bluuebunny/crossref_metadata_2025_split huggingface dataset:
Maturation of dendritic cells derived from type 1 diabetes-susceptible NOD mice is differentially regulated by two strains of generally regarded as safe (GRAS) Lactobacillus
Authors: Larkin Joseph , Mohamed Zaynab Sidi , Ardissone Alexandria , Bedoya Simone , Collins Erin , Wasserfall Clive , Lorca Graciela , Triplett Eric W. , Atkinson Mark 
Published: January 2023

Abstract
Although commensal bacteria have been implicated in type 1 diabetes (T1D), specific mechanistic insight is limited. We hypothesized that specific strains of commensal bacteria can modulate the diabetogenic peripheral T cell effector repertoire (PTER) through differential dendritic cell (DC) maturation. To test this, we assessed the presence of an IL17+ PTER and compared the microbial profiles present within female non-obese diabetic (NOD) mice, which spontaneously develop TID, and non-obese resistant (NOR) mice, which are resistant to T1D. We also assessed changes in DC maturation mediated by generally regarded as safe (GRAS) Lactobacillus johnsonii N6.2 (LjN6.2) versus Lactobacillus reuteri TD1 (LrTD1), which were previously shown to inhibit and accelerate T1D, respectively, in bio-breeding diabetes-prone (BBDP) rats. We found that NOR mice possessed a distinct Th17 bias and higher CD11c+MHCII+ DCs frequencies within lymph nodes and pancreas when compared to NOD mice. In vitro, LjN6.2 induced production of IL-6, IL-10, and TGF-b, previously shown to prime protective IL17+ T lymphocytes. Additionally, LjN6.2 promoted dendritic cell expression of the tolerogenic signaling molecules SOCS1, B7-H4, and PD-L1. In contrast, LrTD1 promoted production of proinflammatory cytokines, IL-12 and IL-23, which share a common p40 subunit. Together these data suggest that certain bacterial strains may enhance tolerance mechanisms and, thereby, contribute toward an environment that might protect against T1D, whereas bacterial strains such as LrTD1 may act as an “inducing” environmental factor by enhancing IL-12 production. Hence, these results emphasize a need to elucidate immunological mechanisms mediated by GRAS bacteria.
TLR9 stimulation by the gut microbiota promotes tissue-specific immunosuppression in autoimmune-prone MRL/lpr mice
Authors: Edwards Michael Richard , Mu Qinghui , Swartwout Brianna , Cabana-Puig Xavier , Mao Jiangdi , Reilly Christopher M. , Luo Xin M. 
Published: January 2023

Abstract
To investigate the ability of gut microbiota to modify immune regulation in autoimmune disease, we treated lupus-prone mice with oral vancomycin prior to disease development, and subsequently administered bacterial DNA isolated from commensal bacteria. Vancomycin given prior to disease initiation promotes exacerbation of autoimmune disease, likely due to a decrease in the frequency of B regulatory (Breg) cells, whereas vancomycin given after autoimmune disease onset has suppressive effects on disease phenotype in MRL/lpr mice. Bacterial DNA administration during the pre-disease stage promoted Breg cell production and suppressed autoimmune disease parameters in MRL/lpr mice. Further, we showed that activation of toll-like receptor 9 (TLR9) on B cells was involved in the suppressive effects of bacterial DNA seen in the MRL/lpr mouse model. We found that during the pre-disease period, joint treatment with vancomycin and oral administration of a TLR9 specific agonist, ODN 1826, promoted Breg cells in the lamina propria, increased total CD8+ T cells and effector memory CD44 expressingCD8+ T cells in the spleen, and suppressed Th17 and IFNγ producing CD4+ T cells in the mesenteric lymph nodes. Oral ODN 1826 administration also increased the Treg/Th17 ratio in the mesenteric lymph nodes. The alterations in suppressive vs. proinflammatory cell phenotypes by oral exposure to ODN 1826 suggests that the suppressive effects of bacterial DNA from the gut microbiota are modulated through TLR9 activation in a tissue-specific manner.
Efficacy of cannabidiol treatment in experimental MS is due to immunosuppressive activity of myeloid cells in CNS downregulating recruitment of CD4+ T cells, proinflammatory chemokines and gasdermin D expression
Authors: Dopkins Nicholas , Wilson Kiesha , Miranda Kathryn , Nagarkatti Prakash S , Nagarkatti Mitzi 
Published: January 2023

