bluuebunny/crossref_metadata_2025_split huggingface dataset:
T cell derived XCL1 facilitates the interaction with XCR1+ DCs within the CNS during WNV encephalitis
Authors: De La Torre Kristen , Durrant Douglas 
Published: January 2023

Abstract
West Nile Virus (WNV) is a mosquito-borne flavivirus that causes encephalitis, especially in elderly and immunocompromised individuals. Previous studies have demonstrated that CNS-localized dendritic cells (DCs) protect against WNV neuroinvasive disease by establishing antiviral T cell responses in the CNS. In this study, we investigated the role of the XCL1-XCR1 interaction since its primary function is to promote DC-mediated CD8+ T cell immunity. XCR1, a chemokine receptor expressed predominately by a subset of DCs, mediates their migration in response to its ligand XCL1. We found that XCR1-deficient (XCR1−/−) mice demonstrated increased symptomatic disease and mortality after subcutaneous infection with WNV. In addition, WNV-infected XCR1−/− mice exhibited significantly increased viral burden in the CNS at day 12 post-infection. Within the WT CNS, we found increased XCL1 expression on day 6 post-infection. In contrast, the expression of its receptor, XCR1, increased on day 9 post-infection compared with uninfected CNS. As XCL1 is known to be produced by NK and CD8+ T cells, we next sought to determine which cells express XCL1 and at which locations during WNV encephalitis. Interestingly, compared with splenic T cells, CD8+ T cells isolated from the CNS and cervical lymph nodes from WT mice showed high XCL1 expression. These observations lead us to hypothesize that T cell derived XCL1 facilitates interactions with XCR1+ DCs within the CNS during WNV encephalitis.
Supported by grant from NIH (SC3 GM130472-03)
IL-9 has an important role in promoting Barrett’s esophagus (BE) phenotype in eosinophilic esophagitis (EoE)
Authors: Venkateshaiah Sathisha Upparahalli , Sekhar Yadavalli Chandra , Lokanatha Oruganti , Mishra Anil 
Published: January 2023

Abstract

Objectives
Establish the role of IL-9 in development of Barrett’s esophagus (BE) in human EoE.

Methods
Histopathological, immunohistochemical, RT-PCR and ELISA analyses are performed to identify the signature genes and proteins involved in the progression of BE in EoE.

Results
We detected characteristic feature of BE like intermediate columnar type epithelial cells, induced BE signature genes like ErbB3, CDX1, ErbB2IP in the esophageal mucosa of EoE patients. Further, we observed BE associated diagnostic proteins (TFF3, p53) and the progression markers (EGFR, p16), MHC molecules (MICA, MICB) in esophageal biopsies of chronic EoE patients. Furthermore, we also detected mucin-producing columnar cells and MUC-2, MUC-4 and MUC5AC genes and proteins along with induced IL-9 in chronic EoE patients. Interestingly, a strong correlation of IL-9 with mucin genes were observed that indicate IL-9 may have an important role in BE development in chronic EoE patients. Herein, we present in vitro data of IL-9 exposure to primary esophageal epithelial cells provide evidence that IL-9 is involved in the induction of mucins producing and epithelial barrier cells transcripts and proteins like CK8/18, GATA4, SOX9, TFF1, MUC5AC, including tight junction proteins that are the markers of BE.

Conclusions
Taken together, we provide evidence that IL-9 has a role in the initiation and progression of BE characteristics like development of mucin producing columnar epithelial cells in chronic EoE patients.
Supported by grants from NIH (R01 AI080581-Anil Mishra)
Characterization of a new cytokine complex from the IL-6/IL-12 family
Authors: Rousseau Marine , Laplante Véronique , Nadeau Ulysse , Pasquin Sarah , Fajardo Ernesto , Lesage Sylvie , Gauchat Jean-François 
Published: January 2023

