bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunoprotective γδ T cell subsets in the cornea during early HSV-1 infection.
Authors: Fitzpatrick Steffani E. , Lausch Robert , Barrington Robert A. 
Published: January 2023

Abstract
Herpes Simplex Virus-1 (HSV-1) is the leading cause of infectious blindness in the U.S.A., with an annual incidence of new ocular infections of 11.8 per 100,000. Both mice and humans develop similar pathology after intracorneal (i.c) HSV-1 infection, making mice a useful model to understand underlying mechanisms of disease. Mice lacking γδ T cells exhibit reduced Interleukin 17A (IL-17A), increased virus titers and develop lethal encephalitis after i.c. infection. Therefore γδ T cells are required for immunoprotection against i.c. HSV-1. To understand the diversity of the γδ T cell response to corneal infection, we examined corneal γδ T cell subpopulations based on C-C chemokine receptor 6 (CCR6) levels, IL-17A production, and the presence of activation markers, CD44 and CD62L. Both CCR6+ and CCR6neg γδ T cells were observed in the cornea. The number of CCR6+ γδ T cells increased within the first 48 hours post infection (p.i.). In addition, the CCR6+ γδ T cell subset were the predominant producers of IL-17A and possessed an activated cell phenotype (CD44hiCD62Lhi), consistent with central memory T cells.
To test whether the CCR6+ γδ T cell population is responsible for immunoprotection in the cornea, we measured corneal pathogenesis in HSV-1 infected CCR6 deficient (CCR6−/−) mice. CCR6−/− mice had significantly higher corneal pathology scores at days 3 and 7 p.i. compared to wild-type mice after HSV-1 i.c. These data suggest, that CCR6+ γδ T cells produce IL-17A and limit early corneal pathology.
Structural and Antigenic Features of Powassan Virus Envelope Protein
Authors: Errico John M. , VanBlargan Laura A. , Nelson Christopher A. , Diamond Michael S. , Fremont Daved H. 
Published: January 2023

Abstract
Flaviviruses, such as West-Nile virus and Dengue virus, are insect-transmitted positive-sense RNA viruses that cause substantial morbidity and mortality worldwide. Powassan virus is an emerging encephalitic tick-borne flavivirus endemic to the northern United States, and is currently the only tick-transmitted flavivirus known to infect humans in North America. In cases of severe neurological disease, up to 10% of patients die of Powassan encephalitis, while many survivors are left with long-term neurological sequelae. Despite this, no vaccines or therapeutics are currently available to treat Powassan virus infection. Flaviviral vaccination efforts are commonly foiled by an inability to generate potent neutralizing responses, and by genesis of cross-reactive antibodies that paradoxically enhance infection by heterologous flaviviruses. These neutralizing antibodies, and their cross-reactive counterparts, commonly target structurally homologous epitopes present on many flaviviruses. Thus, understanding these epitopes has important implications for vaccine design. Little is known about the structure of Powassan virus envelope protein or host antibodies that target it. We have recombinantly expressed Powassan virus envelope protein and are currently working to determine its structure. As well, a panel of Powassan-virus specific monoclonal antibodies has been developed. Utilizing our recombinant protein, we are characterizing the epitopes these antibodies utilize as well as their biochemical and functional properties. These findings will aid vaccine and therapeutic design for Powassan virus and expand our understanding of the quintessential antigenic and structural features of flaviviral envelope proteins.
An in vitro/ex vivo human lung infection model for studying the macrophage innate immune response to Ebola virus Kikwit
Authors: Josleyn Nicole M. , Kuehne Ana , O’Brien Cecilia , Brannan Jennifer , Edwards Darin , Parkhill Robert , Dye John M. 
Published: January 2023