Abstract
Cannabidiol (CBD) is a nonpsychoactive ingredient from Cannabis that has garnered attention in the medical community due to its anti-inflammatory properties and therapeutic potential. CBD has in particular become a popular alternative medicine among individuals with autoimmune disorders due to CBD lacking the side effects and costs associated with immunosuppressants. To better define how CBD inhibits inflammation, we studied the effects of orally administering CBD (20mg/kg) in experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis. Using single cell RNA sequencing on cells isolated from the central nervous system (CNS) of EAE mice, we demonstrated that CBD treatment inhibits neuroinflammation by regulating gene expression and infiltration of myeloid cells. Within the resident and infiltrating myeloid cells of the CNS, there was decreased expression of inflammatory cytokines and chemokines (CXCL9, CXCL10, IL-18) associated with neuroinflammation and CD4+ T cell recruitment, pyroptosis initiators (gasdermin D; Gsdmd), oxide synthesizers (Arg1) and antigen presentation mediators (CD40) in CBD-treatment group. Additionally, CBD yielded an increase in the polymorphonuclear myeloid derived suppressor cells in the CNS, signifying immunotolerance induction at the site of inflammation. In the VEH treated mice, we found CD4+ T cells that express the receptor for IL-18 (IL-18r1) that are absent in CBD treated mice, suggesting CBD affects T cell activity via inhibition of myeloid cell activity. This data shows that CBD treatment ameliorates EAE by inhibiting infiltration, pro-inflammatory cytokine production, antigen presentation, T cell recruitment and pyroptosis of myeloid cells in the CNS.
Exhausted CD8 T cells exhibit phenotypic and functional heterogeneity in type I diabetes (T1D)
Authors: Kus Anna , Wiedeman Alice , Wall Valerie , Muir Virginia , Speake Cate , Long S. Alice 
Published: January 2023

Abstract
CD8 T Cell exhaustion is associated with poor clinical outcome in cancer and chronic infection, but positive outcome in autoimmunity. Recently, phenotypic and functional heterogeneity within exhausted CD8 T cells suggests lineage subsets. We observed similar heterogeneity in the phenotype of exhausted cells in the context of T1D. Here we further explore the phenotypic and functional features of exhausted subsets, and ask about correlation with features of T1D. Using a CyTOF panel, we identified 12 major phenotypes (clusters) among total CD8 T cells in T1D subjects. Four of these clusters were characterized by features of exhaustion, namely PD-1, EOMES, KLRG-1, and TIGIT expression. Of these four, a CD127 high, but Granzyme B and TBET low cluster was enriched for islet-specific autoreactive CD8 T cells, and was associated with slower disease progression. As compared with other exhausted clusters, CD127-high exhausted cells proliferated more and produced increased IFNγ and TNFα upon in vitro anti-CD3 stimulation, measured by flow cytometry. This phenotype and functionality is consistent with a stem-like exhausted population. In comparison, the other three other clusters expressed higher TBET and Granzyme B but lower CD127, two of which were enriched for insulin-specific cells. This is consistent with a transitory exhausted phenotype driven by repeated antigen exposure as occurs with insulin treatment of T1D. Thus, varying stages of exhaustion are present in T1D and the stem-like exhausted phenotype may be beneficial in slowing disease progression in T1D. Understanding exhausted CD8 T cell heterogeneity and the role these subsets play in T1D will lead to improved therapies.
A novel self-regulatory mechanism of Th17 cells controls autoimmune uveitis through interleukin-24
Authors: Chong Wai Po , Mattapallil Mary , Raychaudhuri Kumarkrishna , Silver Phyllis B. , Jittayasothorn Yingyos , Chan Chi-Chao , Chen Jun , Horai Reiko , Caspi Rachel R 
Published: January 2023

Abstract
The Th17 response is critical in driving inflammation and is associated with many autoimmune diseases. However, clinical trials targeting the Th17 signature cytokine, IL-17A in some autoimmune diseases, including uveitis, have been disappointing. We investigated the role of IL-17A in determining the pathogenicity of uveitogenic Th17 cells. Unexpectedly, IL-17A deficiency in these cells did not reduce the severity of uveitis in recipient mice. We found that IL-17A deficient Th17 cells produced elevated amounts of other Th17-related cytokines, i.e. IL-17F, GM-CSF and IL-22. RNA-seq analysis and the follow up in-vitro experiments revealed that IL-17A exerts a negative feedback on Th17 cells by inducing them to produce IL-24, which in turn suppresses the production of Th17-related cytokines in an autocrine fashion. In vivo results showed that IL-24 treatment of recipients ameliorated Th17-induced adoptive EAU, and conversely, silencing IL-24 expression in retina-specific Th17 cells increased their pathogenicity, supporting the relevance of this pathway in vivo. Mechanistic studies confirmed that IL-17A activates the NFkB signalling pathway in Th17 cells, whereas site mutagenesis at the NFkB binding sites In IL-24 promoter abrogates the IL-17A-induced IL-24 expression. In vitro experiments implicated SOCS 1 and 3 in the downstream effects of IL-24. Importantly, we also observed a similar inhibitory IL-17A/IL-24 circuit in polarized human Th17 cells. We conclude that IL-17A exerts a negative feedback on Th17 cells by inducing autocrine IL-24, which limits their expression of other Th17-lineage cytokines and dampens their pathogenicity.
LAG3 limits the diabetogenicity of intra-islet CD8 T cells
Authors: Grebinoski Stephanie J , Zhang Qianxia , Cillo Anthony R , Szymczak-Workman Andrea , Workman Creg , Vignali Dario 
Published: January 2023

