bluuebunny/crossref_metadata_2025_split huggingface dataset:
A hypomorphic Ephb2 gene variant contributes to autoimmune diabetes in NOD mice associated with altered Mapk3 activities in B-lymphocytes
Authors: Wang Qiming , Chapman Harold D , Racine Jeremy J , Serreze David 
Published: January 2023

Abstract
β-cell destruction in type 1 diabetes (T1D) is considered a T-cell mediated autoimmune disease, but based on studies in the NOD mouse model, B-lymphocytes also play a critical pathogenic role. They do so by serving as a preferential subset of pathogenic APC that expand diabetogenic T cells. Immunological tolerance induction mechanisms that normally eliminate or inactivate autoreactive B-lymphocytes are defective in NOD mice and also in T1D patients. We previously mapped a faulty NOD B-lymphocyte tolerance phenotype to a 6.22Mb region on Chromosome 4. With a combination of differential gene expression and ingenuity pathway analyses (IPA) we identified a hypomorphic Ephb2 allelic variant within this region on Chromosome 4 as a potential candidate gene interacting with human T1D susceptibility genes in a common pathway modulating Mapk3 activities. Ephrin-B1 ligand engagement of Ephb2 attenuated B-cell receptor (BCR) induced Mapk3 phosphorylation in vitro. We tested the candidacy of Ephb2 by introducing into NOD mice a transgenic higher expression variant of Ephb2 derived from the B6 strain (NOD-Ephb2B6). Elevated expression of Ephb2 afforded significant T1D protection due to changes in hematopoietic cells. Despite having enhanced proliferative capacity, B-lymphocytes from NOD-Ephb2B6 mice had diminished diabetogenic activity associated with attenuated Mek-Erk signaling. Introduction of the Ephb2 transgene into NOD BCR transgenic models expressing autoreactive peripherin (Per) specific Ig molecules also inhibited the accelerated T1D development characterizing these stocks. Thus, signaling via Ephb2 in B-lymphocytes can fine-tune BCR signaling that ultimately determine their diabetogenic activities.
Monocyte activation in cardiovascular disease through inflammatory signaling associated with CD11c function
Authors: Hernandez Alfredo Alexander , Foster Greg A , Dai Yao , Rivara Kathryn Rose , Bailey Keith , Singh Gagan D. , Simon Scott I 
Published: January 2023

Abstract
Circulating monocytes are critical in the development and progression of atherosclerotic disease. During disease, prolonged metabolic dysregulation and low-grade inflammation promote monocytosis and subsequent skewing of the circulating monocyte repertoire to favor an expanded frequency of intermediate monocytes (CD14+CD16+CCR2, iMo). Intermediate monocytes display a hallmark upregulation of β2-integrin CD11c/CD18 which works in cooperation with β1-integrin VLA-4 to mediate monocyte capture and firm arrest to endothelial VCAM-1 at sites of disturbed flow at sites of nascent plaque. Notably, the expanded frequency, quantity of iMo CD11c receptors, and CD11c/VLA-4 dependent arrest of iMo to VCAM-1 gauge the severity of cardiovascular disease and are predictive of poor clinical outcome. Using these observations, we sought to examine the functional activity of arrested iMo on a substrate of VCAM-1 under arterial shear stress through the assessment of CD11c outside-in signaling using microfluidic platforms. We find that iMo undergo rapid transmigration (~90sec) at near 100% efficiency following by characteristic ADAM17 dependent shedding of CD16 that coincided with nuclear translocation of NFκβ, and elaboration of intracellular MMP-9 and IL-1β inflammatory molecules. Phenotypic conversion towards activated M1-like macrophages was governed by an allosteric shift in CD11c from high to low affinity on a majority of adherent iMo from CAD and NSTEMI patients, but not those from age-matched healthy subjects. We conclude that CD11c is a molecular biomarker and functionally relevant inflammatory signaling molecule on iMo. Its expression and function provides the most sensitive measure for diagnosing progression of CAD to MI.
CNS HIV virus persistence irrespective of CD4 T cell trafficking
Authors: Kohlmeier Alison , Lipscomb Jonathan , Li Jin-fen , Tino Ariana Santos , Anderson Albert M , Johnson Jeffrey A 
Published: January 2023