Abstract
Cardiotrophin-like cytokine factor 1 (CLCF1) is a neurotrophic factor initially identified as a B lymphocyte stimulating factor. Mutations in CLCF1 cause type 2 Cold-Induced Sweating Syndrome, a severe multisystem disorder coupled with recurrent childhood infections. The CLCF1-CRLF1 composite cytokine mediate its neurotrophic effects via the CNTF receptor (CNTFR). Although immune cells do not express CNTFR, immunological activities of CLCF1 have been described, suggesting the existence of an alternative receptor. Preliminary results suggest that CLCF1 can associate with the Epstein-Barr virus-induced gene 3(EBI3). EBI3 is a subunit of the composite cytokines IL-27, IL-35 and IL-39, which exhibits immunoregulatory functions. Our hypothesis is that CLCF1 associates with EBI3 to activate an alternative receptor expressed by immune cells. The aim of the project is to identify the cells producing CLCF1/EBI3, to define its receptor and to characterize the influence of CLCF1/EBI3 on lymphocyte proliferation, differentiation, and function. We find that EBI3 and CLCF1 subunits can associate extracellularly to form a CLCF1/EBI3 complex. Moreover, we uncovered that this complex can bind to IL-12Rß1, IL-23R and CNTFRα receptor chains. Altogether, our data strongly suggest that the CLCF1/EBI3 complex is likely to assemble in vivo and act as a cytokine through known receptor chains. Overall, defining the cellular targets and biological activities of CLCF1/EBI3 will pave the way for the development of treatments in autoimmune diseases involving CLCF1 and EBI3, adding to the increasing number of biologic drugs targeting cytokines, their receptors, or signaling pathways that are used clinically to treat inflammatory diseases.
Supported by IRSC (RNI00404)
HSC-independent definitive lymphopoiesis persists into adult life
Authors: Yoshimoto Momoko , Cheng Haizi , Nishida Chika , Latorre Augusto , Kobayashi Michihiro 
Published: January 2023

Abstract
The stem cell theory that all blood cells are derived from hematopoietic stem cell (HSC) is a central dogma in hematology. However, exceptions are the fetal hematopoiesis in which primitive and definitive type of erythromyeloid cells are already produced from hemogenic endothelial cells (HECs) before the first HSC emergence in the mouse embryo. This early blood cell production prior to HSC emergence is called HSC-independent hematopoiesis and considered to be transient during fetus. Recent lineage-tracing studies revealed that some tissue-resident innate immune cells (e.g. brain microglia, mast cells, and certain types of γδT cells) are also produced independently of HSCs from early extra-embryonic yolk-sac (YS). Definitive T&B-lymphoid potential and multi-potent progenitor (MPP) potential have been detected in the YS and/or embryos in in vitro studies, however, the physiological presence of these cells in the embryo at pre-HSC stage remains unknown. In addition, whether HSCs in the fetal liver are the main source of innate-like B-1a cells has been controversial. Here, using complementary lineage tracing mouse models, we show that HSC-independent MPPs and HSC-independent adoptive T & B-lymphocytes persist into adult life and are gradually replaced by HSC-derived cells. Furthermore, we found that HSCs minimally contribute to the peritoneal B-1a cells, and most B-1a cells are originated directly from ECs in the YS and embryo and HSC-independent for life. Our discovery of extensive HSC-independent MPP and adoptive lymphocytes in adults attests to the complex blood developmental dynamics through embryo to adult that underpin the immune system and challenges the paradigm of HSC theory in hematology.
Supported by grant from NIH R01AI121197.
Vhl deficiency in Dmp1-expressing cells affects myeloid development and erythropoiesis: possible effects on myeloerythroid metabolism
Authors: Chicana Betsabel J , Emery Janna , Taglinao Hanna , Donham Cristine , Manilay Jennifer 
Published: January 2023

Abstract
The specific types of bone marrow (BM) stromal cells and local BM microenvironments that contribute to the regulation of myelopoiesis are not fully understood. The von-Hippel Lindau protein (VHL) controls responses to hypoxia and could regulate immune metabolism. We investigated this in Dmp1-Cre; Vhl conditional knockout mice (cKO), where Vhl is deleted in mesenchymal stem cells, osteoblasts (OBs) and osteocytes. Similar to previous findings when Vhl is deleted in OBs, we observed low blood glucose levels in the cKO mice, which may reflect an increase in OB cellular glycolysis and increased glucose uptake in the bone. Erythropoietin (Epo) treatment in vivo reduced blood glucose levels in rats. In preliminary studies, we found elevated Epo levels in cKO peripheral blood serum and BM fluid by 6 weeks of age, and evidence for dysregulated erythropoiesis. Moreover, the cKO displayed an increased frequency of common myeloid progenitors, CD11b+ Gr1− monocytes, and CD11b+ Gr1+ granulocytes by 10 weeks of age. We hypothesize that alterations in skeletal glucose metabolism directly affect myeloerythroid development in the BM through increased Epo-receptor (EpoR) signaling. EpoR is expressed on osteoprogenitors, hematopoietic stem cells, and B cells, but whether it is expressed on myeloid progenitors is unclear. Studies are in progress to test the hypothesis that overproduction of Epo by Vhl-deficient Dmp1+ cells results in elevated EpoR signaling in myeloid progenitors and lineages; this in turn, results in an increase in myeloid cell glycolysis and Glut1 expression and proliferation. These studies are relevant for the understanding how BM microenvironmental changes can trigger adaptations in immunometabolism of myeloid cells.
Supported by grants from NIH (R15 AI154245-01, F31 AI154815), and University of California, Merced
Cellular and molecular mediators of thymic DC homeostasis and activation
Authors: Srinivasan Jayashree , Helm Bryan , Su Zhe , Yi Song , Liu Qi , Lau Ken , Ehrlich Lauren Ilyse Richie 
Published: January 2023

