bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of interferon-λ1 on the secretion of IP-10 by hepatitis C virus-infected cells and the migration of natural killer cells
Authors: Yoon Joo Chun , Lee Woojeong 
Published: January 2023

Abstract
Hepatitis C virus (HCV) is an important human pathogen because it causes chronic hepatitis, liver cirrhosis, and hepatoma. Interferon (IFN)-λ1, which inhibits viral replication, and natural killer (NK) cells, which kill virus-infected cells, would be important in the innate immunity to HCV infection. Although it cannot directly stimulate NK cells because of the absence of its receptors on NK cells, IFN-λ1 is able to bind the receptors on HCV-infected hepatocytes. This study investigated, therefore, whether the stimulation of HCV-infected hepatocytes by IFN-λ1 treatment modulated NK cell function. Cell-culture-generated HCV virions, hepatoma cell lines, such as Huh7.5 and HCV-core replicon Huh7 cells, and primary human NK cells were employed. IFN-λ1 responses and NK cell function were measured by qPCR, ELISA, western blot, migration assay, and flow cytometry. IFN-λ1 effectively reduced HCV replication and increased the expression of interferon-stimulated genes (ISGs), such as MX1 and IFIT1, in HCV-infected cells and replicon cells. In addition, IFN-λ1 enhanced the transcription of chemokines such as IP-10 and IL-8. In particular, IP-10 secretion by HCV-infected Huh7.5 cells and HCV-core replicon Huh7 cells was significantly increased after IFN-λ1 treatment. Although the concentration of IP-10 secreted by HCV-infected Huh7.5 cells in vitro was insufficient to elicit considerable migration of NK cells, recombinant IP-10 could induce migration of NK cells ex vivo. In conclusion, IFN-λ1 enhanced IP-10 secretion by HCV-infected cells and IP-10 could modulate NK cell migration. These results suggested that IFN-λ1 would indirectly regulate the chemotaxis of NK cells through augmenting IP-10 secretion by HCV-infected cells.
Interleukin-36 cytokines regulate antiviral immunity in the skin and keratinocytes
Authors: Jensen Liselotte E. , Wang Peng 
Published: January 2023

Abstract
Pathogens develop immune evasion strategies that delay host immune responses. How the host has adapted counter strategies is often less clear. Herpes simplex virus (HSV) blocks inflammatory signaling from infected cells by preventing extracellular release of IL-1β. Previously, we reported that HSV infected keratinocytes retain the ability to signal danger to the surrounding tissue through secretion of IL-1α. This inflammatory signal appeared essential in vivo for early recruitment of immune cells to infected sites. Furthermore, mice deficient of the common receptor for IL-1α and IL-1β, IL-1R1, were more susceptible to lethal outcome of HSV infection. Hence, IL-1α may have evolved to counteract the HSV immune evasion mechanism targeting IL-1β. Here, the IL-1 related IL-36α, IL-36β and IL-36γ are studied. Like the IL-1 cytokines, the IL-36 proteins lack signal peptides for conventional protein secretion. The IL-36α mRNA was the principal IL-36 induced in keratinocytes by HSV-1 in vitro. Surprisingly, using the flank skin HSV-1 infection model, IL-36α knockout (KO) mice developed disease indistinguishable from that observed in wild type mice. In contrast, IL-36β KO mice progressed to more severe disease, whereas IL-36γ KO mice only developed modestly larger skin lesions. This could suggest that HSV-1 promotes retention of IL-36α in a manner resembling that restricting IL-1β activity. IL-36β on the other hand may achieve the highest degree of activity through a yet to be determined mechanism. Viral loads were elevated in skin from IL-36β KO mice. Furthermore, pre-treatment of human keratinocytes delayed progression of HSV-1 infections. These observations suggest that IL-36β regulates innate antiviral mechanisms in keratinocytes.
Spontaneous translocation of a human enterococcal gut pathobiont drives systemic autoimmunity
Authors: Vieira Silvio Manfredo , Hiltensperger Michael , Kumar Varun , Zegarra-Ruiz Daniel , Dehner Carina Anja , Barbieri Andrea , Jain Dhanpat , Goodman Andrew , Kriegel Martin A. 
Published: January 2023

