bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enrichement of type I interferon signalling in the colonic ILC2 under inflammatory condition
Authors: Mikami Yohei , Emi Irie , Rino Ishihara , Ichiro Mizushima , Kanai Takanori 
Published: August 2023

Abstract

Background
Group2 innate lymphoid cells (ILC2s) are known to contribute to a frontline defence against parasites and formation of allergic diseases, but little is known about the roles of ILC2s under intestinal inflammation including intestinal bowel diseases (IBD). In this study, we investigate the characteristics of ILC2s in health and colitis.

Methods
ILC2s were analysed in the mice with or without treatment of dextran sulphate sodium (DSS)-induced colitis, where surface markers and cytokine production was assessed by flow cytometry. The global gene expression of ILC2 in DSS-induced colitis and control mice were analysed by bulk RNA-sequencing (RNA-seq). Lastly, the mice was treated with anti-IFN-alpha/beta receptor1 (IFNAR1) antibody and then challenged to DSS-induced colitis.

Results
The number of ILC2s in the gastrointestinal tract was larger than that of spleen and liver in the healthy mice. At day 7 of DSS-induced colitis, IL-5 production from ILC2s was significantly reduced in flow cytometry data, and this was accompanied by enrichment of type I interferon (T1IFN) signalling in colitic ILC2 in RNA-seq data. Neutralisation of T1IFN signalling by anti-IFNAR antibody significantly increased severity of colitis.

Conclusion
Our data suggest that downregulation of Th2 cytokines and enrichment of T1IFN signatures in colitic ILC2s. T1IFN may stimulate ILC2 to induce colitis suppressive functions.

This study was funded by the Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Transformative Research Areas(B): 21H05123; Advanced Research and Development Programs for Medical Innovation (AMED-CREST: 21gm1510002h0001, and 20gm1210001h0001; the Practical Research Project for Rare/Intractable Disease: 21ek0109556h0001)
Intestinal epithelial expression of neutral ceramidase determines severity of chemical colitis in mice
Authors: Deng Zhongbin , Xu Zhishan , Lei Chao , Sun Rui , Sriwastva Mukesh , Tan Yi 
Published: August 2023

Abstract
Sphingolipids are important eukaryotic signaling molecules and regulate inflammation and immunity. Elevated levels of sphingolipid metabolites, e.g. S1p, are found in many chronic inflammatory disorders, including inflammatory bowel disease (IBD). Neutral ceramidase (NcDase) is a key regulator of ceramide, the central intermediate in the sphingolipid metabolism. The contribution of intestinal epithelial cells (IEC) NcDase to IBD is not well understood. Here we demonstrate that mice with IEC-specific loss of NcDase (NcDaseΔIEC) developed less severe DSS-induced colitis than control mice. Deletion of IEC NcDase leads to increase of antimicrobial peptides, anti-inflammatory cytokines and regulatory T (Treg) cells; and reduction of dysbiosis and gut inflammation. Mechanistically, IEC NcDase is required for the generation of ganglioside GD3, which controls the enteric immune response by activating Treg cells and preventing activation of T-helper (Th)17 cells. Administration of dietary GD3 reduces mucosal inflammation. Thus, GD3 is a key epithelial-derived glycosphingolipid that regulates distinct subsets of intestinal CD4+ T cells during inflammatory conditions, a finding with potential implications for treatment of chronic inflammatory disorders.
Supported by grants from NIH R21AI159194, R01 DK115406, and R01DK131442 (Z.D.).
Investigation of the aryl hydrocarbon receptor’s role in colitis as it relates to observed disease-specific sex differences
Authors: Mitchell Chandani , Saxena Archana , Staley Shanieka Caroline , Hailey Michelle , Bogdon Raymond K , Roark Kasie , Nagarkatti Mitzi , Nagarkatti Prakash S , Busbee Phillip B 
Published: August 2023

