bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lymphotoxin β receptor signaling mediates the formation of high endothelial venule-like vessels in atopic dermatitis-like skin lesions in mice
Authors: Kanameishi Shuto , Ono Sachiko , Keith Yuki Honda , Asahina Ryota , Honda Tetsuya , Kabashima Kenji 
Published: January 2023

Abstract
High endothelial venules (HEVs) are specialized blood vessels contribute to the trafficking of lymphocytes into secondary lymphoid organs by expressing a group of L-selectin ligands called peripheral node addressins (PNAd). In lymph nodes (LNs), lymphotoxin β receptor (LTβR) signaling from dendritic cells (DCs) is essential for the formation of HEVs. On the other hand, PNAd+ HEV-like vessels are observed in the skin under certain inflammatory conditions, such as atopic dermatitis (AD). However, the characteristics and the formation mechanism of HEV-like vessels in the inflammatory skin remains unclear. To address these issues, we first identified PNAd+ HEV-like vessels in the skin using a calcipotriol-induced mouse AD model. HEV-like vessels were located in the postcapillary venules abundant in cellular organelles, which were similar to HEVs in LNs. Blockade of LTβR-signaling by administration of LTβR-Fc fusion protein markedly decreased the formation of HEV-like vessels in the AD-like skin. Analysis using published single-cell RNA-seq data of human AD skin revealed that T cells and DCs highly expressed LTB, a gene encoding LTβ. The expression of LTB was highlighted in T cells by RNAscope in both human AD lesions and mouse AD-like skin. The formation of HEV-like vessels in AD-like skin was normal in Ltbfl/fl CD11cCre mice, but was partially impaired in Ltbfl/fl CD4Cre mice. Collectively, HEV-like vessels in the inflammatory skin show similar characteristics to HEVs in LNs, and LTβR signaling, partly by LTβ from T cells, is responsible for their formation.
Immunomodulatory effects of human myeloid-derived suppressor cells expanded from cord blood on dermatophagoides farinae-induced atopic dermatitis in NC/Nga mice
Authors: Kim Chang-hyun , Sohn Hyun-Jung , Hong Seung-Min , Kim Sueon , Jung Soo-Hyun , Lim Bang-Geul , Kim Eun-ah , Jeong Kwang-Hun , Kim Jongil , Park Jaehee , Kim Stan , Kim Tai-Gyu 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) plays an important role in controlling the immune response against cancer and in suppressing autoimmunity and allergic inflammation. However, the therapeutic effect of MDSCs on experimental mouse model of atopic dermatitis (AD) has not been reported. We investigated the therapeutic efficacy of human myeloid-derived suppressor cells expanded from cord blood (hUCB-MDSCs) in dermatophagoides farinae (Df)-induced AD-like skin lesions in NC/Nga mice and explored the mechanisms involved. The administration of hUCB-MDSCs (1×105 and 1×106 cells per mouse) significantly reduced in clinical severity scores and was associated with the reduction of histopathological changes including inflammatory cellular infiltration, epidermal hyperplasis, and hyperkeratosis. hUCB-MDSCs decreased the serum levels of IgE, Th2-mediated cytokines and chemokines. Particularly, the expression of filaggrin and involucrin in the skin lesions was recovered, whereas the expression of keratin-10 and keratin-14 decreased. These immunomodulatory effects of hUCB-MDSCs in a murine model of AD may provide helpful evidence for the clinical development of cell therapies to treat AD.
Macrophage Clearance of Neutrophil Extracellular Traps (NETs)
Authors: Gronevelt J Paige , Pisano Michael , Rumble Julie 
Published: January 2023

Abstract
Neutrophil Extracellular Traps (NETs) are formed from dying neutrophils expelling their contents to capture pathogens and limit the spread of infection. While they are beneficial during infection, overproduction of NETs or the inability to degrade them can be pathogenic. Different inflammatory disorders and autoimmune diseases have been shown to be associated with an increase in NET production. Therefore, it is important to understand the method in which NETs are removed or broken down.
Cayman Chemical has developed an imaging-based assay to sensitively assay NET uptake by phagocytic cells. Imaging was found to be an ideal method to monitor NET clearance in vitro, in part due to the ability to monitor the uptake of NETs kinetically. Both primary monocyte-derived macrophages and differentiated THP-1 cells, which are frequently used as phagocytic cell models, were shown to take up NETs. Putative modulators of NET uptake were tested in the assay.
We aimed to develop a method in which the process of NET uptake can be monitored and quantified. Modulators of NET uptake can be tested in this assay, allowing for further studies toward treating autoimmune disorders that are driven by aberrant NETs.
The splicing factor SRSF6 controls Mycobacterium tuberculosis replication through its role in maintaining mitochondrial homeostasis
Authors: Wagner Allison , Weindel Chi , West Kelsi , Watson Robert , Patrick Kristin 
Published: January 2023