Abstract
Ebola virus causes a severe and often fatal hemorrhagic fever in people, for which no approved vaccines or therapeutics are available. Disease pathogenesis and countermeasure studies are typically performed using animal models; however, ex vivo models recapitulating human immune responses may provide valuable information. We optimized engineered lung tissue constructs comprised of human lung epithelial cells and human endothelial cells that mimic the vascularized lower airway alveolar tissue micro-environment to study the human innate immune response to Ebola virus Kikwit (EBOV/Kik). Human lung tissue macrophages are difficult to obtain due to practical and ethical considerations, so macrophages were chemically derived from human monocytes. We inoculated the constructs, in the presence and absence of monocyte derived macrophages (MDM), and quantified cytokines/chemokines secreted on days 0, 3, 6, 9, and 15 post-infection using 29-plex and 1-plex human cytokine/chemokine kits. Prior to infection, MDM were secreting IL-13, MCP-1, MIP-1a, MIP-1b; however, these levels rapidly declined by day 3 post-infection (pi). The MDM secreted peak levels of IL-4, IFN-a2, Eotaxin, IL-12P40, and IFNg at day 6pi, and IL-1ra peaked at day 9 pi. In response to EBOV/Kik, the human lung epithelial cells in the engineered lung tissue construct secreted G-CSF, IL-1ra, IL-8, IP-10, and PDGF-AA/BB. G-CSF and IL-8, cytokines known to mobilize hematopoietic progenitors from bone marrow to blood and recruit neutrophils to sites of infection, were abundantly secreted by both endothelial cells and macrophages in response to infection. This model system may be a valuable resource to the field to study human immune responses during acute infections.
Antibodies that bind to an EBV derived peptide mimic of dsDNA, have pathogenic potential
Authors: Spatz Linda A. , Sarbaugh Dylan , Singh Divya , Kryvda Olga , Oudit Omar 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of antibodies to double stranded DNA (dsDNA) that can deposit in the kidney leading to nephritis. Our laboratory has previously been studying the association of SLE with the Epstein Barr Virus (EBV). We generated two monoclonal antibodies designated 3D4 and 16D2, that bind to a peptide (PFM-15) in the carboxyl region of a major nuclear protein of EBV (EBNA-1) and cross-react with dsDNA. In the present study, we demonstrate that both of these monoclonal antibodies can bind to fixed preparations of rat glomeruli suggesting pathogenic potential. Immunostaining is dramatically reduced when glomeruli are pre-treated with DNase indicating that these antibodies recognize chromatin associated structures. In addition, the PFM-15 peptide inhibits 3D4 and 16D2 from binding to glomeruli and inhibits both antibodies from binding to dsDNA coated plates by ELISA. This suggests that PFM-15 can act as a molecular mimic of dsDNA. Our results demonstrate that antibodies that recognize a viral peptide and cross-react with dsDNA can bind to glomeruli and therefore have pathogenic potential. This study further supports an association between EBV and SLE and supports the role of molecular mimicry in autoimmune disease.
Exposure to the Epstein-Barr viral antigen induces affinity-maturated anti-myelin antibodies.
Authors: Lomakin Yakov Anatol’evich , Belogurov Alexey 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is an autoimmune chronic inflammatory disease of central nervous system (CNS). Cross-reactivity of neuronal proteins with exogenous antigens is considered as one of the possible mechanisms of MS triggering. Previously we showed that monoclonal myelin basic protein (MBP)-specific IgGs are cross-reactive with Epstein-Barr virus (EBV) protein LMP-1. Here we report evidence that exposure of mice to LMP1 may result in induction of myelin-reactive autoantibodies in vivo. Only part of such anti-MBP antibodies are cross-reactive towards LMP1 but majority is occurred as a result of epitope spreading. We suggest that chronic contact with viral antigen rather than multiple rapid exposures to it is more sufficient in inducing switch of B cells from viral to myelin antigen. Moreover, even in inbred animals being almost identical in terms of genome, such switch was observed only in 20% of animals, indicating that this evidence is occurred by a chance rather than systematically. Our findings provide novel insights into still enigmatic link between EBV infection and MS development, determining several criteria that are beneficial for induction of self-reacting antibodies on the background of viral infection.
Current study was supported by Russian Science Foundation grant #17-74-30019.
Plasticity and functionality of Th17 subsets in EAE
Authors: Butcher Matthew J. , Zhu Jinfang (Jeff) 
Published: January 2023

