bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mycobacterium avium subspecies paratuberculosis (MAP) drives an innate Th17-like T cell response and upregulation of inhibitory signals
Authors: DeKuiper Justin Lee , Coussens Paul M 
Published: January 2023

Abstract
Johne’s disease (JD) is a chronic inflammatory gastrointestinal disorder of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP). Our work suggested that MAP may drive a non-classical Th17-like response in PBMCs. Our objectives were to determine if Th17 cell activation by MAP required the presence of antigen presenting cells (APCs) and to determine how epithelial cells might influence Th17 activation. Monocytes were isolated by adhering and washing protocols and then cultured for 5 days to allow differentiation into macrophages. T cells and B cells were isolated using magnetic-activated cell sorting (MACS) by positive selection of CD3+ and sIgM+ cells, respectively. MACS cells were then cultured or cocultured with macrophages. MDBK cells were used as an epithelial cell model. qPCR was used to determine mRNA expression levels of MDBK, T cells alone, and T cells cultured with macrophages or B cells. All T cell cultures increased IL-17a and IL-23 mRNA expression with MAP-antigen stimulation when compared to untreated cells. APC containing cultures expressed significantly more IL-17a than T cells alone. IL-22 expression was only observed in T cell cultures stimulated with live MAP. T cell exhaustion and tolerance markers (PD-1, PD-L1, LAG-3) were increased in T cell only cultures when stimulated with MAP, but repressed in the presence of APCs. The presence of MDBK cells upregulated IL-23 following stimulation with MAP, suggesting that epithelial cells exposed to MAP may also help drive Th17 differentiation.
Sargassum horneri mitigates particulate matter induced inflammatory response in MLE-12 cells via MAPK pathways
Authors: Mihindukulasooriya Suyama Prasansali , Herath Kalahe Hewage Iresha Nadeeka Madushani , Kim Areum , Kim Hyo Jin , Ko Mi-Ok , Jeon You-Jin , Youngheun Jee 
Published: January 2023

Abstract
Rise of ambient particulate matter (PM) exhibits positive correlation with increased respiratory diseases since PM triggers an inflammatory response in lung. As approaches aiming at limiting PM exposure provide only partial solutions, we investigated a therapeutic approach to mitigate harmful effects of PM in this study. Specifically, we examined the anti-inflammatory potential of the 80% ethanol extract of Sargassum honeri (SHE) against PM exposed murine lung epithelial cells (MLE-12). PM exhibited a cytotoxic effect on MLE-12 from rather low concentrations (concentrations &gt; 15.6 μg/mL; IC50 44.1 μg/mL), and the cytotoxic concentration of PM utilized in this study was 125 μg/mL. But, SHE (0–125 μg/mL) was not cytotoxic on MLE-12 cells, and concentrations 62.5 and 125 μg/mL were tested in this study. SHE reversed the PM induced lactate dehydrogenase (LDH) release and cell proliferation loss of MLE-12 cells. SHE also decreased the PM induced secretion of pro-inflammatory cytokines (IL-1β, TNF-α), pro-inflammatory mediator COX-2, anti-inflammatory cytokines (IL-6, IL-8) and inflammatory chemokines (MCP-1, CCL5). Furthermore, SHE decreased PM induced phosphorylation of p38 mitogen-activated protein (MAP) kinase, extracellular signal-regulated kinase 1/2 (Erk1/2) and c-Jun NH (2)-terminal kinase (JNK). Taken together, we conclude that SHE’s anti-inflammatory effect was regulated probably via the inhibition of p38, ERK and JNK MAP kinase signaling pathway in MLE-12 cells. This research is a part of the project, “Development of functional food products with natural materials derived from marine resources”, funded by the Ministry of Oceans and Fisheries, Korea in 2017 (Project NO. 20172085).
CCR2 mediates transendothelial migration of human pathogenic Th17 cells
Authors: Parween Farhat , Kathuria Noshin , Zhang Hongwei , Farber Joshua M 
Published: January 2023