Abstract
Colitis is an inflammatory bowel disease (IBD) characterized by chronic inflammation in the colon. Studies suggest that there are differences between males and females regarding colitis disease progression, severity, and response to treatment. However, there lacks clear understanding, and some inconsistencies, on what these differences are attributed to. Previously, our reports showed that activation of the aryl hydrocarbon receptor (AhR) ligand was able to reduce colitis-induced disease severity, mainly through innate lymphoid type 3 cells (ILC3), in an interleukin-22 (IL-22) dependent manner. However, noted sex differences in disease severity and treatment response were observed in the animal models of colitis. In the current study, we generated cell-specific conditional knockout (KO) mice that had AhR deficiency in Rorc-expressing immune cells (ILC3s). We induced colitis using the dextran sodium sulfate (DSS) model in wildtype (WT) control mice and conditional mouse strains with AhR deficiency in Rorc-expressing immune cells (AR mice). Weight data and serum inflammatory biomarker assays showed that male mice developed a more severe colitis compared to females, with the male AR mice even worse. In addition, while control WT DSS female mice had less severe colitis than male counterparts, AR DSS female mice had just as severe disease as males, suggesting AhR, in part, was necessary to help promote less disease severity in females. Collectively, these results show that there are sex differences in colitis induction between male and female mice, and also that these differences might in part be due to some intrinsic sex-linked expression of AhR in different cell types.
The studies were supported in part by NIH grants P20GM103641.
IL10 producing regulatory T cells support fecal microbiota transplant (FMT)-mediated clearance of Clostridioides difficile Infection
Authors: Alam Md Zahidul , Maslanka Jeff R , Denny Joshua E , Bittinger Kyle , Abt Michael C 
Published: August 2023

Abstract
Fecal microbiota transplantation (FMT) is an effective therapy for treating recurrent C. difficile infection. However, the FMT mechanism of action is poorly understood, and the implementation of FMT therapy is limited. A previous study from our lab identified a critical role of CD4+ FoxP3+ regulatory T cells (Treg) in enabling FMT engraftment and subsequent resolution of C. difficile infection. Here, we investigated the cellular and cytokine network regulated by Treg cells to support FMT-mediated resolution of C. difficile infection. On the day of FMT, C. difficile infected mice have increased IL10+ Treg cells in the colon compared to uninfected mice. Further, mice depleted of IL10 and its signaling pathway are defective in resolving C. difficile following FMT, mirroring our observation in Treg-depleted FoxP3-DTR mice. The depletion of Treg cells in C. difficile-infected FoxP3-DTR mice induces a robust type 1 immune response in the colon characterized by IFNγ producing Th1 and type 1 innate lymphoid cells. This immune response drives an increased infiltration of iNOS+ neutrophils into the large intestine. Furthermore, neutralization of IFNγ following Tregs depletion decreases iNOS+ neutrophils in the large intestine, enabling the FMT to resolve C. difficile infection. Last, the depletion of neutrophils in Treg-depleted mice restores the efficacy of FMT to resolve C. difficile. Combined, we provide a mechanism by which IL10+ Treg cells serve as a negative regulator of an IFNγ-neutrophil intestinal inflammatory circuit, thereby supporting the colonic environment to be receptive to FMT and resolution of C. difficile. These results provide a mechanistic example of how immunity can dictate the efficacy of microbiome-based therapies.
Supported by grants from NIH (R01 AI158830, R21 AI164385)
Prostaglandin E2 signaling via its receptor EP4 on macrophages protects against acute colitis by preserving intestinal barrier function
Authors: Murray Page , Kelsall Brian 
Published: August 2023

Abstract
The inflammatory bowel diseases are increasing in incidence and cause significant morbidity; however, disease pathogenesis is poorly understood. Prostaglandin E2 (PGE2) may protect against intestinal inflammation and its receptor EP4 is highly expressed on colon macrophages, cells essential for immune homeostasis, suggesting PGE2 signaling on macrophages may prevent gut inflammation.
To address this, we induced DSS colitis in CX3CR1CRE/+EP4fl/fl (Cre+) mice, which lack EP4 on macrophages, and in their Cre- wildtype littermates. Cre+ mice developed worse colitis than the Cre- based on clinical scores, including increased rectal bleeding that implies major intestinal barrier disruption, a key mechanism of colitis. Cre+ mice have higher intestinal permeability early during inflammation, further supporting intestinal barrier disruption as a mechanism of increased disease severity in Cre+ mice.
To address the protective mechanisms of PGE2, we performed scmRNA sequencing on colon cells of the Cre+ and Cre- mice following DSS treatment. We identified monocyte and macrophage populations, with macrophages (MPs) clustering into CD11c+ MPs expressing inflammatory cytokines, and CD169+ MPs associated with inflammation resolution and wound-healing. During colitis there was a progressive increase in the proportions of inflammatory monocytes and a loss of CD169+ MPs that was accelerated in Cre+ mice. Further, the proportion of CD11c+ MPs was increased in Cre+ mice. These data are the first detailed scmRNA analysis of immune cell populations in DSS colitis and suggest that PGE2 signaling through EP4 may protect non-inflammatory macrophage populations during inflammation, contributing to maintenance of the intestinal barrier.
High salt diet impairs the protective immunity against mucosal HSV-2 infection
Authors: Kwon Myoung Seung , Park Jang Hyun , Lee Heung Kyu 
Published: August 2023

