bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neurokinin A reduces the influx of inflammatory cells into the skin in a mast-cell dependent manner
Authors: Barreto Joharis , Bharadwaj Shreya , Hay Ahnika , Sumpter Tina 
Published: November 2024

Abstract
Cutaneous mast cells promote immune responses but are also involved in reducing inflammation and in promoting immune resolution. Mechanisms initiating the anti-inflammatory function of mast cells are poorly defined. Mast cells reside in close proximity to neurons and their activation is modulated by neuropeptides, including the tachykinin family members. Neurokinin A (NKA) is an understudied tachykinin family member that limits cutaneous inflammation in a murine contact hypersensitivity (CHS) model but increases inflammation in colitis. In this study, the regulatory function of NKA in cutaneous inflammation is mechanistically evaluated using a robust model of DNFB-induced CHS. Administration of NKA prior to sensitization inhibited the expression of inflammatory cytokines and chemokines, including Il1b, Il6 and Ccl2. Correspondingly, NKA reduced recruitment of CCR2+ neutrophils and inflammatory monocytes to the skin. In mast cell-deficient Mcpt5Cre+ x Rosa26DTA and in mice in which mast cells do not produce IL-10 (Mcpt5Cre+ x Il10fl/fl) the capacity for NKA to reduce cytokines and chemokines was diminished. Lastly, in skin explant NKA diminished IL-1b and CCL2, genes upregulated after DNFB-treatment. Collectively, these data suggest that NKA reduces inflammation in the skin in a mast cell/IL-10 dependent manner and demonstrate a novel regulatory pathway for NKA in the skin that acts on mast cells to unleash their potential to reduce cutaneous inflammation.
Mice harboring a human Mediterranean G6PD gene variant causing G6PD deficiency have decreased B1b cells and atheroprotective IgM, and increased atherosclerosis.
Authors: Andrews Patrick , Srikakulapu Prasad , Zhou Yilun , Ransegnola Brett , Blackburn Cassidy , Dennis Emily , Loeb Skyler , Ozsvar Kozma Maria , Hay Ariel , Root Katherine , Marshall Melissa , Binder Christoph , Zimring James , McNamara Coleen 
Published: November 2024

Abstract
Epidemiological studies suggest a role of glucose 6-phosphate dehydrogenase deficiency (G6PDd) in exacerbated atherosclerosis. To date, no studies have established a causal relationship between the two. We herein report the first direct evidence of G6PDd causing atherosclerosis and deficits in B cells. Mice harboring the human Mediterranean G6PDd variant (hG6PDMed-) or the nondeficient human G6PD allele (hG6PDND) were rendered hyperlipemic via PCSK9 AAV and 12 weeks of high fat diet (HFD). Sudan-IV en face staining revealed increased atherosclerosis in the G6PDd mice when normalized to non-HDL cholesterol (hG6PDND n=9, hG6PDMed- n=8, P=0.0339). Next, flow cytometry was used to identify alterations in immune cells involved in atherosclerosis and revealed reduced atheroprotective B1b cells (CD19+B220-IgM+CD5-) in the peritoneal cavity and spleen (hG6PDND n=9, hG6PDMed- n=8, P=0.0111 & P=0.0207). B1b cells secrete atheroprotective IgM to oxidation-specific epitopes (IgMOSE), and thus ELISA was used to measure IgMOSE levels. Despite normal total IgM levels compared to non-G6PDd mice, G6PDd mice had reduced IgMOSE to CuOx-LDL and PC-BSA when normalized to total IgM at baseline (hG6PDND n=8, hG6PDMed- n=7, P=0.0289 & P=0.0022) and at 2 weeks of HFD (hG6PDND n=9, hG6PDMed- n=6, P=0.0016 & P=0.0028). Together, these data demonstrate that G6PDd worsens atherosclerosis and suggest that this effect may be partly mediated by associated deficits in B1b cells and IgMOSE.
Genetic screening to identify novel targets enhancing regulatory T-cell (Treg) phenotype.
Authors: Walsh Ariane , Kramer Marina , Wahmes Jan , Komadina Dana , Luu Sabine , Cornils Hauke , Dangl Markus , Cernecka Hana 
Published: November 2024