Abstract
Host-microbiota interactions in the pathogenesis of autoimmunity remain poorly understood. Here, we show that a gut commensal, Enterococcus gallinarum, reaches lymphocytes beyond the gut barrier in the mesenteric lymph node, liver and spleen of lupus-prone (NZWxBXSB)F1 mice. Oral vancomycin treatment suppressed growth of E. gallinarum in tissues of these mice, prevented organ manifestations and mortality by lowering pathogenic autoantibodies, Th17 and Tfh cells. Hepatocyte-commensal cocultures revealed induction of the lupus signature cytokine IFN-α as well as the AhR pathway (AhR, Cyp1a1, Cyp1a2) that could be linked to Th17 induction in vivo. E. gallinarum monocolonization in germ-free C57BL/6 animals allowed for translocation to internal organs and induction of autoantibodies as well as increase of lamina propria plasmacytoid dendritic cells (pDCs) and systemic Th17 cells. Additionally, RNA-Seq of short-term ileum E. gallinarum-monocolonized mice demonstrated induction of molecules related to barrier function (occludin, claudins, Plvap, Axin2), the mucus layer (mucin, Fut2), antimicrobial defence (Reg3b, Defa2), and inflammation and pDC induction (AhR/Cyp1a1, Enpp3). Species-specific PCR of sterilely obtained liver tissue from autoimmune hepatitis and lupus patients revealed also E. gallinarum, suggesting similar processes in humans. Accordingly, human hepatocyte-commensal coculture demonstrated production of the same autoimmune-promoting factors as with murine hepatocytes. Collectively, these data indicate that a human pathobiont translocates spontaneously to promote autoimmunity in genetically predisposed hosts, broadening our understanding of autoimmune host-microbiota interactions.
Gut commensal-dependent production of autoantibodies against the primordial lupus autoantigen Ro60 in the absence of mammalian Ro60.
Authors: Datta Rahul , Dehner Carina Anja , Ruff William Edwards , Greiling Teri , Kriegel Martin A. 
Published: January 2023

Abstract

Introduction
The RNA-binding protein Ro60 is a primordial autoantigen in systemic lupus erythematosus (SLE), which is highly conserved across multiple species. We have previously linked anti-human Ro60 T cell and antibody responses to human commensals carrying Ro60 orthologs. Here, we tested in vivo if these autoantibodies can arise due to the murine gut microbiota in the absence of host Ro60 protein.

Methods
Lupus-prone TLR7 tg B6 mice were crossed to Ro60 KO B6 mice and followed longitudinally for Ro60 IgG ELISAs. Littermates were treated with broad-spectrum antibiotics (vancomycin, neomycin, ampicillin, metronidazole) to determine if depletion of the gut microbiota affects anti-Ro60 autoantibody induction and disease manifestations. High-throughput 16S rRNA sequencing of the V4 region was performed on the MiSeq platform.

Results
TLR7 tg mice produced persistently anti-Ro60 autoantibodies despite crossing to Ro60 KO mice. Oral administration of antibiotics significantly lowered anti-Ro60 autoantibodies (n=6 each, p=0.007). 16S sequencing of antibiotics-fed mice demonstrated outgrowth of Streptophyta but suppression of taxa that may contain commensal Ro60 orthologs (Clostridiaceae, Bacteroides).

