bluuebunny/crossref_metadata_2025_split huggingface dataset:
Airway epithelial cell-specific deletion of EGFR in Scnn1b transgenic mice modulates inflammatory responses but doesn’t affect muco-obstructive features of lung disease
Authors: Choudhary Ishita , Paudel Kshitiz , Kumar Rahul , Patial Sonika , Saini Yogesh 
Published: November 2024

Abstract
Epidermal growth factor receptor (EGFR) is known to regulate airway mucous cell metaplasia, but the roles of EGFR-mediated pathway in the manifestation of cystic fibrosis (CF)-relevant lung phenotypes remain unclear. We hypothesized that airway epithelial cell-specific deficiency of EGFR mitigates muco-obstructive and inflammatory responses in sodium channel, non-voltage gated 1, beta subunit (Scnn1b) overexpressing -transgenic (Tg+) mice, a mouse model of human CF-like lung disease. To test our hypothesis, we generated airway epithelial cell-specific EGFR-deficient mice and performed various endpoint analyses. The deletion of EGFR in airway epithelial cells in Tg+ mice resulted in increased neutrophilic and macrophage infiltration into lung airspaces, which was accompanied by significantly increased levels of KC, G-CSF, MIP-2, MIP-1α, and MIP-1β. Additionally, as compared to EGFR-sufficient Tg+ mice, airway epithelial cell-specific EGFR-deficient Tg+ mice exhibited significantly increased postnatal mortality, compromised bacterial clearance and elevated markers of Th2 inflammation, including Il13, Slc26a4, and Retnla. Counterintuitively, the deletion of EGFR in airway epithelial cells of Tg+ mice did not alter mucous cell metaplasia, MUC5AC/5B production, and degree of mucus obstruction. Our data shows that, unlike inflammation, muco-obstruction-relevant responses in Tg+ mice are not dependent on airway epithelial cell-specific EGFR signaling.
Macrophage expressed TRIM47 is required to control RNA virus infection by targeting MAVS
Authors: Lu Wenting , xing Junji , Wang Junying , Gonzalez Anastasia , Wang Ling , Zhang Zhiqiang 
Published: November 2024

Abstract
In the past half century, there have been multiple RNA virus pandemics (influenza 1957, 1968 and 2009; SARS-CoV and COVID-19), inhabiting 44% of all emerging contagious diseases. Innate immunity is the first line of defense against viral infection. Here we found that TRIM47, a member of the TRIM family E3 ligase, plays a pivotal role in positively regulating host innate immunity in response to Influenza A virus (IAV). Among immune cells, TRIM47 is highly expressed in macrophages. Compared with wild type, TRIM47 deficient bone marrow derived macrophages exhibited a significantly lower level of type I interferon (IFN-I) following RNA mimics infection, encompassing IAV and poly I: C. After in vivo IAV infection, TRIM47 knockout mice showed an extremely lower survival rate, along with a lower level of IFN-I, higher viral load and more inflammatory cell infiltration in lungs compared to wild-type mice. Mechanistically, TRIM47 induces K68-linked ubiquitination of mitochondrial antiviral-signaling protein (MAVS), a key adaptor in RNA-sensing pathway, leading to robust IFN-I production. This K68-linked ubiquitination by TRIM47 is essential for MAVS aggregation and activation in macrophage. These findings provide compelling evidence that TRIM47 interacts with MAVS, enhancing IFN-I production to effectively restrict IAV infection in macrophages. This research not only advances our understanding of influenza pathogenesis but also opens avenues for potential therapeutic interventions.
Neuregulin-1 reduces necroptosis and protects mice from death caused by respiratory viral infection
Authors: Hussain Syed-Rehan , Rohlfing Michelle , Santoro Jennifer , Muralidharan Kaushik , Peeples Mark , Grayson Mitchell 
Published: November 2024