Abstract
Leukocyte extravasation is characterized by rolling and arrest on the endothelium followed by transendothelial migration (TEM). Leukocyte arrest and TEM typically require chemokines and their receptors. Previously, we showed that CD4+CCR5+CCR2+ T cells are highly differentiated, readily activated memory cells in human blood that exhibit diverse effector capabilities. We and others have also shown that CCR6 is the signature chemokine receptor for Th17 cells. Here we describe studies of the migratory capabilities and effector profiles of CD4+CCR6+CCR2+ as compared with CD4+CCR6+CCR2− and CD4+CCR6−CCR2− effector-memory T cells from human blood. Using flow chamber assays with TNFa-activated human umbilical vein endothelial cells, we found that among the CCR6+ cells, only the CCR2+ subset was able to cross the endothelial monolayer efficiently. We found that CCR6 and CCR5 were important for arrest whereas CCR2 was critical for subsequent TEM. Our RNA-Seq data and intracellular staining data of ex vivo activated T cells showed that the CCR6+CCR2+ cells were distinguished by their expression not only of the cytokines typical of Th17-cells, but also of cytokines, including IFNg and GM-CSF, and transcription factors characteristic of the pathogenic Th17 cells that mediate tissue injury in mouse models of autoimmune disease. Taken together, our studies suggest that chemokine receptors on memory Th cells play non-redundant roles in the egress from blood, that providing T cells with a combination of CCR6 CCR5 and CCR2 could enhance their trafficking into tissue, and that, on the other hand, blocking CCR2 alone could prevent the extravasation of pathogenic Th17 cells and potentially diminish tissue damage in Th17-cell driven disease.
Intrinsic Alteration of Differentiation and Glycolysis in T Lymphocytes Immediately After TCR Activation Through PSGL-1 Signaling
Authors: Hope Jennifer L , Otero Dennis , de Jong Petrus , Ma Jiadai , Henriquez Monique , Powis Garth , Bradley Linda 
Published: January 2023

Abstract
P-selectin glycoprotein ligand-1 (PSGL-1) is an adhesion molecule expressed on the surface of naïve, effector, and memory CD4+ and CD8+ T cells. We identified PSGL-1 to be an immune checkpoint inhibitor as antibody-mediated ligation of PSGL-1 promotes T cell exhaustion and deletion of PSGL-1 prevents chronic viral infection and inhibits tumor. To investigate the role of PSGL-1 signaling in the development of T cell responses, we assessed the differentiation state, expansion capacity, and glycolytic profile of PSGL-1-deficient T cells. PSGL-1+/− OT-II CD4+ T cells demonstrated increased skewing towards IL-17A+ T cells compared to wild-type OT-II CD4+ T cells under TH17 conditions, and reduced IFNg+ cells under both TH0 and TH1 conditions. In a B16-OVA tumor model, we observed increased IL-13+ CD4+ T cells among the intratumoral T cell compartment in PSGL-1−/− mice. In vitro activation with a sub-optimal dose of αCD3 of PSGL-1−/− OT-II CD4+ T cells demonstrated increased expansion and expression of CD25 compared to wild-type OT-II CD4+ T cells. Further, adoptively transferred in vitro-activated PSGL-1−/− CD4+ T cells demonstrated greater expansion in vivo upon adoptive transfer into RAG−/− host mice. Using the Seahorse glycolysis stress test, we identified that both CD4+ and CD8+ PSGL-1−/− T cells demonstrate increased glycolysis after 72 hours of in vitro activation compared to wild-type T cells. Further, in situ activation of PSGL-1−/− CD8+ T cells demonstrates that at both sub-optimal and optimal levels of αCD3 stimulation, PSGL-1−/− CD8+T cells have increased glycolysis and increased glycolytic capacity. Taken together, these data show that PSGL-1 signaling has an intrinsic and immediate role in the development of T cell responses.
IL7R regulates fetal tissue resident macrophage development
Authors: Leung Gabriel , McCann Taylor , Valencia Clint H. , Worthington Atesh , Forsberg Camilla , Beaudin Anna E 
Published: January 2023

