged signals for demethylation in the antigen specific cells may result in heritable transcriptional dysregulation accounting for the deviant gene profile of functionally exhausted CD8 T cells.
A novel non-Rel subunit of NF-kB confers its regulatory specificity (136.2)
Authors: Wan Fengyi , Lenardo Michael J. 
Published: January 2023

Abstract
NF-kB regulates many important immunity, inflammation and anti-apoptosis genes. It still remains largely unknown how NF-kB activates selected gene expression. We hypothesized that other protein components could form an integral part of the NF-kB and control its action on target genes. To understand the selective and high affinity recruitment of NF-kB, we used p65 in a proteomic screen and identified ribosomal protein S3 (RPS3) as a novel non-Rel component of p65 homodimer and p65-p50 heterodimer DNA binding complexes that enhances the p65-DNA interaction. Reduction of RPS3 by RNAi impaired NF-kB function, but did not affect its nucleocytoplasmic shuttling. NF-kB activating stimuli dissociated RPS3 from its cytoplasmic berth by phosphorylation and caused a parallel translocation of RPS3 to the nucleus where it promoted NF-kB binding to gene regulatory sites. RPS3 was essential for normal expression of specific NF-kB target genes requiring p65, including key physiological events, such as IL-2 and IkBa, but not CD25 and CD69 in T cells and the induction of immunoglobulin k light chain gene expression in B cells. Thus, RPS3 is a novel non-Rel subunit of NF-kB that confers specific binding to certain kB DNAs. Our discovery sheds new light on the long-standing question of how NF-kB regulates distinct sets of genes in response to specific inducers. This research was supported by the Intramural Research Program of the NIH, NIAID.
High resolution mapping in Experimental Autoimmune Neuritis identifies Ncf-1 as a candidate gene (136.26)
Authors: Beyeen Amennai Daniel , Huberle Alexander , Öckinger Johan , Jagodic Maja , Olsson Tomas , Weissert Robert 
Published: January 2023

Abstract
We here dissect the polygenetic regulation of Experimental Autoimmune Neuritis (EAN), an animal model of Guillain-Barré Syndrome (GBS). GBS is a multifocal demyelinating disorder of the peripheral nervous system. In our work with a rat EAN model, we have crossed the EAN-susceptible DA strain with the EAN-resistant PVG for 12 generations, allowing us to investigate the relative effects of small genetic regions from the two different strains. We identified one such region, Ean6, linking to clinical disease severity. Ean6 contains the gene Ncf-1, a disease protective gene in animal models of Multiple Sclerosis and Rheumatoid Arthritis. Ncf-1 is a member of the NADPH oxidase complex and regulates T cell activation in these two models. As T lymphocytes have an essential role in EAN disease progression we sought to investigate the role of Ncf-1 in our model. We stimulated Ncf-1 through the NADPH complex with the compound phytol, resulting in a milder disease in DA rat females compared to controls. Although other mechanisms of action of phytol in inflammation can not be excluded, we suggest Ncf-1 to be a candidate gene in EAN and Guillain Barré Syndrome.
Antigen-Specific Tolerance Induction Using Genetically Modified Marginal Zone B Cells (136.35)
Authors: Zhang Ai-Hong Allan , Su Yan , Li Xin , Skupsky Jonathan , Scott David W. 
Published: January 2023

Abstract
Antigen-specific tolerance induction can be induced in mouse models of autoimmune diseases by adoptive transfer of genetically modified spleen B cells. We hypothesized that genetically modified marginal zone (MZ) B cells are more efficient in mediating tolerance induction. B cell-depletion using an IgG1 mouse anti-mouse CD20 monoclonal antibody (aCD20), which spares MZ B cells, was used to test our hypothesis. Mice were injected i.v. with IgG1 type aCD20 or isotype control IgG1. Two weeks later, spleens from control mice contained 75% follicular B cells (FO; CD19+CD23hiCD21low) and 8% MZ B cells (CD19+CD23intCD21hi). IgG1 aCD20 treatment reduced FO B cells by 98% but MZ B cells were relatively enriched (51% of the residual B cells). To test whether MZ B cells are enriched to function as tolerogenic antigen-presenting cells (APC), we used B cells from mOVA B cell transgenic mice (mOVA mice), which express membrane-bound OVA. C57Bl/6 recipient mice were adoptively transferred with 5 ×10E6 LPS-activated MZ B cells or whole spleen B cells from mOVA or B6 mice, as control. One week after B-cell transfer, the mice were challenged with OVA323-339/CFA, and T-cell proliferation response to OVA323-339 analyzed two weeks later. Adoptive transfer of LPS activated mOVA spleen B cells, but not B6 B cells, were tolerogenic and significantly decreased the T-cell response to OVA323-339 (p&lt;0.001). Importantly, transfer of LPS-activated mOVA MZ B cells further dampened the T-cell response to OVA323-339 (p&lt;0.05). Taken together, we conclude that genetically modified MZ B cells are more efficient APC in mediating tolerance induction than whole spleen B cells. (Supported by NIH RO1 AI035622 and an Innovative Grant from the Juvenile Diabetes Research Foundation)
A novel zinc finger protein represses the MHC-II transactivator (CIITA) during B cell differentiation and interacts with the polycomb complex factor EZH2 (136.36)
Authors: YOON HYESUK , Boss Jeremy M. 
Published: January 2023