Abstract
Respiratory syncytial virus (RSV) and influenza type A virus (IAV) cause significant morbidity and mortality even with available vaccines. Pre-existing atopy provides protection against death in mice infected with Sendai virus (SeV), a rodent virus related to RSV. We found neuregulin-1 (NRG1) expressed at high levels in atopic mice, and when given exogenously (500 ng/day i.n for five days) to non-atopic mice provides significant protection against mortality from lethal doses of both SeV (2x106 pfu) and IAV (3x103 pfu PR8-mouse adapted). Further, using well-differentiated human airway epithelial cells, NRG-1 was found to improve epithelial barrier function following RSV infection. RSV has been reported to cause necroptosis, which can induce inflammation and alter epithelial membrane permeability. We hypothesized the survival advantage of NRG1 involves inhibition of necroptosis. Treating cells from the murine airway epithelial cell line LET-1 with NRG1 before inoculation with SeV significantly inhibited viral induced necroptosis as measured by a real-time non-lytic apoptosis/necroptosis assay. Furthermore, NRG1 reduced expression of genes promoting necroptosis (RIPK3 and MLKL) in well-differentiated human bronchial epithelial cells 72h post RSV infection. Together, these data suggest NRG1 modulates viral induced cell death through inhibition of necroptosis of epithelial cells and may explain the reduced airway leak and increased survival seen in our model.
Unfolding micro-case studies: a strategy for better student engagement/understanding/recall of difficult concepts in didactic sessions
Authors: Alard Pascale 
Published: November 2024

Abstract
The most common teaching format in professional school is lecturing, i.e., Powerpoint presentation, which minimally engages students. However, engagement of students in the classroom has been shown to improve understanding and retention of material via assessment of previous knowledge, prediction and retrieval of information. Learning from real-world scenarios is a method that medical/dental students favor over lectures covering basic science as it helps them make connections among various taught topics.
We have included very short case studies (micro-case studies) that unfold over time in 50-minute lecture sessions covering basic immunology. Basic concepts were first presented and a micro-case study developed over time to illustrate each taught concepts/details in a clinical setting. Students were asked prediction/retrieval questions about different portions of the session using a patient example. Students were provided with the micro-case study in its entirety at the end of each session, so they could use it to plug in the information learned during the session. The outcome of our study suggested that utilizing unfolding micro-case studies to illustrate basic immunology concepts is beneficial for the students in term of understanding and recalling the material better as well as enhancing student engagement as it can make challenging subjects more palatable. This format can be applied to any field and at any education level, including Graduate, Medical or Dental school.
Contribution of Helios in Ets1-Deficient NK Cells
Authors: Moos Ezra , Melanson Randy , Kee Barbara 
Published: November 2024

Abstract
Natural killer (NK) cells mediate anti-viral and anti-cancer immune responses. However, the gene networks that regulate NK cell development and function remain poorly understood. One transcription factor critical for proper NK cell development is Ets1. Previously we showed that loss of Ets1 at the germline level resulted in a developmental block, a functional defect, as well as a phenotype reminiscent of chronic cytokine stimulation in the NK cell compartment. Ets1-deficient NK cells also overexpress Helios, an Ikaros family transcription factor encoded by Ikzf2. Ikaros family transcription factors regulate development in many hematopoietic cell types; therefore, we hypothesized that Helios may be acting downstream of Ets1 to mediate the observed developmental block or functional defects. To investigate this, we generated NK cell-driven conditional deletion of Ets1 only (Ets1 cKO) or Ets1 and Ikzf2 (2xKO). The data presented here confirmed that Helios is highly expressed at the protein level in Ets1 cKO NK cells. Further, overexpression of Helios in Ets1-deficient NK cells does not contribute to the block in maturation nor the cytokine production defect. However, activated NK cells show high Helios expression, suggesting that Helios expression in Ets1-/- NK cells may be a consequence of the chronic cytokine stimulation phenotype.
Dual specificity phosphatase 12 deficiency reveals its role in maintaining T cell survival
Authors: Chen Min , Yang Chia-Yu 
Published: November 2024

Abstract
Dual specificity phosphatases (DUSPs) are protein phosphatases that regulate cellular signaling pathways by dephosphorylating threonine/serine and tyrosine residues on the substrates. DUSP12 mediates the regulation of Toll-like receptor signaling, cell cycle, hepatocyte metabolism, cardiac hypertrophy, and fibrosis. Moreover, a nonsynonymous mutation of DUSP12 has been identified in patients with T-cell-mediated autoimmune diseases. However, the regulatory roles and mechanisms of DUSP12 in T cells remain unclear. In this study, we investigated the functional regulation and molecular mechanism of DUSP12 in T cells using T cell-specific DUSP12 conditional knockout mice. We found that DUSP12 cKO mice had severe T-cell lymphopenia. More interestingly, DUSP12 cKO mice spontaneously developed systemic inflammation including splenomegaly and intestinal inflammation. Furthermore, our results showed that DUSP12 localized to mitochondria and was essential for mitochondria integrity and cellular oxidative stress in T cells. Using mass spectrometry analysis, western blotting, immunofluorescent staining, and proximity ligation assay, we identified a novel DUSP12-interacted protein in T cells. The functional regulation of DUSP12 on this novel protein in T cells was also explored. Our current results made a significant breakthrough in DUSP12 functions in T cells. Also, DUSP12 may serve as a potential biomarker and therapeutic target for inflammatory disease.
Dependence of lymphopoiesis on efficient β-catenin degradation
Authors: Cosgun Kadriye , Jumaa Huda , Robinson Mark , Klemm Lars , Oulghazi Salim , Fonseca-Arce David , Xu Liang , Xiao Gang , Khanduja Dhruv , Chan Lai , Lee Jaewoong , Kume Kohei , Song Joo , Chan Wing-Chung , Chen Jianjun , Taketo Mark , Kothari Shalin 
Published: November 2024