Abstract
Herpes simplex virus type2 (HSV-2) is the main cause of genital ulcer disease. Because of asymptomatic viral shedding, HSV-2 is highly prevalent and infected nearly 500 million people worldwide. Recently, excessive sodium intake has threatened global health. It is well known that excessive sodium intake affects various diseases by altering gut microbiome. However, the effect of excessive sodium intake on vaginal microbiome and vaginal immune system is unknown. In this study, we investigate the effects of high salt diet on vaginal microbiome and HSV-2 disease progression. We found that high salt diet compromises the anti-viral immunity in a mouse HSV-2 model. In 16S ribosomal RNA sequencing of vaginal microbiome, high salt diet results vaginal dysbiosis. High salt diet induced vaginal dysbiosis promotes neutrophil infiltration in vaginal tissue and vaginal lumen. In addition, Single-cell RNA analysis of vaginal neutrophil reveals dominant increase in neutrophil expressing MMPs that could damage vaginal tissue. Our findings suggest that high salt diet related vaginal dysbiosis impairs antiviral protection via neutrophil recruitment.
Death of Tonsillar B Cells by NETosis
Authors: Bukhari Ameera M , Knight Katherine L 
Published: August 2023

Abstract
Regulating B cell death is essential for generating antibodies and maintaining immune tolerance. B cells can die by apoptosis, and we report that human tonsil B cells, but not peripheral blood B cells also die by NETosis. This cell death is density-dependent, characterized by the loss of cell and nuclear membrane integrity, release of reactive oxygen species, and chromatin decondensation. Tonsil B cells secrete high levels of TNF, and inhibiting TNF prevented chromatin decondensation. By in situ fluorescence microscopy, B cell NETosis, as identified by the hyper citrullination of Histone 3, was localized to the light zone (LZ) of germinal centers in normal tonsil and overlapped with the B cell marker CD19/IgM. We propose a model in which stimulation of B cells in the LZ induces NETosis, driven in part by TNF. We also provide evidence that NETosis of tonsil B cells may be inhibited by an unidentified factor in tonsil. The results describe a previously unidentified form of B cell death and suggest a new mechanism to maintain B cell homeostasis during immune responses.
Role of MR1-driven signals and amphiregulin on the recruitment and repair function of skin MAIT cells
Authors: de Poulpiquet du Halgouet Anastasia , Darbois Aurélie , Alkobtawi Mansour , Mestdagh Martin , Premel Virginie , Colombeau Ludovic , Rodriguez Raphaël , Zaiss Dietmar , Aractingi Selim , Golub Rachel , Lantz Olivier J , Salou Marion 
Published: August 2023

Abstract
Epithelial repair is crucial to reestablish adequate barrier integrity and function after tissue injury. In both mice and humans, MAIT (Mucosal associated invariant T) cells involvement in tissue repair is substantiated by their upregulation of tissue repair mediators following TCR-mediated activation. Additionally, in the absence of ɣδT cells, MAIT cells were shown to promote wound healing. By using both our congenic B6-MAITCAST strain (10× more MAIT cells than B6 mice) and a skin excision mouse model preventing epithelial contraction (mimicking the human repair process), we sought to uncover the underlying mechanisms of MAIT cell tissue repair function.
Firstly, our scRNAseq analysis revealed that steady-state skin MAIT cells readily express a repair program. Additionally, we show that MAIT cells have a pro-repair function as their presence promotes keratinocyte proliferation, thereby accelerating healing. Using parabiosis, skin graft and the Kaede models, we found that, contrary to the resident ɣδT cells, MAIT cells are rapidly recruited into the inflamed skin. Parabiosis of MR1+ and MR1− mice and MR1− skin grafted onto MR1+ mice, showed that this recruitment was MR1-independent (MR1: antigen presenting molecule of MAIT cells). Instead, this recruitment relied on the chemokine receptor CXCR6 and its ligand CXCL16. Lastly, using conditional knock-out mice we demonstrated that the secretion by MAIT cells of the amphiregulin growth factor promotes skin wound healing.
Overall, our study provides mechanistic insights into MAIT cell biology in the context of injury. Comprehension of these processes provides clues into the biology of related innate subsets and highlights potential new therapeutic avenues for wound healing.
Institut National de la Santé et de la Recherche Médicale (OL)
Institut Curie (OL)
Agence Nationale de la Recherche Grant MAIT ANR-16-CE15-0020-01 (OL)
Agence Nationale de la Recherche Grant MAITrepair ANR-20-CE15-0028-01 (OL)
Agence Nationale de la Recherche ANR-10-IDEX-0001-02 PSL (OL)
European Research Council (ERC-2019-AdG-885435) (OL)
Fondation pour la Recherche Médicale, grant number FDT202106013036 (AdH)
Peripheral viral detection induces an antimicrobial program in olfactory bulb immature neurons
Authors: Garcia Benjamin J , Kraus Aurora , Salinas Irene 
Published: August 2023