Conclusions
Mice genetically prone to SLE-like disease can mount autoimmune responses against Ro60 even in the absence of the autoantigen. These findings suggest an antigenic trigger within the microbiota, which is supported by interventional studies with antibiotics and 16S sequencing. In summary, these studies suggest a mechanism for the development of lupus which extends beyond the known traditional risk factors.
OX40 signaling Induces Canonical Antigen Presentation-independent Proliferation of Human and Murine Thymic Regulatory T-cells
Authors: Kumar Prabhakaran , Marinelarena Alejandra , Bhattacharya Palash , Epstein Alan L. , Prabhakar Bellur S. 
Published: January 2023

Abstract
Regulatory T-cells (Tregs) play a pivotal role in immune-tolerance, and loss of their function can lead to autoimmunity. Natural Tregs generated in the thymus contribute substantially to the Treg pool in the periphery where they suppress self-reactive Teff responses. Recently, we showed that OX40L (TNFSF4) can drive selective proliferation of peripheral Tregs independent of canonical antigen presentation (CAP-independent) in the presence of low dose IL-2. Therefore, we reasoned that OX40 signaling might be integral to the TCR-independent phase of murine and human thymic Treg (tTreg) development. Thymic Treg development is a two-step process: Strong T-cell receptor (TCR) signals combined with co-signals from the TNF receptor Super Family (TNFRSF) members facilitate tTreg precursor selection, followed by a TCR-independent phase of tTreg development when their maturation is driven by IL-2. Therefore, we investigated whether OX40 signaling can also play a critical role in the TCR-independent phase of tTreg development. OX40−/− mice had significantly reduced numbers of CD25−Foxp3low tTreg precursors and CD25+Foxp3+ matured tTregs, while OX40L treatment of WT mice induced significant proliferation of these cell subsets. Relative to tTeffs cells, OX40 was expressed at higher levels in both murine and human tTreg precursors and matured tTregs. In ex vivo cultures, OX40L increased tTreg maturation and induced CAP-independent proliferation of both murine and human tTregs which was mediated through the activation of AKT-mTOR signaling. These novel findings show an evolutionarily conserved role for OX40 signaling in tTreg development.
Ocular autoimmunity develops despite concurrent absence of Th1 and Th17 signature cytokines
Authors: Bing So Jin , Silver Phyllis B. , Jittayasothorn Yingyos , Horai Reiko , Caspi Rachel R. 
Published: January 2023

Abstract
Th1 and Th17 cells are each a standalone effector phenotype, and each can elicit ocular autoimmunity independently of the other. In this study, we used mice deficient in IL-17A, or IFN-gamma (GKO), or both (DKO) to dissect the role of Th1 and Th17 cytokines in experimental autoimmune uveitis (EAU). EAU was induced by active immunization with the retinal autoantigen IRBP. Previous studies showed that EAU in GKO mice was exacerbated, whereas it was attenuated in IL-17A KO mice. Surprisingly, DKO mice were fully susceptible to EAU with scores similar to wild type (WT) controls, and displayed eosinophil-dominant infiltration along with enhancement of IL-17F, Th2, and proinflammatory cytokines. To test which cytokines contributed to pathology, EAU-challenged DKO mice were treated with blocking antibodies to IL-17F, TNF-a, GM-CSF or IL-4Ra (a shared subunit of receptor for IL-4 and IL-13). Blockade of IL-17F or TNF-a in DKO mice did not affect the severity of EAU. Treatment with an anti-GM-CSF antibody during both induction and expression stages significantly suppressed disease severity in DKO, but not in WT mice. By contrast, treatment of DKO mice with an anti-IL-4Ra antibody worsened EAU. Importantly, a single intravitreal injection of an anti-GM-CSF antibody after the disease onset reduced EAU severity in both WT and DKO mice. These results suggest that, in the absence of both IFN-gamma and IL-17A, (a) GM-CSF plays a major and nonredundant role as a pathogenic cytokine, (b) the Th2 cytokines IL-4 and/or IL-13 are protective, and (c) IL-17F and TNF-a may be dispensable. Our results highlight that ‘classical’ Th1 and Th17 cells are not the only pathogenic effectors, and that other inflammatory cytokine(s) can contribute to uveitis.
Islet-infiltrating effector and regulatory T cells specific for a single epitope possess distinct, but overlapping TCR repertoires.
Authors: Jing Yi , Kong Yuelin , Bettini Matthew L. , Bettini Maria 
Published: January 2023