Abstract
Macrophages are the primary host cell that supports the survival and replication of the intracellular bacterial pathogen Mycobacterium tuberculosis (Mtb). We are interested in how macrophage proteins are functionalized during Mtb infection. Global proteomics analysis of Mtb-infected primary macrophages found that many RNA binding proteins of the Serine/Arginine rich splicing family (SRSF) can be differentially phosphorylated, suggesting that these factors regulate splicing decisions during Mtb infection. Knockdown of one such SRSF protein, called SR6, results in basal upregulation of interferon stimulated genes (ISGs) in macrophages. We hypothesized that this phenotype was driven by mitochondrial damage that leaks mitochondria DNA into the cytosol and activates cGAS. Consistent with this idea, we measured increased mitochondrial stress and increased cell death in SR6 KD macrophages. Transcriptomics analysis of these cells revealed upregulation of an alternatively spliced (AS) isoform of the apoptotic factor BAX, which contributes to mitochondrial dysfunction, mtDNA release, and basal ISG expression. While BAX isoforms are known to regulate cell death, this is the first report of BAX AS regulating the type I IFN response. Lastly, we found that SR6 KD macrophages are permissive to Mtb replication, suggesting that controlling cell death at the level of BAX isoform abundance is an important but unappreciated component of the macrophage antibacterial repertoire. Together, our data support a model whereby SR6-dependent AS of Bax triggers mitochondria stress causing mtDNA leakage and type I IFN expression, metabolic dysfunction that primes cells to undergo cell death, and increased susceptibility to Mtb infection.
Defects in macrophage galactose-type lectin (MGL) signaling and pathogenesis of Mycobacterium tuberculosis and Human Immunodeficiency virus co-infection
Authors: Browning Sarah , Naqvi Kubra F , Fan Xiuzhen , Bharaj Preeti , Lisinicchia Joshua , Paez Reina N , Chauhan Sadhana , Huante Matthew , Martinez-Martinez Yazmin Berenice , Endsley Mark , Gelman Benjamin B , Endsley Janice 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb) and Human Immunodeficiency virus (HIV) are important causes of death in infected patients worldwide, and act synergistically to worsen disease in those with co-infection. C-type lectin receptors (CLRs) are an important component of innate immunity that can recognize and respond to pathogen-associated signals. The macrophage galactose-type lectin (MGL, CLEC10) has been described as an immunomodulatory CLR associated with M2 macrophages and regulation of inflammation following infection. Our lab previously identified an important antibacterial role of murine MGL-1 in host immunity to Mtb. More recently we observed that human peripheral blood mononuclear cell-derived macrophages, and THP-1 cells, upregulated MGL following in vitro exposure to Mtb, Mycobacterium bovis BCG, and TLR2 agonists. Silencing of MGL in primary human macrophages permitted greater intracellular Mtb replication. Interestingly, we observed that MGL is suppressed by HIV in differential transcriptome analysis of lung from HIV-infected humanized mice, in lung and spleen of autopsy specimens from HIV-infected decedents, and upon in vitro infection of human macrophages. Using a recombinant human MGL/Fc chimeric molecule we observed that Mtb, and not HIV, directly bound MGL. MGL surface expression was reduced in lung of mice with chronic HIV infection compared to those with Mtb, and variable in co-infected animals. These results indicate that human MGL is an important CLR for pathogen recognition and initiation of protective immunity by host macrophages to Mtb. The suppression of MGL due to HIV may increase susceptibility to Mtb in people with HIV or exacerbate disease progression in those with co-infection.
Supported by grants from NIH (R33 AI138328, R61 AI138328, U24 MH100930)
STING activation under pre-existing inflammatory conditions causes severe skin disease
Authors: Pyclik Marcelina , Bagavant Harini , Papinska Joanna , Araszkiewicz Antonina , Deshmukh Umesh 
Published: January 2023