Abstract
Several studies have suggested that IFNγ+IL-17A+ (or Tbet+RORγt+) Th17-like cells may be the key players in Experimental Autoimmune Encephalomyelitis (EAE). However, direct evidence demonstrating the importance and functionality of Tbet+RORγt+cells is lacking. Therefore, we examined the generation and stability of Th17 subsets during EAE, and assessed the contributions of RORγt and T-bet to the functionality of these cells. Using Tbx21ZsGreenRorcE2CrimsonFoxp3Rfp/y reporter mice, we were able to isolate T-bet and RORγt single and dual expressors after EAE induction. In a passive EAE model, we observed that donor RORγt-expressing Th17 cells elicited EAE passively and either maintained their phenotype, expressed T-bet, or became non-canonical Th1 cells (expressing T-bet only) in the recipients’ CNS. By contrast, T-bet and RORγt dual expressors largely assumed a non-cannonical Th1 cell fate; while donor Tbet+RORγt− cells maintained their phenotype and failed to elicit EAE. To assess if sustained RORγt expression in Tbet+ Th17 cells is required for the pathogenesis of EAE, we immunized Tbx21CreRorcFl/Fl and RorcFl/Fl mice. Interestingly, Tbet-dependent Rorc knockout mice did not develop EAE symptoms, and Tbet+ Th17 cells were absent in the CNS and dLNs. To examine the importance of RORgt downregulation by T-bet, we generated T-bet and Runx motif mutations within the Rorc promoter via Crispr/Cas9. RorcΔTbet/Runx mice similarly failed to develop EAE symptoms. Together these data suggest that Th17 cell subsets are functionally plastic and that the balance between T-bet and RORγt is required for the functionality of Th17/Th1 cells in EAE.
Interleukin 27 signaling in T cells is essential for the development of type 1 diabetes
Authors: Ciecko Ashley E. , Foda Bardees M. , Mueller Kevin W. , Geurts Aron M. , Chen Yi-Guang 
Published: January 2023

Abstract
Human genome wide association studies have identified a 0.73 Mb region on chromosome 16 significantly linked to the development of type 1 diabetes (T1D) and IL27 is proposed to be the causal gene. IL27 encodes the p28 subunit of the heterodimeric cytokine interleukin 27 (IL-27). Human eQTL studies suggest a disease promoting role for IL-27 in the pathogenesis of T1D. However, the function of IL-27 in T1D has not been completely defined. The focus of this study was to test the role of IL-27 in the progression of T1D using the NOD mouse model. We generated NOD mice deficient in IL-27 or IL-27 receptor (IL-27Rα). In sharp contrast to wildtype NOD mice, both NOD.Il27−/− and NOD.Il27ra−/− strains were completely protected from T1D. Additionally, histological examination of pancreatic sections revealed that both NOD.Il27−/− and NOD.Il27ra−/− mice had significantly less insulitis compared to the wildtype NOD control. Furthermore, IL-27 production by myeloid antigen presenting cells was sufficient to drive T1D progression. Adoptive transfer of IL-27 deficient T cells into NOD.Rag1−/− recipients resulted in diabetes development indicating that IL-27 deficient mice still harbor diabetogenic T cells. However, NOD.Rag1−/− recipients of IL-27Rα deficient T cells did not develop diabetes demonstrating that direct IL-27 signaling is essential for their diabetogenic activity. We further demonstrated that direct IL-27 signaling promoted the accumulation of CD8 T cells and altered CD4 T cell subsets (reducing Foxp3+ but increasing T-BET+ frequency) in pancreatic islets. Our observations reveal that IL-27 signaling in T cells is required for diabetes development in the NOD mouse and provide additional evidence to support its potential role in human T1D.
Understanding the role of STAT4 in experimental autoimmune encephalomyelitis
Authors: Anderson Ashlyn E. , Shin Boyoung , McWilliams Ian , Harrington Laurie E. 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is an autoimmune disease that affects over two million people worldwide. MS is characterized by the demyelination of axons in the central nervous system (CNS), leading to vision problems, muscle weakness and poor coordination. Among the various immune cells that contribute to the disease, a subset of CD4 T cells, Th17 cells has been associated with MS pathogenesis. Importantly, IL-23 signaling in Th17 cells is essential for the pathogenicity of these cells. A transcription factor downstream of IL-23 signaling, STAT4, is dispensable for in vivo and in vitro Th17 development, however STAT4 is crucial for the development of CNS inflammation and polymorphisms in STAT4 are linked to MS susceptibility. As STAT4 deficiency does not prevent Th17 differentiation, but still limits inflammation, we hypothesize that STAT4 is necessary for the pathogenic properties of Th17 cells in MS. Using the adoptive transfer model of experimental autoimmune encephalomyelitis (EAE), we find that STAT4 expression by Th17 cells is necessary to induce CNS inflammation. Furthermore, global gene expression analysis indicates that STAT4 regulates the recently described pathogenic and nonpathogenic Th17 gene signatures. In the absence of STAT4, the levels of pathogenic Th17 genes including Tbx21, Il22 and Cxcl3 are significantly reduced, while the expression of nonpathogenic genes including Il10 and Ahr is increased. Interestingly, we find that STAT4 influences Th17 gene expression regardless of the differentiation conditions. Together, these data reveal a novel role of STAT4 in controlling Th17 pathogenicity, which may provide a promising therapeutic target for MS patients.
Non-coding RNA regulation of immunity
Authors: Schaefer Jeremy , Schnupp Trevor 
Published: January 2023