Abstract
Regulatory T cells (Tregs) have been shown to play a critical protective role in type 1 diabetes. However, TCR specificity of islet-infiltrating Tregs and its influence on T cell function during autoimmune diabetes is largely unknown. It has been shown that Tregs have a higher affinity for self and possess a unique TCR repertoire. Accordingly, we observed increased TCR signaling in islet-infiltrating insulin-tetramer labeled Tregs compared to effector T cells (Teffs), as measured by the Nur77-GFP reporter of TCR activation. In order to obtain the profile of Treg and Teff TCR repertoires specific for a single epitope, we isolated and sequenced insulin tetramer binding Teffs and Tregs from islets of NOD mice expressing a fixed alpha chain of an insulin specific TCR. Surprisingly, TCR sequence analysis showed relatively high similarity (Morisita-Horn index: 0.27) between Treg and Teff TCR repertoires, where a substantial portion (46.4%) of Tregs expressed shared TCRs. Collectively, our data suggest that Treg and Teff specific for a single epitope can express the same TCRs, which they exploit to exert different effector or suppressive functions.
Cell-specific metabolic models reveal novel metabolic regulators of Th17 pathogenicity: from single-cell RNA-Seq to actionable metabolic targets
Authors: Wagner Allon , Wang Chao , DeTomaso David , Koul Arman , Regev Aviv , Kuchroo Vijay K. , Yosef Nir 
Published: January 2023

Abstract
Cellular metabolism is a powerful regulator of immune response. In order to unbiasedly search for novel metabolic regulators of Th17 development and function, we have developed COMPASS, a computational algorithm to characterize the metabolic landscape of single cells based on single-cell RNA-Seq profiles and flux balance analysis. We used COMPASS to characterize the metabolic heterogeneity in Th17 cells, whose pathogenic state triggers auto-immunity, yet whose non-pathogenic form promotes tissue homeostasis and barrier functions. COMPASS recovered known metabolic switches and predicted that the polyamine pathway should be a novel, powerful regulator of Th17 pathogenicity. We validated the pathway’s effect through an array of transcriptome, LC/MS metabolome, and functional assays. Deletion of polyamine enzymes in T cells resulted in altered metabolic space, T cell functions and, most importantly, aggravated symptoms in EAE, a murine model of multiple sclerosis. While our study is concerned with Th17 cells, COMPASS is generally applicable, and can be used to unbiasedly characterize the metabolic states of any cell population based on its single-cell transcriptome profiles. Furthermore, COMPASS predicts metabolic switches between cell states that present testable, mechanistic hypotheses.
Diet-induced obesity impairs induction of Experimental Autoimmune Encephalomyelitis and is restored by PD-1 blockade
Authors: Le Catherine , Khuat Lam T. , Dunai Cordelia , Soulika Athena , Murphy William J. 
Published: January 2023

Abstract
Obesity has marked effects on immune function and is associated with increased infection susceptibility and decreased adaptive immune response. We examined the effects of obesity on adaptive autoimmune responses using a diet-induced obesity mouse model. The pathogenesis of experimental autoimmune encephalomyelitis (EAE), a model of autoimmune demyelinating disease of CNS, is mediated by autoreactive Th1 and Th17 CD4 T cell subsets. 8–9-month-old C57Bl/6 male mice were fed 60% fat or 10% fat control diet starting at 6–8 weeks old and were immunized with 300 μg of rodent myelin oligodendrocyte glycoprotein (MOG)35–55 peptide in complete Freund’s adjuvant (CFA) containing 5 mg/ml heat-killed M. tuberculosis. Control diet mice exhibited onset of severe clinical symptoms of hind limb paralysis similarly to young wild-type controls, while diet-induced obese mice experienced a significantly delayed onset. Clinical scores correlated with greater number of MOG-specific tetramer-positive CD4 T cells and number of IFN-gamma and IL-17a-expressing CD4 T cells in the CNS of control mice compared to obese mice. Decreased functional cytokine expression and proliferation called into question the exhaustive nature of T cells in obese mice through the upregulation of the PD-1/PD-L1 pathway, a known regulator of EAE. We treated EAE-induced mice on control and high-fat diet with anti-PD-1 blockade (29F.1A12) and restored EAE induction in both clinical score and number of pathologic CD4 T cells in the CNS of obese mice. Our findings may indicate an impairment of the adaptive immune system through the PD-1/PD-L1 pathway in obesity, possibly indicating an augmentation of the checkpoint pathway in settings of systemic inflammation.
Identifying novel regulators of T cell CNS infiltration in Experimental Autoimmune Encephalitis
Authors: Salzler Harmony R. , Carr Tiffany M. , Wheaton Joshua M. , Ciofani Maria 
Published: January 2023