Abstract
Wnt/β-catenin pathway is tightly controlled by β-catenin destruction complex to regulate lineage specification and differentiation. Compared to epithelial lineage, lymphoid cells display higher rates of β-catenin degradation to achieve constitutively low levels of β-catenin. Emphasizing the reliance of lymphoid cells on β-catenin degradation, expression of degradation resistant form of β-catenin or deletion of APC or CK1 suppressed lymphoid development. Similarly, small molecules inhibiting GSK3β mediated phosphorylation or proteasomal degradation of β-catenin induced higher toxicity selectively in lymphoid cells compared to other lineages.
Our interactome and CHIP-seq analysis highlighted that the unique dependency of lymphoid cells on efficient degradation of β-catenin is mediated by Ikaros zinc finger (IKZF) proteins and nucleosome remodeling and deacetylase (NuRD) complexes. Unlike activating TCF/β-catenin complexes in epithelial cells, recruitment of repressive β-catenin/IKZF/NuRD complexes to MYC superenhancer (SE) regions lead to suppression of Myc expression and induced cell death. Deletion of both IKZF1 and IKZF3 or mutation of Ikaros binding motifs within the Myc-SE subverted the β-catenin mediated toxicity. Upon loss of Ikaros factors, β-catenin interaction with TCF-factors resulted in de novo activation of epithelial specific genes. Our findings suggest that β-catenin degradation is an evolutionary requirement for Ikaros mediated protection of lymphoid identity.
Spatial Characterization of the Immune Microenvironment within HIV+ Lymph Nodes
Authors: Furler O'Brien Robert , Dunn Daniel , Iñiguez Luis Pedro , Del Rio Estrada Perla , Avila-Rios Santiago , Nixon Douglas 
Published: November 2024

Abstract
Background: Persistent HIV replication is hypothesized to induce chronic inflammation in ART-suppressed people living with HIV (PLWH), however defining this chronic inflammation within tissues is limited. Molecular and spatial characterization of chronic inflammation within human lymph nodes from PLWH provide valuable information on the immunological microenvironment that limits viral eradication.
Methods: We spatially characterized the transcriptomic and proteomic profiles of the lymph node microenvironment from 6 human lymph nodes: three from ART-suppressed PLWH, two from chronically infected PLWH, and one from a male HIV- control donor. For spatial transcriptomics analysis, lymph nodes were either from fresh-frozen or FFPE tissue. Lymph node sections (10μm) were placed onto the 10X Visium slides, stained with H&E, and imaged prior to RNA-Seq library prep. Following sequencing cluster identification was done with single cell RNA-seq datasets and downstream bioinformatics analyses were performed.
Results: Following the bioinformatic integration of spatial transcriptomic datasets, cluster analysis identified pro-inflammatory T cells within the HIV+ tissues compared to the HIV- tissue within the T cell zone. Additionally, two sets of proinflammatory myeloid cells were identified that were spatially located within the afferent and efferent lymphatics in the HIV+ tissues. These proinflammatory clusters further characterize chronic inflammation that occurs in PLWH.
Evaluating the Functional Capability of Circulating Natural Killer Cells in Human Studies
Authors: Siksou Léa , Kong Weimin , Cohen Gadi , Kumar Gaurav , Dotan Or , Schnir Victoria , Gutman Dikla , Tamir Ayala , Rigby Mark , Graves Andrew , Orange Jordan 
Published: November 2024