Abstract
Tissue-resident macrophages (TRMs) play critical roles in tissue homeostasis and disease. Many populations of TRMs derive from fetal progenitors and independently self-maintain across the lifespan through in situ proliferation. Here, we have identified the interleukin-7 receptor (IL7R) as a novel regulator of TRM development. Using an IL7R-Cre lineage tracing model, we observed that adult TRMs in the brain, epidermis, liver, and lung were highly labeled by IL7R-cre, in the absence of IL7Ra mRNA or protein expression. To gain insight into developmental expression of IL7Ra, we profiled surface expression, mRNA expression, and IL7R-cre driven labeling across fetal development. Erythromyeloid progenitors, putative TRM precursors, were barely labeled by IL7Ra-cre, and IL7R deletion did not affect YS hematopoiesis. In contrast, we observed IL7Ra mRNA expression in fetal monocytes, and robust IL7Ra surface expression on developing TRMs during late gestation. Sorted Ly6chi fetal liver monocytes cultured ex vivo with M-CSF differentiated into macrophages expressing IL7R, suggesting a precursor-product relationship. Blockade of the IL7R with a monoclonal antibody during gestation impaired liver, lung, and epidermal TRM cellularity at birth, with a concomitant increase in cellularity of liver monocytes, suggesting that IL7Ra regulates TRM differentiation from fetal monocytes during fetal development. These data reveal dynamic regulation of IL7Ra expression in TRMs and TRM precursors during late gestation, and provide evidence that IL7R signaling regulates fetal TRM development. Ongoing work addresses downstream signaling and the specific developmental processes regulated by IL7R signaling during fetal TRM development.
Bone marrow resident Tregs maintain stromal cell function via IL-10
Authors: Camacho Virginia , Patel Sweta , Matkins Victoria , Turnquist Heth R , Welner Robert S 
Published: January 2023

Abstract
Regulatory T cells within the bone marrow (BM Tregs) represent a specialized population whose functions are incompletely understood. In this study, we provide a novel mechanism by which BM Tregs support the maintenance of stromal cells to direct hematopoiesis. When compared to Tregs found in spleen and lymph node, they express genes associated with activation as well as chemokine receptors. As another distinguishing characteristic, BM Tregs highly express both CD49b, a marker of circulating Tregs, and S1PR1, a receptor involved in trafficking. This was corroborated by parabiosis and transplant experiments in which BM Tregs were exchanged at a higher frequency than Tregs from other tissues. This indicates that they are highly migratory, but preferentially return to the marrow. Additionally, we found that BM Tregs are high producers of IL-10, an immunosuppressive cytokine that limits inflammation. Indeed, Tregs are the primary source of this cytokine in the marrow. Loss of IL-10 production by Tregs altered the BM microenvironment in several ways. Numbers of CD51+ stromal cells, and PDGFRα+ bone forming progenitors declined. There was a marked shift towards myeloid cell production, a decrease in B cell production, and perturbed differentiation of stem and progenitor cells in IL-10 deficient marrow. Remarkably, Tregs enhanced the supportive capacity of stromal cells through IL-10 by promoting the maintenance of hematopoietic stem cells. Collectively, our observations suggest that Tregs residing within BM are master regulators of hematopoiesis and that production of IL-10 accounts for at least some of their functions. This and other properties have the potential to be exploited in the context of transplantation or hematopoietic disease.
Combined immune and airway epithelial cell profiles define asthma severity phenotypes
Authors: Oriss Timothy B , Zhou Xiaoying , Camiolo Matthew , Scholl Kathryn L , Gorry Michael , Gauthier Marc , Ray Prabir , Wenzel Sally E , Nadeau Kari Christine , Ray Anuradha 
Published: January 2023

Abstract
Asthma constitutes a spectrum of conditions that affect a substantial proportion of the population in developed countries. A large segment of asthmatics are characterized by an immunologic type 2 allergic profile. A preponderance of conventional and biologically-based therapies are available to treat these subjects. However, a small minority of individuals clinically classified as severe asthmatics (SA), are highly refractory to treatment and account for a significantly disproportionate share of the clinical and economic impact of the asthma condition as a whole. In order to better define the immune response in SA with the ultimate goal of developing novel therapeutic modalities, we analyzed bronchoalveolar lavage (BAL) cells from a cohort of SA, mild to moderate asthmatics (M/M), and healthy controls by mass cytometry (CyTOF). Unbiased bioinformatic assessment revealed 34 clusters of BAL cells with assignment of subjects to one of four groupings by principal component analysis (PCA). The majority of SA (78.9%) fell into two of these groups and constituted 65.2% of their makeup, with M/M representing the remaining 34.8%. Similarly, analysis of airway bronchial epithelial cells (BECs) by RNA sequencing (RNASeq) revealed three subject groups with SA falling largely into one of these. Synthesis of CyTOF and RNASeq data in the context of relevant clinical parameters resulted in a combined profile for each subject, which was able to distinguish those with the most severe disease. Our approach demonstrates that an unbiased, multifactorial assessment of immune and epithelial profiles can predict asthma severity, but may also reveal mechanism-based clues for targeted, personalized therapeutic intervention in severe asthmatics.
The transcriptional repressor Blimp-1 promotes Th2 development and lung inflammatory disease in a house dust mite induced model of allergic asthma
Authors: Poholek Amanda C , Hettinga Angela , He Kun , Kale Sagar , Schmitz Elizabeth , Pandya Shivani , Xie Markus Ming , Dent Alexander , Oriss Timothy B , Ray Anuradha 
Published: January 2023