Abstract
Tregs possess immunosuppressive properties that can restore and maintain immune homeostasis via various suppressive mechanisms. A marker of Tregs is the transcription factor Forkhead box P3 (FoxP3), which regulates Treg development and function. Autoimmune patients have shown dysfunction or reduction in FoxP3+ Tregs, highlighting the critical role of these cells in preventing pathogenic immune responses. There is great interest in developing medical interventions against autoimmune diseases by enhancing Treg function and/or numbers. Currently, a major limitation to generate efficient Treg therapies is the lack of selective targets as starting points for drug discovery.
To identify novel Treg targets, Evotec developed a CRISPR/Cas9-based arrayed screening platform using human CD4+ T-cells. About 8,000 genes were screened with a FACS read-out and the effect of gene knockout (KO) on the frequency of FoxP3+ Tregs was assessed. We identified ~450 targets where the KO led to an increase of FoxP3+ cells within the CD4+ T-cell compartment, indicating their potential for drug development starting points. In addition to FoxP3, the effect of KO on CD4+ conventional as well as Treg viability, proliferation and markers such as, CTLA-4, PD-1 or CD25 was monitored. The hits are being confirmed and the impact of the target KO on Treg function and phenotype assessed in a panel of in vitro assays. The goal is to identify novel Treg targeting product candidates to treat autoimmune diseases.
Regulation of interferon alpha production by MAGUK-family protein CASK in H5N1-infected Macrophage
Authors: Huang Jing-Ying , Sung Pei-Shan , Hsieh Shie-Liang 
Published: November 2024

Abstract
To investigate the involvement of CASK, a MAGUK-family scaffold protein, in H5N1-infected macrophages, we generated mice with myeloid-specific CASK knockout (CASKfloxLysMCre). Using bone marrow-derived GM macrophages (BMDM-GM), we explored the impact of CASK KO on H5N1-induced cytokine storm.
We found that CASK KO in H5N1-infected BMDM-GM selectively reduces interferon alpha production by half, attributed to interference with Ifna2/4 mRNA nuclear export, as confirmed by ELISA and RNA FISH. We also found that H5N1-triggered CASK nucleus translocation mirrors responses to RIG-I and MDA-5 ligand stimulation. LC-MS-MS analysis identified HCK as a promoter of CASK phosphorylation at the S395 site, likely through CDK5. Co-overexpression of EGFP-CASK with S395A mutation and constitutively active HCK resulted in significantly reduced nuclear EGFP signal. CASK Interactome analysis (by IP-LC-MS/MS) and NCBI-interactome meta-analysis indicated that nuclear CASK is recruited to the Ifna promoter by IRF7-binding CCT4. This facilitates co-transcriptional loading of the CASK complex onto Ifna transcripts and supports its CRM-1-dependent nuclear export during H5N1 infection. In vivo studies demonstrated that myeloid-specific CASK KO exacerbates H5N1-induced lung pathology. Our results strongly suggest that CASK plays a crucial role as a host factor, with its nucleus translocation in response to H5N1 infection promoting the nuclear export of anti-viral Ifna mRNA.
The immunological aftermath of SARS-CoV-2: How prior infection impacts subsequent pulmonary injury
Authors: Roberts Lydia , Wehrly Tara , Schwarz Benjamin , Bohrnsen Eric , Shaia Carl , Bosio Catharine 
Published: November 2024

Abstract
Following pulmonary insults there are persistent alterations in the lung that influence responsiveness to subsequent injury. Over 600 million individuals have survived SARS-CoV-2 (SCV2) infection. Thus, the impact of SCV2 on the ability of the newly remodeled lung to handle various pulmonary challenges is of significant interest and therapeutic importance. Using the K18-hACE2 mouse model, we found that SCV2 resulted in an expanded pool of recruited alveolar macrophages along with persistent lymphocytic perivascular cuffs up to 90 days post-infection in the absence of detectable viral antigen. SCV2 resolved lungs had only modest changes following an OVA-specific allergic response compared to mock infected controls. In contrast, despite no differences in bacterial burdens, SCV2 experienced lungs had significantly less inflammation following infection with Bordetella pertussis compared to mock controls. However, this was not consistent for all bacterial driven inflammatory responses. Specifically, we observed an exacerbation of LPS derived inflammation in mice that had recovered from SCV2. The balance of inflammatory responses in these models was associated with specific changes in lipid mediators (LMs). Together, our data reveal the impact of SCV2 infection on the ability of the lung to optimally respond to a variety of inflammation provoking stimuli and underscore the potential role of LMs in the trained innate immune response.
Aryl hydrocarbon receptor ligand supplementation impacts colitis-associated depressive-like behavior
Authors: Bogdon Raymond , Roark Kasie , Saxena Archana , Mitchell Chandani , Hailey Michele , Staley Shanieka , Nagarkatti Prakash , Nagarkatti Mitzi , Busbee Philip 
Published: November 2024