Abstract
T helper 17 (Th17) lymphocytes promote immunity in many contexts, including infection clearance, mucosal barrier homeostasis, and autoimmunity. IL-17A producing Th17 cells differentiate from naïve precursors upon antigen encounter with specific cytokines. One important mediator of Th17 cell differentiation downstream of cytokine signaling, is the AP-1 family transcription factor, JunB. JunB is a master regulator of Th17 cell fate, both promoting expression of Th17 lineage-specific genes and suppressing those of alternative effector fates. Studies in mice with JunB deficient CD4 T cells reveal that JunB is a key arbiter of pathogenesis in the experimental autoimmune encephalitis (EAE) model of autoimmunity. In addition to displaying an increased proportion of alternative T helper cell fates, JunB CD4 T cell knockout (KO) mice are completely rescued from EAE symptoms. CD4 T cells fail to infiltrate the central nervous system (CNS) in these animals, despite retaining a significant population of IL-17A producing cells in other niches. Thus, JunB-deficient mice represent a compelling model for investigating the molecular determinants of encephalitogenic T cell infiltration into the CNS. Additionally, in vitro RNAseq experiments comparing JunB KO and WT cells differentiated under Th17 polarizing conditions, suggest several promising migration related JunB target genes which are currently under investigation. Discovery of one or more JunB targets mediating this migration defect is of great potential interest in developing targeted therapies for Th17 mediated autoimmunity.
Autoantibody-inducing CD4 T (aiCD4 T) cell that induces systemic lupus erythematosus (SLE) includes not only follicular helper T cell but also IL-21-producing CXCR5−ICOShiPD-1hi cell
Authors: Tsumiyama Ken , Shiozawa Shunichi 
Published: January 2023

Abstract

Objective
We found that repeated immunization with antigen induces systemic lupus erythematosus (SLE) in mice, and we have then proposed a novel ‘self-organized criticality theory’ that autoimmunity takes place when host’s immune system is overstimulated by repeated exposure to antigen to levels that surpass the immune system’s stability limit. A novel T cell type which we termed autoantibody-inducing CD4 T (aiCD4 T) cell was newly generated via TCR revision at periphery. They not only stimulated B cells to generate varieties of autoantibodies but also helped differentiation of CD8 T cell into CTL to induce tissue injuries. We also showed that the aiCD4 T cell belongs to CD45RBlo122lo CD4 T cell subpopulation. Here we further dissected the phenotype of aiCD4 T cell.

Methods
BALB/c mice were repeatedly immunized with OVA and SLE was induced. Cell surface marker of CD45RBlo CD4 T cell was detected using flow cytomery. These CD4 T cells were stimulated with anti-CD3 and CD28 antibodies in vitro, and cytokine in culture supernatant was detected using ELISA.

Results
After repeated immunization with OVA, CD4+CXCR5+ICOShiPD-1hi follicular helper T (Tfh) cell was increased, and these Tfh cells were CD45RBlo. Further, CXCR5−ICOShiPD-1hi cell was also increased in CD45RBlo CD4 T cell of OVA-immunized mice. We found that CXCR5−ICOShiPD-1hi CD4 T cell highly produced IL-21 rather than Tfh cell after repeated immunization with OVA.