Abstract
Asthma is a T cell-mediated chronic inflammatory lung disease that is thought of as a type 2/Th2 disease, yet precisely how Th2 cell development in the lung is initiated is unknown. Blimp-1 is a transcriptional repressor that regulates effector T cell responses to constrain T cell-mediated autoimmunity. We describe Blimp-1 as an unexpected regulator of allergen-induced airway inflammation that is critical to promote Th2 cell development in the lung and subsequent airway inflammation. Using a house dust mite (HDM) antigen model to drive allergic lung disease, we show that the T cell specific absence of Blimp-1 leads to reduced lung inflammation, a significant decrease in eosinophil recruitment to the lung tissue, and a near absence of Th2 cells in the lung. Intriguingly, Th1 and Th17 responses are intact, suggesting Blimp-1 is specifically required for Th2 differentiation in this model. Furthermore, despite a lack of Th2 cells in the lung, IgE responses are intact, dissociating IL-4-mediated antibody driven responses from Th2 cells in the lung. Bcl6 expression is increased in both the draining lymph node and lung and using T cell specific Bcl6 and Blimp-1 double deficient mice, we find that Th2 cells are restored when Bcl6 is deleted in addition to Blimp-1. Bcl6 is a potent repressor of GATA3, suggesting Blimp-1 acts to indirectly promote GATA3 expression by suppression of Bcl6. Furthermore, we show that STAT3 signaling in T cells is responsible to upregulate Blimp-1 and subsequent Th2 development, implicating this pathway as a possible therapeutic target. Taken together, our data suggests Blimp-1 plays an unexpected and context-dependent role in allergic lung inflammation to promote Th2 cell differentiation and subsequent lung disease.
Inhibition of allergen-specific tissue resident memory Th2 cells through blockade of CD3-mediated signaling ameliorates allergic lung inflammation
Authors: Gracias Donald Tom , Gupta Rinkesh Kumar , Croft Michael 
Published: January 2023

Abstract
Allergic asthma remains a major public health concern, with more than 26 million Americans suffering from the disease. The secretion of cytokines such as IL-4, IL-5, and IL-13 and their ability to persist in the host long after the primary allergen encounter are a reason memory Th2 cells are a major driving force behind allergic asthma morbidity. The selective removal of pre-existing memory Th2 cells could be a key therapeutic venture to enhance tolerance. One such method could involve targeting CD3 through the use of inhibitory anti-CD3 antibodies which successfully induced tolerance in animal models of autoimmune diseases and also show promise in clinical trials. To determine if we could induce airway tolerance, we tested blocking CD3 complex signaling in a HDM model of allergic inflammation. Importantly, pre-sensitized C57BL/6J mice displayed strongly ablated lung inflammation and Th2 responses when treated at the time of memory T cell reactivation with F(ab′)2 anti-CD3 antibody. This treatment was found to inhibit HDM-specific CD4+ T cell proliferation and Th2 cytokine production. Interestingly, while mice treated with anti-CD3 also showed an increased frequency of Treg cells, protection to allergic inflammation was maintained even after Treg depletion. Anti-CD3 treatment was found to specifically inhibit eosinophilic lung inflammation driven by allergen-specific lung tissue resident memory (Trm) Th2 cells as determined by treatment of mice with fingolimod (FTY720), which prevents the recruitment and contribution of circulating memory T cells to the allergic response. These data suggest that blocking CD3 complex signaling could be employed to inhibit Trm recall responses to complex allergens thus promoting airway tolerance.
Endogenous UDP-Glc acts through the purinergic receptor P2RY14 to exacerbate eosinophilia and airway hyperresponsiveness in a protease model of allergic asthma
Authors: Karcz Tadeusz , Whitehead Gregory , Nakano Hideki , Jacobson Kenneth A , Cook Donald N 
Published: January 2023