Abstract
Central tolerance in the thymus ensures that autoreactive thymocytes undergo negative selection or diversion to the regulatory T cell (Treg) lineage. Thymic antigen presenting cells (APCs) such as medullary thymic epithelial cells (mTECs) and dendritic cells (DCs) present myriad self-peptides to thymocytes to induce central tolerance. We and others have previously shown that thymic cDCs can be subdivided into activated CCR7+ MHC-IIhi and resting CCR7− MHC-IIlo cDC1 and cDC2 subsets with distinct functional properties, suggesting the thymic DC compartment may be more heterogeneous than previously described. Here, we performed single-cell RNA sequencing (scRNAseq) of hematopoietic APCs isolated from adult mouse thymi to comprehensively identify thymic DC subsets with distinct gene expression profiles. Our data reveal 11 transcriptionally distinct cDC subsets. We identify 6 cDC1 subsets, 3 cDC2 subsets, 2 activated CCR7+ “cDC3” subsets distinguished by expression of cDC1 and cDC2 markers, and 4 plasmacytoid DC clusters. Through functional assays, we find that the XCR1+ cDC3s are the most efficient at presenting mTEC-derived peptides on MHC-I and MHC-II, while 2 distinct cDC2 subsets are the most efficient at generating Tregs in vitro. Additional scRNA-seq and flow cytometry analyses of thymic DCs from mice in which thymocyte maturation was blocked at different developmental stages reveal candidate cellular and molecular crosstalk signals that regulate homeostasis and/or activation of pDC and cDC subsets. Our findings reveal hitherto unknown functional heterogeneity within the thymic DC compartment and identify potential interactions that support DC homeostasis and activation which in turn could impact central tolerance.
Supported by NIH P01 AI139449-02
Regulation of Thrombopoiesis by the Inhibitor of MyoD Family A (I-mfa)
Authors: Houser Jeremy S , Patel Maulin , Onopiuk Marta , Tsiokas Leonidas , Humphrey Mary Beth 
Published: January 2023

Abstract
The differentiation of megakaryocytes (Mk) from hematopoietic stem and progenitor cells (HSPC) is crucial to homeostasis and requires tight regulation. Cell-intrinsic factors skew HSPCs towards the Mk lineage and promote terminal differentiation, but this regulatory network is not yet well understood. The Inhibitor of MyoD Family A (I-mfa) is a protein which interacts with multiple pathways, including β-catenin/TCF/LEF and JNK which have known roles in HSPC differentiation. This led us to examine the role of I-mfa in hematopoiesis, hypothesizing that it promotes Mk differentiation by regulating cellular signaling events.
Mice which completely lack I-mfa expression (I-mfa KO) had reduced percentages of Mk/Erythroid Progenitors (MEPs) in the bone marrow compared to WT animals by flow cytometry, as well as reduced platelet counts in peripheral blood. To examine the potential role for I-mfa in thrombopoiesis, K562 cells (which show PMA-induced Mk differentiation) were transfected with a vector for doxycycline-induced overexpression of I-mfa, and differentiation was assessed by flow cytometry for Mk surface markers. Cells overexpressing I-mfa showed increased differentiation with a higher proportion of CD61+, CD71− cells compared to parent. The mechanism underlying this effect is being examined by analyzing cellular signaling after PMA stimulation and identifying I-mfa binding partners over time.
This work demonstrates a role for I-mfa in the regulatory control of hematopoiesis and lineage commitment through its regulation of thrombopoiesis. These results may also suggest that I-mfa could be an important cellular regulator in hematopoietic disorders including anemia, bone marrow failure, or hematological malignancies.
Supported by NIH T32 Immunology Training Grant AI00763316A1
Covalent heterobivalent inhibitor effectively inhibits anaphylaxis to peanut allergen in a humanized mouse model
Authors: Alakhras Nada Salem , Shin Jaeho , Smith Scott , Bilgicer Basar , Kaplan Mark H 
Published: January 2023