Abstract
Colitis is an inflammatory bowel disease (IBD) with increased incidences of depression in patients. The mechanisms that define this comorbidity remain poorly understood. Our lab previously showed supplementation with indole-3-carbinol (I3C), an aryl hydrocarbon receptor (AhR) ligand, can reduce disease severity in models of colitis. We aimed to determine the behavioral impact I3C supplementation has on colitis-induced depression via alterations in the gut metabolome. Colitis was induced in using the dextran sodium sulfate (DSS) method and treatment groups were given a regimen of 40 mg/kg I3C as previously reported. Untargeted metabolomic studies revealed quinolinic acid (QA), a metabolite produced by the kynurenine (KYN) pathway and linked to depression, was found to be significantly reduced in I3C-treated mice when compared to colitis controls. To determine the effects of I3C and QA modulation on depressive-like behavior, colitis mice were treated with either I3C or an inhibitor of a major enzyme involved in QA production. Depressive-like behavior was measured using the Tail Suspension Test (TST) method, along with evaluation of neural biomarkers of depression (dopamine, BDNF, GFAP) and stabilization of the blood brain barrier (BBB). Results showed I3C reduced depressive-like behavior and altered select biomarkers associated with depression. These studies provide evidence that AhR can be a potential therapeutic target for both colitis and colitis-associated depression.
Determination of IgA antibodies colostrum against Mycobacterium antigens.
Authors: Romero-Ramírez Héctor , Baltierra-Uribe Shantal Lizbeth , Pedraza-Romero Martha Raquel , Barreto-Lesmes Natalia , Sandoval-Montes Claudia , Santos-Argumedo Leopoldo 
Published: November 2024

Abstract
Colostrum, the first milk secretion, is characterized by a high concentration of immunologically active molecules with neutralizing and antibacterial capabilities. Mycobacterium tuberculosis is a microorganism primarily infecting the lungs but can also affect non-pulmonary tissues. Despite a BCG vaccination coverage exceeding 97% in Mexico, pulmonary and meningeal tuberculosis cases have shown a yearly increase. This study aimed to determine the presence of IgA antibodies against soluble extracts from H37Rv M. tuberculosis and BCG antigens in colostrum samples from Mexican mothers. Quantification of IgA subclasses and colostrum reactivity to Mycobacterium were performed via ELISA using 106 colostrum samples from healthy Mexican mothers. Results indicated that 87.7% of the analyzed colostrum samples contained total IgA antibodies, with 77% exhibiting IgA2 antibodies against H37Rv, of which 22.6% and 17.9% showed high titers, respectively. Among the H37Rv antigen-positive samples, 80% recognized the BCG antigen. In conclusion, most of the analyzed colostrum samples from women contained IgA antibodies against M. tuberculosis antigens and demonstrated BCG reactivity. Most of these antibodies belonged to the IgA2 subclass, suggesting a potential protective effect against tuberculous meningitis in nursing infants.
Chimeric Antigen Receptor T-Cell Recruitment and Killing can be Evaluated on an Organ-Chip model system
Authors: MEHTA ANITA , Jacob Irene , Downs Isaac , Kanellias Marianne , Cespedes Adriana , Heng Anthony , Ewart Lorna 
Published: November 2024

Abstract
Objective. Chimeric antigen (CAR) T-cell therapies for solid tumors are challenging to develop due to the lack of human relevant models that adequately capture the mechanisms of CAR T-cell recruitment-a critical yet often overlooked rate-limiting step. The current study aimed to develop a novel system for investigating both the recruitment and killing capacity of CAR T cells in Organ-Chips.
Methods. Chip-S1™ Stretchable Chips (Emulate) were seeded with a human HER2+ non-small cell lung cancer cell line (NSCLC) in the top channel and lung microvascular endothelial cells in the vascular (bottom) channel to create a tumor-vascular interface. Both channels were primed with an NSCLC-specific mix of chemokines and cytokines. HER2-specific CAR T cells were perfused through the vascular channel. Total CAR T-cell recruitment into the top channel and CAR T-mediated killing were measured over 48 hours. The IL-2 cytokine was also co-administered with CAR T cells as immunotherapy.
Results. HER2 CAR T cells demonstrated significant antigen-specific killing when recruited to the top channel as measured by caspase activity and cytokine release compared to the control. Furthermore, IL-2 co-therapy increased CAR T-cell recruitment and antigen-specific killing compared to the control group.
Conclusion. These findings suggest that human-centric Organ-Chips can evaluate CAR T-cell therapies for solid tumors by integrating the critical rate-limiting steps of CAR T recruitment and killing.
Transcriptional profiling of primary human-skin derived mast cell activation by various MRGPRX2 agonists and inhibition with EVO756
Authors: Bagchi Sreya , Patel Jeegar , Harden Jamie 
Published: November 2024