Abstract
The master transcriptional regulator of MHC-II gene expression, CIITA, is silenced as mature B cells differentiate into plasma cells. The mechanism of CIITA silencing is thought to employ the plasma cell differentiation repressor BLIMP-1. However, some ex vivo differentiation systems that result in CIITA silencing suggest that additional factors may be involved. An ex vivo B cell to plasma cell differentiation system was established to identify additional CIITA silencing factor(s). cDNA microarrays were employed to differentially screen for genes expressed in plasma cells. A zinc finger protein (ZFP), which is induced early during ex vivo differentiation of primary B cells, was identified as a potential repressor of CIITA. Ectopic expression of ZFP resulted in a substantial decrease in CIITA expression. ZFP was found by chromatin IP to bind to promoter III of CIITA in a region that is also bound by BLIMP-1 in plasma cells, suggesting a role in CIITA silencing. Moreover, ZFP co-precipitated with BLIMP-1 from lysates derived from plasma cell lines. ZFP also co-precipitated with the polycomb histone methyltransferase EZH2. Intriguingly, plasma cell lines displayed the histone H3K27me3 modification associated with EZH2 at promoter III. Thus, these results suggest that ZFP may function to coordinate repression of CIITA during B cell differentiation in manner that involves the polycomb complex.
Long-range distal elements within the Ifng locus (136.41)
Authors: Collins Patrick L , Henderson Mel , Chang Shojang , Mortlock Doug , Aune Tom 
Published: January 2023

Abstract
Transgenic mice carrying an 8.6 kb Ifng transgene do not recapitulate Th1/Th2 selectivity, but mice carrying a transgene with the surrounding 190 kb of non-coding sequences do. It is unknown why Ifng requires 190 kb of regulatory sequences and space. To understand distal regulation of Ifng, we constructed transgenic mice carrying ~40 kb deletions within the 190 kb transgene. We identified two large regions which regulate Ifng expression. We then created small, 1 kb, deletions to identify specific regulatory elements. We identify two 1 kb regions at -30 kb and at -77 kb which are necessary for IFN-g transcription and repress IFN-g transcription, respectively. Our results demonstrate that multiple long-range distal regulatory elements cooperate to drive the proper temporal and quantitative expression of a single gene.
Induction of severe thyroid epithelial cell hyperplasia (TEC H/P) and fibrosis by in vitro activated splenocytes from IFN-γ-/- NOD.H-2h4 mice (137.11)
Authors: Yu Shiguang , Sharp Gordon C , Braley-Mullen Helen 
Published: January 2023

Abstract
IFN-γ-/- NOD.H-2h4 mice develop TEC H/P and fibrosis characterized by abnormal proliferation of thyrocytes with infiltration of thyroids by CD4+ and CD8+ T cells, and production of anti-mouse thyroglobulin (MTg) autoantibodies. Thyroid inflammatory T cells, especially CD8+ T cells, are the primary effectors for severe TEC H/P and fibrosis. Splenocytes from donors with severe TEC H/P cultured with MTg alone, or with MTg and IL-23 transferred similar severe TEC H/P to SCID recipients, although cells cultured with MTg and IL-23 produced more IL-17 and IL-22 than those cultured with MTg alone. The objective of this study was to determine if splenocytes from naïve IFN-γ-/- mice cultured under Th17 polarization conditions could also transfer severe TEC H/P to SCID recipients. Splenocytes from naïve IFN-γ-/- mice cultured 72 hr with MTg, TGF- β, IL-6 and IL-21 or with MTg and IL-23 transferred severe TEC H/P while the same number of cells cultured with MTg alone did not. These results indicate that cells able to transfer severe TEC H/P can be activated by culturing naïve IFN-γ -/- lymphocytes under Th17 polarizing conditions. Studies are in progress to determine the mechanisms involved in activation of effector cells that induce severe TEC H/P and fibrosis. These results should be relevant for defining potential therapeutic targets for intervention in autoimmune inflammation (VA Merit Review and NIH R56 AI074857).
Environmental causes of autoimmunity: Clues to the loss of tolerance in primary biliary cirrhosis (137.14)
Authors: Naiyanetr Phornnop , Meng Liping , Butler Jeffrey D. , Pfeiff Janice , Kenny Thomas P. , Lam Kit S. , Kurth Mark J. , Leung Patrick S.C. , Gershwin M. Eric 
Published: January 2023

Abstract
Primary biliary cirrhosis (PBC) is a human autoimmune disease that destroys small bile ducts and is characterized by antimitochondrial antibodies (AMAs) directed to the inner lipoyl domain of pyruvate dehydrogenase (PDC-E2). Our thesis is that environmental factors trigger the onset of PBC by breaking tolerance to PDC-E2. In particular, we submit that chemical xenobiotic modification of the lipoyl moiety of PDC-E2 results in creation of a neoantigen with a subsequent immune response in genetically predisposed individuals. Our previous work has been focused in two areas: the use of microarrays to identify chemical xenobiotics that react to AMAs as well as or better than native autoantigen and, second, the demonstration that a PBC-like disease can be generated in mice following immunization with our optimal xenobiotic coupled to BSA. Herein, we have focused on fine mapping of the optimal chemical compounds that mimic lipoic acid. We modified the S-S linkage ring structure of lipoic acid and conjugated an array of such chemicals on a PDC-E2 peptide backbone and then analyzed such compounds for reactivity using sera from patients with PBC and a variety of controls. Importantly, by such chemical fine mapping, we now identify 6,8-bis(acetylthio)octanoic acid as an optimal chemical structure. Finally, we note that this compound has striking structural similarity with a variety of drug metabolites.