Abstract
The immune and nervous systems have co-evolved to sense and eliminate danger. Neuroimmune interactions occur bidirectionally between the periphery and the central nervous system (CNS) in physiological and disease states. In the olfactory periphery, olfactory sensory neurons (OSNs) are exposed to environmental pathogens and project their axons onto the olfactory bulb. Previous studies have shown that the olfactory bulb participates in the immune response when a pathogen directly invades the CNS. We hypothesize that immune responses occur in the olfactory bulb in response to peripheral pathogen detection by OSNs. We have leveraged zebrafish as a model organism to study neuronal signaling in response to peripheral viral stimuli without infection of the olfactory bulb. Using single cell RNA sequencing we demonstrate that microglia respond to peripheral viral stimuli within 15 min, while a population of immature neurons expressing the neurotransmitter adcyap1 (PACAP-38) and scg5 (secretogranin V) expands within one day. PACAP-38 is a highly conserved neuropeptide with immunomodulatory and antimicrobial functions in mammals, suggesting that immature neurons participate in the antiviral state of the olfactory bulb. Neuronal transcriptional responses to olfactory viral stimuli were coupled to neuronal activation in the olfactory bulb. Furthermore, neuronal changes are translated into behavioral responses, as shown by a reduced mean velocity in an open field test after one day in virally treated compared to control animals. Taken together, these results indicating that peripheral viral stimuli induce a neuropeptide-based antimicrobial program in the olfactory bulb of adult zebrafish.
Premature Death of Sleep Fragmented Males is Associated With An Accumulation of Reactive Oxygen Species In The Gut
Authors: Moriarty Alina K , Keeter W Coles , Kirk Cassandra Lee , Stahr Natalie Ann , Sanford Larry , Galkina Elena 
Published: August 2023

Abstract
70 million Americans report insufficient sleep due to work or lifestyle choices. Decreased duration and poor sleep quality are associated with high rates of CVD and is strongly associated with increased risk of all-cause mortality. The effects of gender have been studied in CVD, but the impact of sexual dimorphism in Sleep Fragmented (SF)-associated atherogenesis is not known. Recently, we found that high fat diet (HFD) fed male Apoe−/− mice on SF experienced premature death. 80% of males did not survive past week 2 of SF, while 80% of females survived past week 8 under same conditions. We sought to identify a cause of premature death in HFD fed male Apoe−/− mice following sleep fragmentation. Interestingly, HFD fed male Ldlr−/− mouse did not experience premature death when SF, suggesting that ApoE absence, not hyperlipidemia, plays an essential function in the induction of death. Furthermore, oral 17β-Estradiol was able to rescue HFD fed male Apoe−/− mice from SF induced premature death. Compared to controls, SF, HFD fed male Apoe−/− mice had decreased body weight and increased circulating LPS, ROS and cell-free dsDNA at death. Finally, the small intestines SF, HFD fed male Apoe−/− mice had increased ROS and decreased tight junction protein (Tjp1) expression, suggesting compromised intestinal barrier function. Our data suggests that a combination of known risk factors for CVD (male sex, ApoE genotype, and HFD) results in premature death when combined with inadequate sleep. Furthermore, our data suggests that this specific combination of co-factors might result in premature death due to a systemic increase in oxidative stress, possibly caused by leakage of LPS from the gut, which may be alleviated through the administration of 17β-Estradiol.
Supported by project–initiation funds from the Center for Integrative Neuroscience and Inflammatory Diseases and AHA Predoctoral (20PRE35180156)
Using a CRISPR based screen to identify genes dysregulated in AML by T-cells
Authors: Pandita Ravina , Kosaka Yoko , Burns Faith , Lind Evan F 
Published: August 2023