Abstract
Incomplete HIV virologic suppression in the central nervous system (CNS) can cause persisting neurologic disorders even with suppressed plasma viremia. To better understand immune properties that support CNS virus persistence we characterized T cells and virus genotypes from cerebrospinal fluid (CSF) of HIV-infected participants to understand local cell trafficking behavior and the influence of combination antiretroviral therapy (ART). Compared to blood, CSF T cells expressed higher frequencies of CCR5****, CD69****, CX3CR1****, CD39**, and HLADR**(P values:*≤0.05, **≤0.01, ***≤0.001,****≤0.0001). Among CSF T cells, CD4 T cells were more likely to express CCR7****, CX3CR1****, and CD39**, and less likely to express CCR5*. CX3CR1 expression on CSF CD4 T cells correlated with CSF viral load (VL)*, whereas, CD39 expression mirrored CSF** and plasma VL*. In the absence of ART, CSF HIV (VL&gt;900 copies/mL) shared majority sequence similarity to plasma virus demonstrating a migratory influence of susceptible cells into the CNS. However, CSF CD4 T cells from persons on ART expressed decreased CCR5**, CX3CR1****, HLADR****, and CD39***, regardless of viral suppression, while matched-PBMC CD4 T cells did not exhibit these trends. CSF CD4 T cells expressed features consistent with immune-restricted localization but with a migratory profile indicative of immune surveillance function and lymphoid trafficking. ART use associated with reduced migratory properties regardless of CNS viremia, suggesting ongoing CNS HIV replication under therapy involves local susceptible cells distinct from circulation. This CSF cellular profile could serve as a reference to target biomedical interventions that improve HIV suppression in the CNS.
Human interleukin-2 receptor beta mutations associated with defects in immunity and peripheral tolerance
Authors: Zhang Zinan , Gothe Florian , Pennamen Perrine , James John , McDonald David , Mata Carlos , Modis Yorgo , Alazami Anas , Acres Megan , Haller Wolfram , Bowen Claire , Doffinger Rainer , Sinclair Jan , Brothers Shannon , Zhang Yu , Matthews Helen , Naudion Sophie , Pelluard Fanny , Alajlan Huda , Yamazaki Yasuhiro , Notarangelo Luigi , Thaventhiran James , Engelhardt Karin , Al-Mousa Hamoud , Hambleton Sophie , Rooryck Caroline , Smith Kenneth G.C. , Lenardo Michael J 
Published: January 2023

Abstract
Interleukin-2, which conveys essential signals for effective immunity and immunological tolerance, operates through a heterotrimeric receptor. Genetic deficiency of the alpha or gamma chain causes debilitating disease. Here we identify human interleukin-2 receptor (IL-2R) beta chain (IL2RB) gene defects as a cause of life-threatening immune dysregulation. We report three homozygous mutations in the human IL2RB gene of eight individuals from four consanguineous families that cause disease by distinct mechanisms. Nearly all patients presented with autoantibodies, hypergammaglobulinemia, bowel inflammation, dermatological abnormalities, and cytomegalovirus disease. Patient T lymphocytes lacked surface expression of IL-2Rbeta and were unable to respond normally to high-dose IL-2 stimulation. By contrast, patient natural killer (NK) cells retained partial IL-2Rbeta expression and cytotoxic function. IL-2Rbeta loss of function was recapitulated in a recombinant system, in which endoplasmic reticulum sequestration was revealed as the mechanism by which certain missense mutations cause disease. Hematopoietic stem cell transplant resulted in resolution of clinical symptoms in one patient. The hypomorphic nature of this disease highlights the significance of variable IL-2Rbeta expression between different lymphocyte subsets and during their maturation as a means of modulating immune function. Insights from these patients can inform the development of IL-2-based therapeutics for immunological diseases and cancer.
Development of a high throughput, bead-based flow cytometry panel for T-helper cell cytokine quantitation
Authors: O’Rourke John , Weldon Caroline , Keenan Lisa , Carter mark B 
Published: January 2023