Abstract
Human asthma represents a wide spectrum of pathologies that likely arise through distinct molecular pathways, and a major therapeutic goal is to target individual therapies to specific forms, or endotypes, of this disease. Here we studied the role of UDP-glucose (UDP-Glc), an alarmin which we found to be released into the airways of allergen-sensitized mice upon their subsequent challenge with allergen. UDP-Glc signals through the purinergic receptor, P2Y14R, and mice lacking this receptor display marked attenuation of both eosinophilia and airway hyperresponsiveness (AHR) in a protease model of allergic asthma. By contrast, these features of asthma were similar in wild type and P2ry14-deficient mice in a TLR-ligand model of asthma, indicating an endotype-specific role for UDP-Glc and P2Y14R. Importantly, administrationof PPTN, a small molecule antagonist of P2Y14R, also led to reduction of airway inflammation and AHR in the protease mediated asthma. Although its mechanism of action remains unclear, P2ry14 was most highly expressed in recruited eosinophils and neutrophils, suggesting UDP-Glc acts directly on these cells. Our current findings suggest that antagonists of P2Y14R might be selectively used to control asthma exacerbations in specific forms of human asthma.
Development of conventional, but not tissue resident NK cells requires pH-controlling vacuolar a2v-ATPase
Authors: Beaman Kenneth D , Schnederman Sylvia , Katara Gajendra , Dambaeva Svetlana 
Published: January 2023

Abstract
Innate lymphoid cells are abundant in uterine endometrium and their number dramatically increases during pregnancy. In addition to conventional NK (cNK) cells, unique uterine-specific NK cells populate the uterus. These, tissue-resident NK (trNK) cells are characterized by CD49a expression. A2v-ATPase is a proton pump molecule that acidifies intracellular vesicles and is critical in processing of Notch receptors. A knockout of a2v- ATPase in VAV-1 expressing cells (cells of hematopoietic origin) leads to dramatic reduction of NK cells in blood and spleen of the genetically modified animal (a2vKO mouse). Uterine tissues of a2VKO mice were analyzed for NK cell presence based on their reactivity with the lectin DBA. DBA-reactive cells were readily observed in uterine stroma of pregnant a2vKO mice. Although the number of DBA+ cells in a2vKO mice was lower than in wild type (WT) controls, the difference was not statistically significant. Flow cytometry analysis of uterine tissues from virgin a2vKO mice revealed a dramatic decrease of cNK cells (NK1.1+NKp46+CD11b−/lowDx5+CD49a−). However, the frequency of trNK cells, that possess phenotype NK1.1+NKp46+CD11b−Dx5−/+CD49a+, was comparable with what was found in WT. Similar findings were observed in pregnant a2vKO mice. Despite the absence of cNK cells that compose a significant proportion of lymphocytes in pregnant uterine tissue, mice fertility was not critically affected and this suggested a role for trNK cells in supporting pregnancy. In conclusion, the development of NK cells that takes place in endometrium appears to be different from NK cells originated in bone marrow, namely it is not dependent on a2vATPase; this can explain uniqueness of trNK cells.
Intrinsic factor Recognition Promotes Th17/Th1 Autoimmune Gastric Inflammation in Patients with Pernicious Anemia
Authors: Troilo Arianna , Grassi Alessia , D'Elios Mario Milco 
Published: January 2023

Abstract
The intrinsic factor (IF) is the major humoral autoantigen in pernicious anemia (PA)/autoimmune gastritis. Although many studies have examined the autoantibody response to intrinsic factor and H+,K+-ATPase, no information is available on possible pathogenic mechanisms mediated by IF-specific gastric T cells. The aim of this study was to investigate IF-specific T cells in the gastric mucosa of PA patients and define their functional properties. In vivo-activated T cells from the infiltrates of the gastric mucosa of PA patients were isolated and cloned. We investigated the ability of the gastric T-cell clones to proliferate and produce cytokines in response to IF, as well as their helper function for antibody production and their cytolytic potential. For the first time we provide evidence that gastric mucosa of PA patients harbour a high proportion (20%) of autoreactive activated CD4+ T-cell clones that specifically recognize IF. Most of these clones (94%) showed a Th17 or Th1 profile. Virtually all IF-specific clones produced TNFα, IL-21 and provided substantial help for B-cell immunoglobulin production. Most mucosa-derived IF-specific T-cell clones expressed perforin-mediated cytotoxicity and induced Fas-Fas ligand-mediated apoptosis in target cells. Altogether, our results suggest that chronic autoantigen-induced T cell– dependent B-cell activation is responsible for the local synthesis of IF autoantibodies found in the sera and indicate that activation of IF-specific Th17 and Th1 T cells in the gastric mucosa represent a key effector mechanism in PA suggesting that the Th17/Th1 pathway may represent a novel target for the prevention and treatment of the disease.