Abstract
Acute Myeloid Leukemia (AML) is the most common hematological malignancy in adults and has a 35% survival rate at 5 years. Genetic heterogeneity makes it a highly lethal disease and patients develop resistance to therapy. Our lab has previously shown that in AML, the percentage of T-cells in the bone marrow at the time of diagnosis has a correlation with overall survival, and we have shown there is a dysregulation of T-cells in AML, suggesting their involvement in immune evasion. However, the mechanism by which T-cell activity contributes to leukemia is not well described. Currently, CRISPR screens are widely used to identify sensitivity and resistance genes involved in T-cell killing of cancer cells. Based on this strategy, we used three AML human cell lines (MOLM 14, OCI-AML2 and HL-60) at several effector to target ratios with T-cells and different treatment conditions and found that a majority of AML cells were killed at the higher T-cell ratio. Additionally, stimulating the T-cells using a bispecific antibody that conjoins T-cells with their targets improved target cell killing, particularly the MOLM 14 cell line. All these cell lines achieved differential killing by T-cells, suggesting depending on mutational background, AML immune evasion gene networks challenge T-cells by different mechanisms. Therefore, our future goal is to use a CRISPR/CAS9 lentiviral library genome-wide screen on the tumor cells followed by T-cell killing to identify genes involved in immune evasion in AML. The results of these experiments will help us to better understand T-cell dysregulation mechanisms in AML, thus providing a roadmap for immune based therapies combined with immune checkpoint blockade.
Supported by grant- NIH 1R01CA262145-01A1
Single-cell landscape analysis of tumor infiltrated immune cell population in FcγRIIB deficiency against glioblastoma
Authors: Ku Keun Bon , Kim Chae Won , Lee Heung Kyu 
Published: August 2023

Abstract
Glioblastoma (GBM) is the most common and deadliest cancer known in the central nervous system. Immune checkpoint inhibitors (ICIs) are effective in clinical trials for hematological tumors. However, the application of ICIs is for GBM patients has not proven to be completely successful. The limitations of the treatments can be attributed to the lack of an understanding of the GBM microenvironment (TME), which consists of heterogeneous tumor stem cells and various types of immune cells. Thus, investigating these tumor-infiltrating immune cells and their functions can provide a better insight to the TME of GBM. In order to accomplish this, we performed single-cell RNA sequencing analysis using the GBM mouse model. From the results, the TME up-regulated the inhibitory FcγR gene, Fcgr2b, on infiltrating immune cells, and this inhibitory receptor suppressed the antitumor immune response against GBM. FcγRIIB is known to mediate immune homeostasis through the binding of the immune complex, and blocking the receptor induces a proinflammatory response. We were able to use the FcγRIIB knockout mouse (KO) and analyze the landscape of tumor-infiltrating immune cells. Tumor-associated microglia portrayed anti-tumoral characteristics in KO mice and we were able to see an upregulation of adaptive immune cell recruiting chemokines. In addition, CD8 T cells showed an enrichment of precursor genes related to T cell exhaustion. With tumor progression, the total immune cell number had increased in KO mouse and CD8 T cells enhanced cytotoxic related genes. We can interpret from these results that FcγRIIB plays a critical role in GBM infiltrating immune cells, and targeting of this receptor could be a possible treatment option for treatment of GBM.
Fast and efficient isolation of human EGFR-positive cells using EasySep&[trade]
Authors: Poon Grace F T , Liang Alice , Chehal Manreet , Eaves Allen C , Louis Sharon A , Antignano Frann 
Published: August 2023

Abstract
Epidermal growth factor receptor (EGFR) is a cell surface receptor normally expressed on most epithelial cells and some hematopoietic cells. Mutation and overexpression of EGFR is a key driver of many human cancers, including lung, breast, as well as colorectal cancers. Circulating tumour cells (CTCs) can also express EGFR, where enrichment of EGFR+ CTCs can provide promising liquid biopsy and prognostic values. EGFR-positive cells can be difficult to isolate because they are present in many tissue types at a wide range of frequencies. We have developed a simple method to isolate human EGFR-positive cells to address this challenge. A549 human lung adenocarcinoma cells expressing EGFR were spiked into human PBMCs at defined frequencies as a model system. Starting with a single-cell suspension with 10% EGFR+ cells, the EGFR-expressing cells were labeled with an antibody complex that linked the EGFR-expressing cells to magnetic particles, then separated using an EasySep™ magnet. Using this method, EGFR+ cells were enriched from 9.9 ± 3.3% to 95.0 ± 2.8% (mean ± SD; n = 14). Protocols have been optimized for different sample sizes and different EGFR+ cell starting frequencies. EasySep™-isolated EGFR+ lung adenocarcinoma cells were viable and proliferative, with fold expansion and Ki67 expression comparable to untreated cells. EasySep™ Human EGFR Positive Selection Kit enables simple and easy isolation of EGFR-positive cells in 15 minutes, facilitating EGFR and cancer research.