Abstract
Upon T-cell activation, naïve CD4+ cells differentiate into various types of T-helper (Th) cells, which is influenced by the cytokine milieu present during antigen engagement and is further driven by autocrine mechanisms. Th subsets orchestrate adaptive immune responses to specific pathogens and dysregulation of Th cells are major contributors to autoimmune and allergic diseases. Th subsets can be identified, in part, by their unique cytokine secretion profile.
To measure cytokine secretion from Th cells, Intellicyt has developed multiplex bead-based panels (QPanels) to quantitate Th1 and Th2 (4-, 6- or 9-plex) and Th1, Th2, & Th17 (7-plex) associated cytokines in cell supernatants. These miniaturized assays require sample volumes of only 10 μl. Data acquisition is performed using 96 or 384 well plates on the iQue Screener platforms. Pre-mixed reagents and cytokine standards, and templated data analysis, including event gates, gating strategy, 4 parameter standard curves and sample quantitation are auto-generated to allow for ease of use.
For proof of concept studies, peripheral blood mononuclear cells (PBMCs) from 3 donors were cultured in media containing human sera and IL-2. Cells were stimulated with anti- CD3/CD28 beads and cytokine secretion was measured 1, 2, and 3 days post-stimulation. Donor to donor variation and temporal differences in secreted cytokine levels were observed. All donor PBMCs secreted IFNγ, GM-CSF and TNFα by 3 days post-stimulation. Notably, one donor showed a more robust Th2 (IL-4, IL-6, and IL-10) and Th17 (IL-17A) response compared to other donors. These data highlight the use of QPanels in T cell screening, which allows easy assessment of effector function and the identification of Th subsets.
Function of Protein Kinase CK2 in Innate Immune Cells in Neuroinflammation
Authors: Qin Hongwei , Yang Wei , Yan Zhaoqi , Benveniste Etty 
Published: January 2023

Abstract
Immune dysregulation is critical to the pathogenesis of autoimmune diseases, including multiple sclerosis (MS). Studies in MS patients and an animal model of MS, experimental autoimmune encephalomyelitis (EAE), have highlighted the importance of both innate and adaptive immune responses in MS/EAE pathology. Antigen-presenting cells of the innate immune system, especially dendritic cells (DCs), not only activate naive CD4+ T cells by antigen presentation, but also control T cell differentiation by expressing co-stimulatory molecules and cytokines. Our current studies demonstrate that Tamoxifen-induced global deletion of CK2a in CK2αfl/flERT2Cre mice results in delayed onset and significantly slower progression of EAE, which is not observed in CK2α T cell-specific deleted CK2αfl/fldLckCre mice, which exhibit lower EAE clinical scores during the peak and resolution phases of disease. EAE mice with global CK2α deletion express reduced pathogenic Th1 and Th17 cells, and enhanced anti-inflammatory Treg cells. We speculate that CK2 kinase activity in DCs is indispensable for the initiation and progression of EAE. Specifically, our data indicate that deletion of CK2α reduces the activation of bone marrow-derived DCs, but not macrophages, by suppressing the expression of CD40, TNF-α, iNOS and IL-12 and activation of AKT and p38. Furthermore, we found that activation of DCs is prevented in EAE mice with global CK2α deletion, which correlates with differential regulation of Th1, Th17 and Treg cells. Collectively, our findings establish a novel regulatory role for CK2 kinase in DCs during EAE initiation and progression, which may provide evidence to establish CK2 as a potential novel therapeutic target for MS treatment.
Perturbation of Developmental Hematopoiesis Shapes Lung ILC2 Immune Memory
Authors: Lopez Diego A , Beaudin Anna E. 
Published: January 2023

Abstract
Our lab has previously identified a developmentally-restricted hematopoietic stem cell (drHSC) that only exists in the perinatal period and specifically gives rise to innate-like lymphocytes. The identification of a developmentally-limited cell of origin for innate-like immune cells that persist across the lifespan defines a “critical window” for immune development, in which the phenotype of the adult immune system can be shaped by extrinsic inputs. To test how developmental perturbation during this critical window drives immune dysfunction, we examined underlying changes to innate-like immune cells in the lung and susceptibility to airway dysfunction following maternal immune stimulation. Lung type-2 innate lymphoid cells (ILC2s) are recognized as potent producers of IL5 and IL13, cytokines important for eosinophil recruitment, activation and goblet cell hyperplasia during allergic airway inflammation. Surprisingly, maternal immune stimulation via a single low-dose injection of poly (i:c) at mid-gestation robustly increased the cellularity of lung ILC2s in offspring at postnatal day 14, concomitant with drHSC and ILC2 progenitor expansion during fetal development. Additionally, lung ILC2s exhibited heightened IL5 and IL13 production upon in-vitro stimulation with PMA and ionomycin in poly (i:c) perturbed offspring. Ongoing experiments examine how altered ILC2 functionality, as a result of developmental perturbation, underlie susceptibility to allergic asthma in response to secondary house-dust mite immune stimulation. Together, our data suggest perinatal immune perturbation may contribute to allergic asthma susceptibility by altering drHSC establishment and training lung ILC2 “innate immune memory”.
m6A RNA modification controls transitions between quiescence and proliferation during B cell development
Authors: Zheng Zhong , Hsu Phillip J , Cui Xiao-Long , Tan Haiyan , Zhang Linda , Sun Hui-lung , Zhang Michelle , Peng Junmin , He Chuan , Huang Haochu 
Published: January 2023