Abstract
Non-coding RNAs (ncRNAs) comprise a diverse range of biologically functional RNA species that include long ncRNAs (lncRNA) and short ncRNAs such as piwi-interacting RNAs (piRNA) and microRNAs (miRNA). These ncRNAs are capable of regulating gene expression via distinct mechanisms depending on their structure---miRNAs bind specific messenger RNAs (mRNA) to block translation while lncRNAs tend to bind chromatin to modify chromatin states to interfere with gene expression. NcRNAs have been identified in numerous diseases as having altered expression. To further elucidate the role of ncRNAs in immunity, we conducted a screen of lncRNA expression in the interleukin-10 knockout mouse (IL-10−/−) model of intestinal inflammation. This screen identified several candidate lncRNAs with dysregulated expression. Importantly, our data demonstrate that Malat1 and Hoxa11as expression was significantly decreased in mice with severe intestinal pathology. These results suggest that an association exists between these lncRNAs and the progression of disease in the IL-10−/− mouse model of colitis.
IL-1α facilitates the entry of immune cells into the brain during chronic Toxoplasma gondii infection
Authors: Batista Samantha , O’Brien Carleigh , Harris Tajie H. 
Published: January 2023

Abstract
Toxoplasma gondii is a parasite that causes chronic infections in the brain. Infection is asymptomatic in healthy hosts, but immunocompromised hosts experience severe disease in which fast-replicating parasites destroy brain tissue. Constant immune pressure is required to maintain control of the parasite, leading to the localized recruitment of immune cells. During chronic T. gondii infection, cell death occurs in focal areas of inflammation in the brain, suggesting the possibility of inflammatory molecule release. The alarmin IL-1α is expressed in the brain, and is upregulated by T. gondii infection. IL-1α is expressed by brain-resident microglia and infiltrating macrophages, implicating these cell types as sources of this cytokine. Mice lacking IL-1α or its receptor (IL-1R1), but not IL-1β, have defects in macrophage infiltration into the brain during chronic infection, as well as increased parasite burden in the brain. IL-1α has been the subject of less investigation than IL-1β, but may still rely on inflammatory cell death for its release. Indeed, this IL-1α-dependent immune response appears to be dependent on pyroptosis, as caspase-1/11−/− mice phenocopy IL-1R1−/− mice displaying decreased recruitment of myeloid cells into the brain and increased parasite burden. IL-1R1 is expressed on endothelial and other stromal cells in the brain vasculature, and IL-1R1−/− mice display decreased expression of the adhesion molecules ICAM-1 and VCAM-1 in the brain. These data suggest that during chronic T. gondii infection, IL-1α released from a macrophage population in the brain through pyroptosis acts on brain vasculature to modulate expression of adhesion molecules, facilitating the infiltration of monocytes to sites of infection.
Cox10-deficient NK cells have increased apoptosis during MCMV infection, leading to susceptibility
Authors: Mah Annelise Y. , Keppel Molly P. , Saucier Nermina , Sexl Veronika , French Anthony R. , Cooper Megan A. 
Published: January 2023

Abstract
Studies in diverse cell types have rediscovered the importance of basic cellular metabolism in regulating immune responses. For example, glycolytic and oxidative metabolism are counter-regulated in T cells to develop inflammatory effectors vs. memory and regulatory cells, respectively. NK cells, which are first responders to viral infection and malignant transformation, may be similarly regulated. We recently showed that NK cell cytotoxic activity during infection with murine cytomegalovirus (MCMV) required glucose metabolism, whereas other NK cell responses such as cytokine production did not. We hypothesized that oxidative metabolism would be required for some of these glycolysis-independent responses. To test this, we examined responses to MCMV in mice with an NK-specific deletion of Cox10, which is required for assembly of electron transport chain complex IV. These “Ncr1-Cox10” mice had normal NK cell development and numbers, and stimulated Cox10-deficient NK cells survived and proliferated normally in vitro. However, Ncr1-Cox10 mice were more susceptible to MCMV infection, with impaired expansion of MCMV-specific Ly49H+ NK cells and increased apoptosis of splenic NK cells. Thus, we find that Cox10 is required by NK cells to expand and survive in response to MCMV infection. These deficiencies were not found in vitro, emphasizing the importance of using NK-specific genetic models to investigate metabolism in physiological contexts.
γδ T cells orchestrate the induction of protective Type 2 immunity in neonatal influenza infection
Authors: Guo Xizhi , Thomas Paul G. 
Published: January 2023