Abstract
Introduction:
Interleukin-15 (IL-15) appears to be involved in T cell and natural killer (NK) cell homeostasis and immune-mediated diseases. In a first-in-human study of a monoclonal antibody against IL-15 (TEV-53408), a reduction of circulating NK cells was observed in healthy volunteers. In order to determine if there was also a qualitative effect on NK cells, in-vitro functional assessments were performed on the remaining NK cells.
Methods:
PBMCs were obtained from 81 healthy participants(administered TEV-53408 or placebo) in a single-dose (SD) and a multiple dose (MD) study. Samples were stimulated with phorbol myristate acetate (PMA)/ionomycin to induce degranulation, stained with an NK cell-extracellular panel and CD107a. After fixation, permeabilization, and intracellular staining for perforin, cells were run through a flow-cytometry assay.
Results:
The collected data demonstrated a standard CD56 bright:dim ratio at all time points. Perforin load (MFI) and % of CD107a expression with or without stimulation suggest that TEV-53408 had no effect on the degranulation ability of NK cells.
Discussion:
These assessments provide an effective method to assess the phenotype and functionality of remaining NK cells following anti-IL-15 treatment. The results suggest that treatment with TEV-53408 does not affect the functionality of the remaining NK cells during the studied timeframe, despite the reduction of circulating NK cells.
CD38 modulates CD8 TRM responses after respiratory viral infection
Authors: de Almeida Santos Gislane , Wu Yue , Sen Chaudhuri Arka , Tang Jinyi , Cheon In Su , Narasimhan Harish , Sun Jie 
Published: November 2024

Abstract
Aged hosts have increased susceptibility to develop persistent pathology and sustained inflammation after exposure to respiratory viral infection. Our group previously demonstrated that this condition is associated with an exuberant CD8 tissue resident memory (TRM) cell response. However, the underlying mechanisms that contribute to exuberant TRM accumulation and pathological activity in this context are still unknown. We found that CD8 TRM cells isolated from the lungs of influenza-infected young and aged mice express high levels of CD38. Interestingly, aging further increases the expression of this molecule on CD8 TRMs. CD38 is a molecule with NAD-depleting activity, which can affect a broad diversity of cellular processes. To address the physiological consequence of high expression of CD38 by CD8 TRMs in the lung during influenza infection, young CD38KO mice and Wild type (WT) mice were infected with influenza. We observed reduced numbers of antigen specific CD8 TRMs in the lung of infected CD38KO mice compared with infected WT mice. Mixed bone marrow chimera (1:1 ratio) experiments showed decreased frequency of antigen specific CD8 T cells in the lungs following CD38 deficiency, demonstrating an intrinsic role for CD38 in CD8 TRM responses. Similar results were observed in CD8 T cell-specific CD38-deficient mice (CD38fl/flCD8cre). Together these results show an important role for CD38 in modulating CD8 TRM responses in the lung during influenza infection.
2-Deoxy-D-glucose as a potential therapeutic agent for reducing neuroinflammation following spinal cord injury
Authors: Vinit Stephane , Michel-Flutot Pauline , Mansart Arnaud 
Published: November 2024

Abstract
High spinal cord injuries (SCIs) lead to persistent diaphragm paralysis and related respiratory dysfunctions. The absence of functional recovery is due to limited neuroplasticity processes, mainly caused by chronic neuroinflammation occurring in the injured spinal cord. In this study, we aimed at reducing glucose metabolism that supports neuroinflammatory processes in an acute preclinical model of C2 spinal cord lateral hemi-section in rats. We administered 2-Deoxy-D-glucose (2-DG; 200 mg/kg/day s.c. for 7 days) and evaluated the effect on respiratory function and chondroitin sulfate proteoglycans (CSPGs) production around spinal phrenic motoneurons. Contrary to our initial hypothesis, our 2-DG treatment did not have any effect on diaphragm activity and CSPGs production in injured rats, even if a slight increase in tidal volume can be observed. Unexpectedly, it led to deleterious effects in uninjured (sham) animals, characterized by an increased ventilation and CSPGs production. Globally, our results seem to indicate that this 2-DG treatment paradigm could create a neuroinflammatory state in healthy animals, without altering or modulating the already installed spinal inflammation in injured rats. Given the established beneficial effects of 2-DG observed in other studies on neuronal activity and inflammation, adapted2-DG doses and/or longer treatment duration should be explored for reducing deleterious inflammatory processes occurring after SCI.
Role of the cGAS DNA sensing pathway in acute intestinal ischemia-reperfusion injury
Authors: Jayakumar Preethi , Hu Jiannan , Huang Hai , Deng Meihong 
Published: November 2024