Abstract
N6-methyladenosine (m6A) is the most abundant internal modification in eukaryotic mRNA. Recent conceptual and technological advances have sparked extensive research interests in the functional roles of m6A. The writers, erasers and readers of m6A have been identified and characterized in a number of biological processes in recent years. However, the physiological roles of m6A and its effectors in lymphogenesis remained unknown. Here, we report that loss of METTL14, a core component of the m6A writer complex, in mouse B lineage cells blocked B cell development. Deletion of METTL14 in developing B cells caused dramatic reductions in the mRNA m6A level, together with an incomplete blockage during the transition from the quiescent pro-B stage to the proliferating large pre-B stage as well as a complete blockage during the proliferation-to-quiescence transition between large pre-B and small pre-B stages. We found that &gt;67% transcripts contain m6A in developing B cells, suggesting that m6A might be widely involved in gene expression regulation. m6A controlled the quiescence-to-proliferation switch partially through its cytoplasmic reader YTHDF2, as deleting YTHDF2 partially mimicked the earlier incomplete blockage between pro-B and large pre-B stages due to METTL14 deficiency, but did not cause the later proliferation-to-quiescence blockage. Altogether, our data identifies m6A, which specifically controls transitions between quiescence and proliferation, as a critical new layer of gene expression regulation in B cell development.
Selective inhibition of gelatinases modulates cellular energetics in CD4+ T-cells
Authors: Onwuha-Ekpete Lillian C. , Tokmina-Roszyk Dorota , Fields Gregg B. 
Published: January 2023

Abstract
The gelatinases, MMP-2 and MMP-9, are proteolytic enzymes with diverse physiological roles. They are linked with various aspects of the immune response, such as leukocyte trafficking and cytokine activation. However, their dysregulation is associated with various pathologies such as metastasis in cancer progression and blood-brain barrier disruption in multiple sclerosis. Much of what is known about the gelatinases is affiliated with their activities in the extracellular milieu. However, recent studies have suggested that they contribute to normal intracellular function of various immune cell types, including T-cells. CD4+ T-cells play a key role in linking the innate and adaptive arms of the immune response, as well as determining the character of the immune response.
Our findings show, for the first time, that intracellular gelatinase activity can modulate T-cell bioenergetics, affecting both oxidative phosphorylation and glycolytic pathways and resulting in a mitigated CD4+ T-cell response to stimulation. Upon stimulation, CD4+ T-cells lacking gelatinase activity demonstrate a reduced capacity to function as determined by the expression of early activation markers, proliferation, and the production of effector cytokines. In addition, RNA-seq analysis of control and gelatinase inhibited CD4+ T-cells highlight key proteins associated with T-cell receptor signaling, cytokine production, and metabolism. These results suggest that gelatinases can be immunomodulatory agents in CD4+ T-cell mediated diseases, such as multiple sclerosis, and thus have therapeutic potential.
T-cell microvilli constitute immunological synaptosomes that are specialized for intercellular communications
Authors: Jun Chang-Duk , Kim Hye-Ran 
Published: January 2023

Abstract
Lymphocytes contain abundant flexible projections termed microvilli, the inside of which contain many parallel bundles of actin filaments that extend the cell membrane in the form of a finger. Previous reports demonstrated microvilli on T cells have been proposed to survey surfaces of antigen-presenting cells (APC) or facilitate adhesion under flow. However, in addition to their key role in antigen sensing on APCs, whether they serve essential functions during T cell activation remains largely unclear. In this study, we observed that single T cell contacts with APCs occur through microvillar extensions, which appear to serve as locations for sequestration of immunologically important molecules, including TCR complexes, costimulatory and adhesion molecules, and various cytokines. Strikingly, we found that microvilli were separated from the T cell body by the combined action of two independent mechanisms (trogocytosis and membrane budding) and are deposited at the surface of cognate APCs, thereby potentially acting as the most efficient and effective means of delivering T cell messages to cognate APCs. Consistent with this potential role, these T cell microvilli-derived particles (TMPs) were independently capable of activating cognate dendritic cells (DCs). Therefore, our findings suggest that T cell microvilli might serve as ‘immunological synaptosomes’ with TMPs as a novel class of membrane vesicles serving as conveyors of T cell messages or traits to cognate APCs.
Regulatory T cells perform antigen specific suppression by depleting cognate peptide-MHC class II via trogocytosis
Authors: Akkaya Billur , Oya Yoshihiro , Akkaya Munir , Al Souz Jafar A , Holstein Amanda H , Kabat Juraj , Kamenyeva Olena , Dorward Dave , Glass Deborah , Shevach Ethan M 
Published: January 2023