Abstract
Influenza is a cause of hospitalization and mortality in infants. γδ T cells are the first T cells to develop during embryogenesis and are a significant component of the neonatal immune system. In the neonatal period, conventional αβ T cell responses are functionally less prominent. γδ T cells play critical regulatory and effector roles, but their relevance in neonatal influenza remains to be investigated. In our neonatal infection model, we found γδ T cells were protective against mortality associated with neonatal influenza infection. Infection induced the accumulation of γδ T cells, which transiently expressed IL-17a, rather than IFN-γ. Subsequently, this led to stronger Type 2 immune responses than the TCRδ-deficient animals, particularly in IL-33 production and amphiregulin-mediated tissue repair. Loss of γδ T cells did not alter viral clearance or conventional anti-viral responses. We also observed an association between IL-17a levels and clinical outcomes. Thus, our results identify a specific requirement for γδ T cells in influenza-infected neonates for initiating Type 2 immune responses, mediating tissue homeostasis, and promoting lung integrity, which may provide a new therapeutic target for pediatric influenza treatment.
NOD2-mediated downregulation of the TLR pathways contributes to protection against colitis and colorectal tumorigenesis
Authors: Nashir Udden S. M. , Peng Lan , Zaki Hasan 
Published: January 2023

Abstract
NOD-like receptor family member NOD2, which activates the NF-kB and MAPK signaling pathways in response to the bacterial cell wall component MDP, is the first identified inflammatory bowel disease (IBD) susceptibility gene. IBD is a risk factor for colorectal tumorigenesis. However, the role of NOD2 in colorectal tumorigenesis has not been studied adequately and the precise mechanism of NOD2-mediated protection against IBD is elusive. We studied the role of NOD2 in colorectal tumorigenesis using azoxymethane plus dextran sulfate sodium-induced colorectal tumorigenesis in mice. We found that Nod2-deficient mice are highly susceptible to colorectal tumorigenesis with increased tumor burden as compared to wild-type mice. Interestingly, increased tumorigenesis in Nod2−/− mice was independent of gut microbial composition. Increased tumorigenesis in Nod2-deficient mice was associated with higher inflammatory responses and activation of the inflammatory signaling pathways, such as NF-kB, ERK, and STAT3. Consistently, in vitro studies demonstrate that while NOD2 activates NF-kB and ERK in response to MDP, it inhibits TLR-mediated activation of NF-kB and ERK, and production of inflammatory mediators. Notably, NOD2-mediated downregulation of TLR signaling pathways is independent of MDP-mediated activation of NOD2. We further explored the underlying mechanism of NOD2-mediated downregulation of TLR signaling and revealed that NOD2 induces IRF4 which negatively regulates TLR-NF-kB signaling axis. Taken together, this study therefore shed light on a novel function of NOD2 which may be implicated in genetic association of NOD2 mutations with IBD and colorectal cancer.
Nucleic acid sensing in the cytoplasm of C2C12 cells directly after plasmid DNA electroporation
Authors: Semenova Nina P. , Znidar Katarina , Bosnjak Masa , Cemazar Maja , Heller Loree C. 
Published: January 2023

Abstract
Mounting evidence stresses the importance of nucleic acid sensing in the cytoplasm of living cells in a variety of physiological and pathological processes, including anti-pathogen defense, autoimmunity and development of malignant tumors. A subset of pattern-recognition receptors (PRRs) are nucleic acid sensors widely expressed in a variety of cells, but the detailed mechanisms of nucleic acid sensing have not been clarified. Our previous studies confirmed the cell type-specific upregulation of PRRs on both the mRNA and protein levels after electroporation of plasmid DNA (pDNA) into the cells. Type I interferon was found to be upregulated as well, which implicates innate immunity pathway activation in response to pDNA entry into the cytoplasm. Still, the sequence of events after pDNA electroporation remains unclear. Using molecular and proteomic techniques, we identified DAI/ZBP1 protein as the initial responding PRR to pDNA entering the cytoplasm after electroporation and estimated the participation of other PRRs in this process in mouse myoblast C2C12 cells. Better understanding of molecular determinants of nucleic acid sensing by PRRs may allow us to manipulate this process, developing new and improving existing therapies for immunity- and tumor-related diseases.