Abstract
Mas-related G-protein coupled receptor X2 (MRGPRX2) is a GPCR expressed predominantly by mast cells and has been implicated to play an important role in diseases such as chronic spontaneous urticaria, atopic dermatitis, and asthma. Cationic ligands, either endogenous or exogenous, can activate the receptor and induce IgE-independent mast cell activation, resulting in allergic responses and non-histaminergic itch. We have previously shown that EVO756, a novel small molecule antagonist of MRGPRX2, demonstrates potent, concentration dependent inhibition of MRGPRX2 mediated activation of transfected CHO-cells, ROSA and LAD2 mast cell lines, and primary human skin-derived mast cells. Furthermore, EVO756 does not affect mast cell viability. Here, we describe, for the first time, the transcriptional landscape of primary human skin-derived mast cells cultured in the presence of several MRGPRX2 agonists and analyze how the mast cell phenotype is altered in response to MRGPRX2 inhibition by EVO756.
Exploring Pre-Pandemic SARS-CoV-2-Specific B Cell Responses in the West African Population.
Authors: Sharma Pragati , Hurtado Jonathan , Messmer Terrence , Briney Bryan 
Published: November 2024

Abstract
The emergence of the novel coronavirus, SARS-CoV-2, during the COVID-19 pandemic prompted investigations into pre-existing immunity in human populations. Despite being a new coronavirus, individuals had previous exposure to common seasonal human coronaviruses. In this study, we examined SARS-CoV-2-specific memory B cells in six pre-pandemic Lassa virus-infected patients from West Africa.
By incorporating a SARS-CoV-2 Spike Antigen (WA-1 strain) into a broader antigen panel that included Lassa virus GPC and a control, we identified and sorted antigen-positive B cells. Further analysis revealed a distinct population of SARS-CoV-2-specific memory B cells. Native paired monoclonal antibody (mAb) sequences were recovered from 598 SARS-CoV-2 Spike-positive B cells, demonstrating a surprisingly high frequency of pre-existing SARS-CoV-2-specific B cells.
Remarkably, Spike-specific mAbs were recovered from all six donors, each contributing to at least one expanded clonal lineage. Notably, most IgM antibodies belonged to the same lineages. These findings suggest a notable prevalence of pre-existing immunity to SARS-CoV-2 in pre-pandemic patients, potentially influencing susceptibility to the virus and contributing to adverse COVID-19 outcomes. This study provides insights for risk management and supports the development of universal coronavirus vaccines by highlighting the complex interplay between pre-existing immunity and susceptibility to SARS-CoV-2.
Whole slide imaging modes for Imaging Mass Cytometry reveal cellular diversity of the tumor immune microenvironment in mouse glioblastoma
Authors: Motta Vinicius , Raza Qanber , Zabinyakov Nick , Pfister Thomas , Parsotam Nikesh , Howell David , Lim Liang , Loh Christina 
Published: November 2024

Abstract
Mouse models are used for translational research and permit detection of therapeutic-induced immune-related modification in the tumor immune microenvironment (TIME). Imaging Mass Cytometry™ (IMC™) is a spatial biology technique capable of quantitative evaluation of multiparametric tumor and immune cell composition in brain tissue.
Here we demonstrate the application of a 40-marker panel composed of tumor and immune markers on mouse normal and glioblastoma (GBM) tissue using two new whole slide imaging methods for IMC. Tissue mode (TM) rapidly scans the entire tissue followed by pixel-based analysis (PBA) to uncover spatial distribution of relevant markers. Preview mode (PM) samples the entire tissue at predefined spacing to rapidly capture a low-resolution image of all expressed markers, improving informed identification of areas for high-resolution cell mode (CM) imaging on the same slide.
TM imaging with PBA provided quantitative spatial expression patterns of tumor and immune markers across the GBM tissue. Necrotic cores, areas with high immune infiltration, extracellular matrix deposits, and activated tumor cells were quantified. PM in combination with CM imaging and single-cell analysis revealed extensive tissue vascularization, replicating tumor cells, and a variety of infiltrating immune cells.
Overall, we highlight the capability of IMC to provide quantitative insights into the spatial biology of the TIME.
For Research Use Only. Not for use in diagnostic procedures
RNA sequencing reveals differential gene expression by hypoxia in γδ T cells.
Authors: Park WonHyung , Lee Heung Kyu 
Published: November 2024