Abstract
Tight control of effector T cells is necessary to limit autoimmune diabetes (AD). One mechanism by which this is achieved is through inhibitory receptor (IR) expression. Lymphocyte Activating Gene 3 (LAG3) is one critical IR that limits disease progression and incidence of AD. Using a mouse model in which LAG3 deletion is restricted to CD8+ T cells, Lag3L/L-YFPE8iCRE-GFP.NOD, and wild type (WT) controls, E8iCRE-GFP.NOD, we find that CD8+ T cell-restricted deletion of LAG3 is sufficient to accelerate diabetes incidence and insulitis. Interestingly, we see Lag3L/L-YFPE8iCRE-GFP.NOD mice have an increased intra-islet CD8:Treg ratio, and increased tetramer (Nrpv7, IGRP mimotope) staining. These observations drive our hypothesis that LAG3 limits those autoreactive CD8+ T cell. 5′ single cell RNAseq in conjunction with TCRseq reveals that LAG3-deficient (Lag3−/−) intra-islet CD8+ T cells are transcriptionally unique compared to WT controls, exhibiting a more activated phenotype along with lower geneset enrichment for markers of exhaustion. This was validated by flow cytometry in which WT CD8+ T cells possess an expanded PD1Hi, Tox+ population, while Lag3−/− CD8+ T cells have an expanded PD1mid population, express more CD44, KLRG1, BrdU, Ki67, and less aCasp3. This indicates that WT CD8+ T cells are able to appropriately upregulate PD1 and Tox in response to chronic self-antigen stimulation, while Lag3−/− CD8+ T cells maintain a more activated population. Strikingly, LAG3 deletion on CD8+ T cells appears to be sufficient to overcome a normal “exhaustion-like” program in intra-islet CD8+ T cells, resulting in accelerated diabetes incidence. These data highlight an important role for LAG3 and CD8+ T cell exhaustion programs in AD.
Dissection of T-cell tolerance induction by dendritic cell surface glycoproteins
Authors: Dustin Michael , Saadawi Ahmed , Mayya Viveka , Cespedes Pablo , Valvo Salvatore , Chen Lina , Demetriou Philippos 
Published: January 2023

Abstract
Tolerogenic dendritic cells (DC) induce T-cell clonal deletion, anergy, generation and activation of regulatory T cells. In vitro production of TolDC has been accomplished, however the full molecular circuitry is not known. We aim to decipher the chemical signals and physical characteristics leading to T-cell tolerance in an artificial system. We have performed surface proteomic analysis of human peripheral blood cDC2 subset without and with in vitro stimulation with polyIC in vitro. The surface proteomics is based on biotinylation of sialic acids and the signal strength was 55-fold higher in the resting compared to the activated condition, presumably due to decreased sialation of glycoproteins in the activated cDC2. We found that label free quantification (LFQ), corrected for differences in sialation, agreed well with quantitative surface expression by flow cytometry. We combined the two datasets and ranked the top 120 glycoproteins with extracellular domains (ECD) &gt; 60 amino acids by LFQ with approximately one third invariant, one third up-regulated/induced and one one-third down-regulated/lost on activation. The 120 ECD were expressed in HEK293 cells with C-terminal 10His tags and purified by Ni2+ affinity chromatography. Supported lipids bilayers (SLB) representing the tolerogenic and activating DC are being designed with different combinations of the 120 ECDs. We will adjust sialation to match resting and activated states. The SLB compositions will be tested for immunological synapse formation (fluorescent self-pMHC/CD80, ICAM-1 and CD58 for the cSMAC, pSMAC and dSMAC) and tolerance/activation spectrum with identification of relevant molecular combinations by machine learning.
UVB light exposure promotes IgE autoantibody production and skin autoreactivity in mice that express a lupus-associated isoform of Foxp3
Authors: Domeier Phillip P , Chang Sarah E , Spath Sabine , Utz Paul J , Zhou Baohua , Ziegler Steven F 
Published: January 2023