Conclusion
In addition to Tfh cell, the CXCR5−ICOShiPD-1hi CD4 T cell that produces IL-21 was increased in line with induction of SLE. This novel CXCR5−ICOShiPD-1hi CD4 T cell population appears to be responsible for helping B cell to induce varieties of autoantibodies and driving CD8 T cell to cause lupus tissue injury.
Proteomic Analysis Reveals Distinctive Protein Profiles Involved in CD8 T Cell-Mediated Murine Autoimmune Cholangitis
Authors: Zhang Weici , Zhang Ren , Zhang Jun , Sun Yin , Leung Patrick , Yang Guo-Xiang , Shuai Zongwen , Ridgway William , Gershwin M Eric 
Published: January 2023

Abstract
Autoimmune cholangitis arises from abnormal innate and adaptive immune responses in liver and T cells are critical drivers in this process, but little is known about the regulation of their functional behavior during disease development. We previously reported that mice with a T cell-restricted expression of a dominant negative form of transforming growth factor beta receptor type II (dnTGFbRII) spontaneously develop autoimmune cholangitis that resembles human primary biliary cholangitis (PBC). Adoptive transfer of CD8+ but not CD4+ T cells into Rag1−/− mice reproduced the disease, demonstrating a critical role for CD8 T cells in pathogenesis. Herein, we took advantage of SOMA scan technology to perform proteomic analysis of serum samples from dnTGFbRII and B6 control mice at different ages. In addition, we analyzed CD8 protein profiles after adoptive transfer of splenic CD8+ cells into Rag1−/− recipients. The use of the unique aptamer technology of SOMA scans reveals critical distinct profiles of CD8 cells, which are key to biliary mediation. 254 proteins were significantly increased, while 216 proteins significantly decreased in recipient hepatic CD8+ cells compared to donor splenic CD8+ cells. In contrast to donor splenic CD8+ cells, recipient hepatic CD8+ cells express distinct protein profiles involved in chemokine signaling, focal adhesion, T cell receptor and natural killer cell mediated cytotoxicity pathways.
Transcriptomics analysis reveals new insights into the roles of Notch1 signaling on macrophage polarization
Authors: Hans Chetan Parkash , Sen Sidharth , Zeng Shuai , Sharma Neekun , Dev Rishabh , Hans Ranjana , Joshi Trupti 
Published: January 2023

Abstract
Naïve macrophages (Mϕ) polarize in response to various environmental cues to a spectrum of M1 or M2 macrophages with distinct biological functions. Our earlier in vivo studies demonstrated that Notch1 deficiency attenuates the development of abdominal aortic aneurysm by promoting TGF-β2 dependent M2-polarization, however, the underlying mechanism is not completely understood. The present studies aimed to determine the specific roles for Notch1 signaling in macrophage polarization. Bone marrow derived macrophages isolated from WT or Notch1+/− mice (n=12) were differentiated to Mϕ, M1 or M2-phenotypes by 24h exposure to vehicle, LPS/IFN-γ or IL4/IL13, respectively. Total RNA isolated from these cells was subjected to RNA-Sequencing in triplicate. All samples had a minimum of 51.3 million 50 nucleotide paired-end reads and aligned at 87.5% ± 0.5% to the reference genome. EasyRNAseq and WebGestalt analysis with log2 fold change demonstrated that Notch1 haploinsufficiency downregulated the expression of 262 genes at basal level, 307 genes with LPS/IFN-γ and 254 genes with IL4/IL13 treatment. Among these, 50 unique genes including fibromodulin (Fmod), caspase-4, Has1, Col1a1, Alpl and Igf were downregulated by Notch1 haploinsufficiency under all the conditions. Pathway analysis demonstrated that extracellular matrix, macrophage polarization and osteogenesis to be the major pathways affected by Notch1 haploinsufficiency. Gain and loss-of-function and mechanistic studies established a strong correlation between Notch1 haploinsufficiency and Fmod in regulating TGF-β signaling. Overall, our studies provide strong evidence that Notch1 haploinsufficiency increases M2 polarization through these newly identified genes.