Abstract
Acute intestinal ischemia-reperfusion injury (AIRI) often occurs during trauma, shock, and surgical procedures associated with high mortality. Cyclic GMP-AMP synthase (cGAS) is a DNA sensor sensing double-stranded DNA in the cytoplasm to initiate immune responses. As circulating cell-free DNA levels are known to increase during AIRI, we hypothesized that the cGAS DNA sensing pathway is involved in the pathogenesis of AIRI. To test our hypothesis, we subjected C57BL/6 (WT) and cGAS-/- mice to 60 min intestinal ischemia via superior mesenteric artery occlusion followed by 4h reperfusion to induce AIRI. Intestinal and pulmonary injury were assessed by histology, cytokine levels, and neutrophil infiltration. The jejunum was severely injured after AIRI, evidenced by damaged intestinal villi, increased IL-6 levels, and neutrophil infiltration. The cGAS signaling was activated in the jejunum after AIRI, evidenced by increased levels of cGAS and the downstream mediator pIRF3. Notably, knockout of cGAS alleviated AIRI by attenuating the damage in intestinal villi and decreasing levels of IL-6 in the jejunum compared with WT mice. The levels of IL-6 and the numbers of neutrophils in the lung were lower in the cGAS-/- mice compared with WT mice after AIRI, suggesting that the cGAS signaling was involved in AIRI-induced lung injury. Our results demonstrate that the cGAS signaling plays critical roles in the pathogenesis of AIRI. The cGAS pathway maybe targeted for treating AIRI.
Highly efficient IL-2-dependent gene activation by a single wave of IL-2R signaling in human regulatory T cells
Authors: Vujanac Milos , Yu Aixin , Malek Thomas 
Published: November 2024

Abstract
Low-dose IL-2 is being advanced as a therapy for autoimmunity based on selectivity toward Tregs. Regarding human CD4+ Treg and effector memory (TEM) T cells, the IL-2 sensitivity of Tregs is 10- and 100-fold greater for proximal signaling and down-stream gene activation, respectively. To further investigate the efficiency of IL-2R signaling in Tregs, we monitored the persistence of a single wave of pSTAT5 and assessed if this supported IL-2-dependent transcription. When Treg and TEM cells were pulsed with a high dose of IL-2 to normalize initial STAT5 activation, pSTAT5 persisted for &gt;2 hr only in Tregs. Lower amounts of pulsed-IL-2 also led to persistent pSTAT5 in Tregs. The short persistence of pSTAT5 in TEM cells is not cell intrinsic because pulsing TEM cells with IL-7 also led to lasting pSTAT5. Thus, the high expression of CD25 and CD127 in Treg and TEM cells likely contributed to persistent pSTAT5. This conclusion implies that a component of proximal STAT5-dependent signaling is limiting in comparison to the numbers of bound receptors. RNA Prime Flow demonstrated that transient pulse of IL-2 or IL-7 in Treg and TEM cells caused upregulation of CISH mRNA. These results indicate that a limited exposure of T cells expressing high amounts of the IL-2R and IL-7R are linked to a mechanism to sustain proximal signaling to support gene transcription. This mechanism may fine tune gene expression enabling cells to maximize utilization of limiting cytokines in vivo.
Profiling the interactions of epigenetic regulators and their control of T cell states
Authors: Inge Melissa , Siggers Trevor 
Published: November 2024

Abstract
Characterizing the gene regulatory mechanisms that define distinct T cell states is essential to understanding T cell biology in health and disease. Despite advances in identifying transcriptional and epigenetic differences across T cell states, the regulators of these changes remain poorly defined. We performed a systems-level proteomic and genomic study to identify transcription factor (TF) and chromatin modifying cofactor (COF) complexes that might regulate cell state in resting, activated, and exhausted T cells. We used a high-throughput, protein-binding microarray-based approach called CoRec, to profile numerous COFs and identified unique TF-COF recruitment networks for each T cell state. In our study, some COFs are recruited specifically to TFs known to drive T cell state changes, including TFs implicated in T cell exhaustion. Exhaustion-specific recruitment of COFs suggests that these TFs may drive exhaustion through targeted alteration of epigenetic marks by these COFs. This alters expression of exhaustion-related genes and can create a functional change in T cell state. Additionally, by integrating our CoRec data with other methods, we examine how these TF-COF networks relate to chromatin accessibility and genome-wide chromatin marks. The CoRec method and its integration with other methods provides an exciting new approach for profiling cell-specific TF-COF complexes and allows us to better understand and identify the epigenetic regulators of T cell states.