Abstract
Foxp3 activity is required for the maintenance of self-tolerance, but humans encode an additional exon 2-deficient splice variant isoform of FOXP3 with no known function. To determine how exon 2 of FOXP3 contributes to Treg activity, we generated mice that express exon 2-deficient FoxP3 (called B6.FoxP3ΔExon2). B6.FoxP3ΔExon2 mice develop mild autoimmunity (splenomegaly, autoantibodies and kidney nephritis) with significantly elevated titers of skin-reactive IgE. We also observed skin-reactive IgE antibodies in SLE patients with active photosensitivity, so we challenged B6 and B6.FoxP3ΔExon2 mouse skin with ultraviolet B (UVB) light to determine if UVB damage can induce skin-reactive IgE production. When compared to B6 controls, B6.FoxP3ΔExon2 mice developed greater skin inflammation with increased infiltration of Tregs (CD4+Foxp3+), effector T cells (CD4+CD44+Foxp3−) and IgE+ plasma cells (B220lowCD138+Blimp1+) at the site of UVB damage. B6.FoxP3ΔExon2 mice also had higher numbers of IgE+ antibody-forming cells, IL4-producing T cells (CD4+CD44+Foxp3−) and IgE-producing B cells, but lower numbers of CTLA4+ Tregs in draining lymph nodes from UVB-treated skin. Finally, UVB-treated B6.FoxP3ΔExon2 mice also had worsened kidney disease, phenocopying multi-systemic flares that occur in SLE patients with photosensitivity. Collectively, we found that exon 2 of the Foxp3 gene is required to control UVB-induced cutaneous inflammation though the regulation of type 2 immune activation and skin-reactive IgE. Thus, our findings suggest that photosensitivity reactions in SLE patients may be mediated by altered Foxp3 activity and skin-reactive IgE that could be therapeutically targeted to control systemic autoimmune flares.
Characterization of Aire-expressing DCs using a novel Aire-reporter strain
Authors: Matsumoto Mitsuru , Matsumoto Minoru , Miyazawa Ryuichiro , Morimoto Junko , Tsuneyama Koichi , Nishijima Hitoshi 
Published: January 2023

Abstract
Aire is predominantly expressed by medullary thymic epithelial cells (mTECs), and it is essential for maintaining self-tolerance by controlling the expression of tissue-restricted self-antigens (TRAs). Besides mTECs, extra-thymic Aire-expressing cells (eTACs) have been reported. However, the exact nature of eTACs has been difficult to study because of the small numbers together with low expression levels of Aire within the cells. We have recently established a novel Aire reporter strain in which endogenous Aire was replaced by the human AIRE-GFP-Flag tag fusion gene (Aire/hAGF-knockin: J. Immunol. 2015). hAGF-reporter protein was efficiently produced and retained within the mTECs as authentic Aire nuclear dot protein. With this high-sensitivity and high-fidelity Aire-reporter strain, we found that DCs in the spleen and lymph nodes expressed Aire although their expression levels were rather low. Aire-expressing DCs showed high levels of MHC-II and CD80/CD86, and half of which expressed CD8. In the thymus, we found that there were two types of DCs expressing Aire: hAGF-low DCs and hAGF-high DCs. Interestingly, hAGF-low DCs had small hAGF dots within the nuclei, whereas hAGF-high DCs retained larger hAGF dots which were mainly present outside the nuclei, suggesting that hAGF-high DCs had acquired hAGF protein from mTECs. Acquisition of hAGF protein by hAGF-high DCs from mTECs was confirmed by the bone-marrow transfer experiment. Mechanisms underlying the transfer of nuclear protein (i.e., hAGF protein) from mTECs to thymic DCs, and immunological consequence of this event are now under investigation.
Pathogenic role of plasmacytoid dendritic cells in the development and onset of Sjögren’s syndrome-like exocrinopathy in non-obese diabetic mice
Authors: Zhou Jing , Zhang Xiaofeng , Yoo Bora , Yu Qing 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDCs), which are the most potent producers of type I IFNs, have been demonstrated to be critically involved in the induction of multiple autoimmune diseases. In Sjögren’s syndrome (SS), the presence of pDCs and activated type I IFN pathways has been shown in the salivary glands of patients, and the disease severity closely correlates with type I IFN activity. However, the particular role of pDCs in the pathogenesis of SS remains unknown. To investigate whether pDCs play a crucial role in the development and onset of SS, we administrated a pDC-depleting anti-BST2 antibody to female non-obese diabetic (NOD) mice, a spontaneous model of SS during the early stage of SS development, from 4 to 7 weeks of age. Mice were analyzed at 10 weeks of age around the clinical onset of the disease. Anti-BST2 treated mice had significantly higher salivary flow rate than IgG-treated control group, indicating the remission of SS-like xerostomia by pDC depletion. Meanwhile, anti-BST2 treatment inhibited leukocyte infiltration of submandibular glands (SMGs), as demonstrated by H&E staining. Consistent with these results, flow cytometric analysis provided evidence that administration of anti-BST2 not only lowered the frequency of total leukocytes, CD4 T cells, CD8 T cells, B cells and pDCs, but also reduced the proportion of CD44+CD4 T cells, CD44+CD8 T cells and CD86+B cells among the total SMG cells of NOD mice. These changes were accompanied by downregulation of CD40, CD40L, Lt-α, Lt-β, TNF-α, IFN-γ, CXCL9, CXCL11 and NOS2, as determined by RT2 Profiler PCR array, suggesting an attenuation of SMG inflammation. Taken together, pDC depletion at the early stage of disease development hinders the pathogenesis of SS-like exocrinopathy.
Helminth-induced modulation of neuroinflammation in a mouse model of multiple sclerosis
Authors: Fettig Naomi M , Horwitz Marc S , Osborne Lisa C 
Published: January 2023