Abstract
Peanut-induced allergy is an IgE-mediated type I hypersensitivity reaction that manifests symptoms ranging from local edema to life-threatening anaphylaxis. While there are treatments for symptoms in allergic patients resulting from allergen exposure, there are few preventive therapies other than avoidance and no allergen specific therapies. We have previously designed a covalent heterobivalent inhibitor (cHBI) that binds in an allergen-specific manner as a preventive for allergic reactions. Building on previous in vitro testing, in this report we have developed a humanized mouse model to test cHBI efficacy in vivo. Humanized mice (NSG-SCF/GM-CSF/IL-3) developed mature functional human mast cells in various tissues and developed robust anaphylactic reactions when passively sensitized with human IgE monoclonal antibodies specific for peanut allergen. We found that the allergic response is IgE dose-dependent and is mediated by human mast cells indicated by elevated tryptase serum levels and the upregulation of mast cell degranulation markers. Using the validated humanized mouse model, we showed that cHBI inhibited IgE-mediated anaphylaxis and human mast cell degranulation. We demonstrated that cHBI effectively inhibits anaphylaxis for up to 14 days, and cHBI inhibition is specific to peanut allergen. Importantly, cHBI rescued the mice from lethal anaphylactic response during oral peanut-induced anaphylaxis. These findings suggest that cHBI has the potential to be an effective preventative for peanut food allergy in patients.
Comparison of sensitizing and disease-eliciting capacities of salt-soluble protein extracts from diploid (einkorn) versus tetraploid (durum) wheats
Authors: Gao Haoran , Jorgensen Rick Dean , Raghunath Rajsri , Ng Perry K W , Gangur Venugopal 
Published: January 2023

Abstract
Hypersensitivity reactions to wheat have reached significant levels of global public health concern for reasons that are incompletely understood. Einkorn wheat (Triticum monococcum, genome AA) is commercially available but is rarely consumed, whereas durum wheat (Triticum durum, genome AABB) is commonly used for pasta making. At present, most mouse models of wheat allergy are adjuvant-based, which fail to reveal the intrinsic allergenic potential of different wheat products. Here we compared einkorn and durum wheats in terms of their sensitizing and disease-eliciting capacities in an adjuvant-free mouse model of anaphylaxis induced by salt-soluble protein extract (SSPE). Balb/cJ female mice were sensitized transdermally with SSPE or saline for nine weeks. Later, systemic or oral anaphylactic reactions were elicited in these mice using SSPE via intraperitoneal or oral route, respectively. We did not observe major differences in sensitizing capacity as suggested by antibody responses (i.e., sIgE and tIgE levels) in SSPE-sensitized mice. However, SSPE from durum appears to be more potent than that from einkorn in eliciting systemic but not oral anaphylaxis as shown by hypothermic shock responses as well as mucosal mast cell degranulation. This is the first report on comparing the intrinsic allergenicity between tetraploid and diploid wheats in an adjuvant-free mouse model.
This research was funded by: the United States Department of Agriculture (USDA)/National Institute of Food and Agriculture (NIFA); Hatch project MICL02486 (Accession Number: 1012322), Hatch project MICL01699; Agricultural and Food Research Initiative Competitive Program, grant number: 2018-67017-27876; and Project GREEEN (Michigan State University); R.J., was supported by the John Harvey Kellogg Graduate Assistantship and the Academic Achievement Graduate Assistantship from the Michigan State University.
Paneth cells regulate allergic sensitization in the gastrointestinal tract
Authors: Kim Eunsoo , Burnett Drew J , Joldrichsen Marisa R , Mori-Akiyama Yuko , Cormet-Boyaka Estelle , Boyaka Prosper N 
Published: January 2023

Abstract
The gut immune homeostasis plays a major role in controlling sensitization to food allergens and the development of allergic responses in the gastrointestinal tract, but also to skin or lung. Epithelial cells sensing of commensal microbes is crucial for the maturation of the gut immune system and antibiotic treatment during early life is believed to have contributed to the increased incidence of food allergy. Paneth cells secrete antimicrobial products and cytokines in the crypts of the small intestine, but their role in food allergy remains elusive. We found that SOX9ΔIEC mice, which lack Paneth cells, display dysbiosis with increased microbial diversity. Interestingly, Paneth cell deficiency increased the of numbers of ILC2 in intestinal lymphoid tissues. Upon oral sensitization with a model food antigen in the presence of cholera toxin, SOX9ΔIEC mice developed significantly higher allergen-specific serum IgE responses, and enhanced signs of allergic responses (i.e., swelling, hypothermia, impairment of lung functions) after nasal allergen challenge. Transplantation of fecal materials from SOX9ΔIEC mice was able to transfer the hyper-allergenic phenotype to control wild-type mice. In addition, bacteria-free fecal extract from SOX9ΔIEC mice increased the number of IgE producing B cells than fecal extract from control mice with treatment of anti-CD40 and IL-4 in vitro. Ongoing studies are assessing the metabolites present in the intestinal lumen of SOX9ΔIEC mice and their effect of intestinal epithelial cells and myeloid cells to elucidate the mechanisms by which impaired or defective anti-microbial functions of Paneth cells regulate allergic sensitization of the GI tract.
Supported by Ohio State Univerisity NIH
PD-1 blockade in T follicular helper cells protects mice from peanut allergy by promoting production of low-affinity antibodies
Authors: Lama Jyoti K , Iijima Koji , Kobayashi Takao , Kita Hirohito 
Published: January 2023