Abstract
Tregs are professional suppressors of the immune response, yet their mechanism of action in vivo remains unclear. Using electron microscopy and dynamic intravital two-photon microscopy, we revealed that unlike effector T cells, Tregs displayed a distinct morphology with membrane projections that resemble filopodia at the DC binding site increasing the surface area of the contact dramatically. Despite the intense contact with DC, Tregs only abolished the interactions of effector T cells with DC when the effectors shared the same antigen specificity with Tregs. However, this cessation of the effector T cell interactions was not due to a physical hindrance created by the superior binding ability of Treg since the antigen specific inhibition of T cell-DC interaction was still observed after Tregs are removed from the environment. Instead, the antigen-specific inhibition was secondary to the removal of cognate peptide-MHC Class II (pMHCII) from the surface of DC as evidenced by the strictly antigen-specific depletion of the pMHCII from double antigen pulsed DC. pMHCII capture was only performed by antigen specific Treg, not by polyclonal Treg and it was not secondary to the interaction of CTLA-4 with B7 molecules suggesting the functional importance of TCR in Treg suppression. Prominent members of the B7 family, that are not de novo expressed in Tregs, such as CD86, ICOSL and PD-L2 were acquired alongside the TCR, suggestive of a process that includes a membrane transfer event called trogocytosis. Altogether, we propose antigen specific depletion of peptide-MHCII complexes via trogocytosis as a new mechanism for Treg-mediated suppression.
Human natural regulatory T cells recognize peptides of the heavy constant region of immunoglobulins presented by IgG+ B cells
Authors: Hsieh Li-En , Sidney John , Behnamfar Negar , Burns Jane C. , Boyle David , Firestein Gary S. , Sette Alessandro , Franco Alessandra 
Published: January 2023

Abstract
Human natural regulatory T cells (nTreg) recognize conserved peptides derived from the heavy constant region of IgG (Fc). The fine specificity of nTreg and the immunodominance of peptides was determined in healthy donors and rheumatoid arthritis (RA) patients. Eight Fc peptides were identified as highly immunogenic in all subjects with similar ranking and four of eight bound multiple class II HLA alleles. These Fc peptides are presented by IgG+ B cells by internalization and processing of surface IgG and are recognized by Fc-specific nTreg from both healthy donors and RA patients. However, exogenous IgG processed by antigen presenting cells (APC) led to the expansion of this nTreg population only in healthy donors. While nTreg from patients with RA responded to Fc peptides, the response to exogenous IgG presented by APC was compromised. These results demonstrate a hole in the central adaptive immune regulation in RA.
HIF-1α is essential for T cell suppression by murine LP-BM5 retrovirus infection-augmented Monocytic Myeloid Derived Suppressor Cells
Authors: Green Kathy A , Cramer Robert A , Green William R 
Published: January 2023

Abstract
Compared to naïve mice, monocytic myeloid-derived suppressor cells (M-MDSCs) increase in number, and suppressive efficiency, after infection with the immunodeficiency causing mouse retrovirus, LP-BM5. In ex vivo suppression assays, M-MDSCs, enriched from spleens of LP-BM5 infected mice inhibited both T- and as a novel finding, B-cell responsiveness. For M-MDSC suppression of T-cell proliferation and IFN-gamma production, we previously reported the mechanism to be almost completely iNOS/NO dependent. In contrast, the suppression of B-cell proliferation and IL-10 secretion by predominantly T2-B cells was only ~50% iNOS/NO dependent, with V-domain Ig suppressor of T cell activation (VISTA), superoxide and peroxynitrite also playing substantial effector roles. In light of recent reports on the Hypoxia-inducible transcription factor 1, α subunit (HIF-1α) regulating the function and differentiation of MDSCs, and reports of its role as a principal regulator of NO production, we describe here the characterization of M-MDSCs from myeloid-specific HIF-1α deficient, vs. w.t., B6 mice. As early as 5 weeks post infection, enriched M-MDSC preparations from HIF-1α deficient mice were significantly impaired functionally, in some cases with a total inability, to suppress in vitro T cell responsiveness. However, these same M-MDSC preparations suppressed B-cell responsiveness at a level very similar to that of control w.t. M-MDSCs. Experiments are in progress to further understand the role HIF-1α plays in the impairment of M-MDSC ex vivo suppression of T cell proliferation and function, including a possibility that HIF-1α regulates iNOS/NO production in M-MDSCs only when in the process of suppressing T- but not B-cell targets.