Abstract
Environmental factors play an important role in multiple sclerosis (MS) disease progression, but the exact cause remains unknown. The hygiene hypothesis suggests that a lack of exposure to microbes may alter disease susceptibility. Helminths in particular may play a role in preventing autoimmune disease due to the type 2 immune response they typically induce. Trichinella spiralis (Ts) first forms a transient intestinal infection followed by chronic infection where it exerts type 2 and regulatory immune responses, respectively. To evaluate the role of helminth infection in development and progression of MS, C57BL/6J mice were first chronically infected with Ts, then immunized with MOG35–55 to induce experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. Ts-infected EAE mice have an altered disease course compared to uninfected controls, demonstrating delayed onset, reduced incidence, and symptom remission. Despite an overall decrease in lymphocyte recruitment to the central nervous system (CNS) in Ts/EAE mice, elevated numbers of infiltrating Th2 cells are maintained through the EAE disease course. In contrast, regulatory T cell expansion is dependent on the presence or absence of EAE symptoms. Histological analysis of cellular infiltration and demyelination in the spinal cord indicate limited demyelination with reduced infiltration into the parenchyma of Ts/EAE mice compared to EAE controls. Further immunophenotyping in the CNS to evaluate the contributions of infiltrating macrophages will be performed, as well as analyses of the cytokine milieu in the CNS. These results suggest that chronic Ts infection limits neuroinflammation, as shown by reduced clinical, immunological, and pathologic signs of EAE.
Foxp3+ regulatory T cells play an indispensable role during glucocorticoid-induced treatment of autoimmune inflammation
Authors: Kim Dongkyun , Min Booki 
Published: January 2023

Abstract
Glucocorticoid (GC) is a class of steroid hormones with profound immunosuppressive and anti-inflammatory properties that is widely used as the first-line treatment option in chronic inflammatory conditions such as autoimmunity. Despite the broad usage, its working mechanisms remain largely unclear. Herein, we report that Foxp3+ CD4+ regulatory T cells (Tregs) play an irreplaceable role during GC-mediated treatment of experimental autoimmune encephalomyelitis (EAE), a murine autoimmune inflammation model in the central nervous system that resembles multiple sclerosis in human. Dexamethasone (Dex) administered at the disease onset or ongoing disease was unable to suppress EAE without Tregs, while adoptive transfer of Tregs restored Dex effects. Glucocorticoid receptor (GR) expression in Tregs was essential for Dex effects, suggesting that GC directly acts on Tregs. From RNA sequencing analysis we found that Dex treatment induced miR-342 expression only in Tregs but not in effector T cells. We also found that miR-342 targets the Rictor, a subunit of the mTORC2 complex. Inhibiting Dex-induced miR-342 expression in Tregs not only restored Rictor expression but also interfered with Treg-mediated suppression of EAE even with Dex treatment. While Dex-stimulated Tregs displayed robust oxidative phosphorylation rather than glycolysis as the energy source, miR-342 inhibition in Tregs reversed metabolic program to robust glycolysis. Taken together, this is the first report demonstrating that the miR-342-Rictor axis may represent a novel mechanism by which GC controls inflammation by metabolic programming in Tregs.