Abstract
Food allergy, such as peanut allergy can cause life-threatening anaphylaxis. While T follicular helper (Tfh) cells play a major role in production of allergen-specific IgE and IgG antibodies, the immunologic mechanisms that regulate the production of antibodies that mediate anaphylaxis are not well understood. While PD-1 is highly expressed on Tfh cells, our knowledge is limited regarding the role of the molecule in Tfh cells. Thus, the objective of this study was to investigate the roles of PD-1 in allergic immune responses using a mouse model of peanut allergy. Naïve wild-type mice were exposed to peanut flour intranasally (i.n.) and then challenged by intraperitoneal (i.p.) injection of peanut extract. The mice exposed i.n. to peanut flour produced peanut-specific IgE and IgG antibodies and developed anaphylaxis when challenged i.p. with peanut extract. PD-1 blockade with anti-PD-1 treatment Tfh and germinal center B cell numbers in mice exposed to peanut flour. Nonetheless, anti-PD-1 or anti-PD-L1 fully protected mice from anaphylaxis. Anti-PD-1 or genetic deficiency of PD-1 in CD4+ T cells reduced and increased levels of peanut-specific IgE and IgG, respectively. Anti-PD-1 promoted production of low-affinity IgE and IgG antibodies with no effect on high-affinity antibodies. By passive cutaneous anaphylaxis, peanut-specific antibodies generated in anti-PD1 treated mice failed to induce anaphylaxis, and plasma from these mice prevented anaphylaxis. Therefore, blockade of the PD-1 pathway is protective against allergic immune responses. The interaction between Tfh cells and B cells may play a pivotal role in controlling antibody quality and clinical manifestation of allergic diseases.
Interleukin-10 enhances IL-33-mediated MC activation and modulates the development of food allergy
Authors: Mathias Clinton B , Krajewski Dylan , Taruselli Marcela T , Rovatti Jeffrey , Mire Mohamed , Schneider Sallie S , Ryan John J 
Published: January 2023

Abstract
We recently demonstrated an unexpected, proinflammatory role for the pleiotropic cytokine, IL-10, in promoting mast cell (MC) responses during food allergy. IL-10 enhanced MC proliferation and survival and promoted IgE-dependent responses to enteric ovalbumin (OVA) challenge. However, whether the effects of IL-10 on MCs extend beyond IgE-mediated signaling is not clear. To determine whether IL-10 can prime MC activation mediated by IgE-independent stimuli, we assessed the effects of rIL-10 on IL-33-stimulated bone marrow-derived MCs (BMMCs) and examined the development of food allergy in IL-10-depleted ST2−/− mice. IL-10 co-culture significantly enhanced the activity and cytokine production of both IgE and IL-33-activated MCs. Furthermore, the production of MC-derived cytokines such as IL-13 was suppressed in IL-33-stimulated IL-10−/− BMMCs, which could be restored by addition of exogenous IL-10. IL-10 enhanced MC FcɛRI and ST2 expression. Further, ST2−/− BMMCs demonstrated that IL-10 enhanced IgE-mediated activation in the absence of autocrine IL-33 signaling. To assess the role of IL-10 in vivo, food allergy was measured in WT and ST2−/− mice subjected to antibody-mediated IL-10 depletion. IL-10-depleted WT mice exhibited a significant attenuation in allergic diarrhea and MC-mediated responses to OVA challenge. Similarly, ST2−/− mice also exhibited a profound suppression of MC-mediated responses, demonstrating the importance of IL-33 in MC regulation. IL-10 depletion had no additional effects in ST2−/− mice. However, IL-10/ST2−/− mice exhibited a further decrease in OVA-IgE and MC activation compared to ST2−/− mice. Our data reveal a novel role for IL-10 in enhancing both IgE and IL-33-mediated MC responses.
This project was supported by funds from the National Institutes of Health grants: NIAID R15AI107668 (CBM)