Abstract
Hypoxia not only enhances tumor survival but also inhibits the anti-tumor response. Particularly, hypoxia is known to promote exhaustion of T cells. So, overcome hypoxia becomes imperative, especially in T cell therapy. One prominent T cell therapy is CAR T cell therapy. However, it has limitation because of autologous applications. Recent attention has shifted towards γδ T cell therapy, especially when autologous options are limited by senescence or expansion suppression in tumor patients. In the context of γδ T cell therapy, investigating the effects of hypoxia on γδ T cells is pivotal for optimizing their functionality. Therefore, this study aims to investigate how γδ T cells change under hypoxic conditions. The ultimate goal is to uncover specific targets that can mitigate the negative consequences of hypoxia, thereby improving the overall efficacy of γδ T cell-based therapies.
γδ T cells is stimulated with γδ TCR antibodies for ex vivo expansion. Subsequently, total RNA is purified from γδ T cells following a 24-hour culture under hypoxic condition (1.0% O2). Bulk RNA sequencing is then conducted on the collected samples.
Through bulk RNA sequencing, it is established that hypoxia inhibits the proliferation of γδ T cells. This finding holds particular significance in the context of therapy maintenance. Furthermore, the analysis reveals upregulation in cytokine genes, notably Il10 and Tgfb3, suggesting a potential regulatory role in anti-tumor immune responses.
IL-33 mediated regulation of Tregs is tissue specific and dispensable in the omental visceral adipose tissue.
Authors: Desai Sweta , Meza-Perez Selene , Liu Mingyong , Randall Troy 
Published: November 2024

Abstract
The omentum, a visceral adipose tissue in the peritoneal cavity, is a frequent site of metastasis for gastric, ovarian and various other cancers. Currently available therapies are ineffective in providing long-term tumor control, with a higher plausibility for relapse. The rapid growth of tumors in the omentum is supported by the expansion and accumulation of immunosuppressive Tregs. Additionally, we detect persistent and continuously expanding tumor specific CD8 T cells despite Treg accumulation in the omentum. Omental Tregs are a specialized population of visceral adipose tissue associated Tregs (VAT-Tregs). Some of them express the interleukin-33 receptor ST2, suggesting IL-33 produced in the omentum promotes Treg mediated immunosuppression. However, our findings show that absence of IL-33 mediated signaling in VAT-Tregs does not alter their accumulation or expansion in the tumor bearing omentum. Moreover, we demonstrate that absence of IL-33 signaling in the omental VAT-Tregs does not reduce tumor growth, enhance the persistence of tumor specific CD8 T cells, or CD8 T cell mediated tumor killing. Our results show that IL-33 is dispensable for Treg maintenance or accumulation in the omental visceral adipose tissue. We are investigating the pathways involved in regulation of Tregs in the omentum.
Gelatinases are important for CD4+ T-cell metabolic reprogramming
Authors: Protz Ella , Onwuha-Ekpete Lillian , Fields Gregg 
Published: November 2024

Abstract
Matrix metalloproteinases (MMP) are a family of proteolytic enzymes essential for various processes such as development and healing. Alternatively, the dysregulation of the gelatinases, MMP-2 and MMP-9, is implicated in immune cell migration and inflammation associated with multiple sclerosis (MS). MS is an autoimmune disease in which CD4+ T-cells play a key role both in humans and its animal model, experimental autoimmune encephalomyelitis (EAE). CD4+ T-cells are known to rely on both oxidative phosphorylation and aerobic glycolysis during various stages of activation. We have recently demonstrated that gelatinase inhibition can impact CD4+ T-cell activation and mitigate EAE progression. The present study utilized the Seahorse assay to characterize the cellular energetics of CD4+ T-cells in response to gelatinase inhibition. The resultant data suggests that gelatinases are important for both oxidative phosphorylation and the effective induction of glycolysis in CD4+ T-cells.