Abstract
The activation of the stimulator of interferon genes 1 (Sting1) pathway causes rapid induction of type I IFNs. Recently, different STING agonists are being proposed as therapeutics for controlling viral infections like those caused by the SARS-CoV-2. Considering that excessive STING activation leads to autoimmunity, this study was undertaken to investigate the effects of STING activation under pre-existing systemic inflammatory conditions induced by a viral infection. Female BALB/c mice were treated (Tx) with TLR3 agonist poly(IC), which was followed 24h later by subcutaneous injection of STING agonist diamidobenzimidazole (diAbZi). Only diAbZi or vehicle Tx mice were used as controls. Mice Tx with poly(IC) and diAbZi showed the highest systemic levels of type I IFNs, IL-6, and TNFα. Within 10 days, the salivary glands showed increased ILC1, monocytes, and CD8+ T cells. However, glandular function was not affected. Five days after Tx, except for the vehicle controls, all mice showed differing extents of redness, hair loss, and ulceration at the injection site. By day 10, diABZi alone Tx mice showed either normal skin or a small scar formation. In contrast, mice Tx with poly IC and diABZi, showed extensive hair loss, erythema, and ulceration. In addition, hematoxylin and eosin-stained skin sections showed extensive neutrophilic infiltration at the site of ulceration, epidermal thickening, and patchy lymphocytic infiltrate that extended into the subdermal layers. Our data demonstrate that STING activation under systemic inflammatory conditions, such as a viral infection, can lead to severe skin disease. Therefore, caution needs to be exercised in employing STING agonists as therapeutics for treating SARS-CoV-2.
High frequency electrical stimulation reduces neuronal HMGB1 release
Authors: Yang Huan , Datta-Chaudhuri Timir B , George Sam J , Haider Bilal B , Wong Jason , Tracey Kevin J , Chavan Sangeeta S 
Published: January 2023

Abstract
Chronic pain affects the daily lives of millions of Americans and presents an economic burden of $600 Billion per year. Opiods and other pharmaceutical approaches for the management of chronic pain have notorious detremental side effects. Thus, there is a need for understanding of pain for better therapeutics. Neuroinflammation is closely associated with chronic pain and targeting molecular mediators of neuroinflammation may present a novel opportunity for pain treatment. High mobility group box 1 protein (HMGB1), a key mediator of injury- and infection-elicited inflammation, is involved in the pathology of persistent pain. We recently reported that neuronal HMGB1 is required for mediating inflammation and hyperalgesia following nerve injury (Yang H et al. PNAS, 2021). High frenquency (HF) electrical nerve stimulation has achieved clinical success for chronic pain, but the underlying mechanisms are not clear. Here we assess the effects of HF stimulation in modulating HMGB1 release by sensory neurons. Using microelectrode arrays (MEAs) in cultured dorsal root ganglia (DRG) harvested from transgenic mice that express light-sensitive channel rhodopsin in sensory neurons, we observe that light-evoked HMGB1 release from DRGs is significantly reduced with HF stimulation (HMGB1 levels in unstimulated group = 5.3 ± 0.5 ng/ml; in light exposed group = 25.8 ± 6.0 vs. light + HF stimulation = 8.2 ± 2.1* pg/ml, n=6, *: P&lt;0.01). In agreement, HF stimulation (10 min/per day X 3 days) significantly reduces mechanical hyperalgesia and HMGB1 levels in inflamed paws in C57BL/6 mice subjected to sciatic nerve ligation injury. These studies suggest HF stimulation as a novel therapeutic approach to modulate neuronal HMGB1 release for the treatment of pain
Supported by grant from NIH 2R35GM118182-06, to KJT and SSC
Vagus nerve sensory neurons have distinct neural responses to inflammatory mediators
Authors: Huerta Tomas S , Haider Bilal B , Adamovich-Zeitlin Richard , Chavan Sangeeta S , Tracey Kevin J , Chang Eric H 
Published: January 2023

Abstract
Cytokines are secreted signaling proteins that are important mediators of inflammation. While prior work has demonstrated that the level of cytokines can be regulated by nerve stimulation, the role of the nervous system in sensing these immune mediators is still poorly understood. During periods of inflammation, it has been shown that sensory signals travel up the vagus nerve to the brain. However, it is unclear how individual vagal sensory neurons encode specific immune information. Here we use in vivo calcium imaging of the nodose ganglion to monitor neural activity in individual vagus nerve sensory neurons in response to specific cytokines. Using mice that express the calcium indicator GCaMP6f in glutamatergic neurons, we imaged neurons of the nodose ganglion in situ using a 1-photon miniature microscope (Miniscope). During imaging, the proinflammatory cytokines interleukin 1β (IL-1β) and tumor necrosis factor (TNF) were applied directly to the vagus nerve. Fluorescence data was analyzed with a Python-based software CaImAn and a custom pipeline to output the single neuron activity as change in fluorescence: ΔF/F. Our results reveal that specific vagal sensory neurons respond differentially to specific immune mediators. The average amplitude, integral, and delay of TNF-responsive neurons was significantly higher than IL-1β-responsive cells (TNF vs IL-1β, p &lt; 0.01), while having less number of peaks per response (TNF vs IL-1β, p &lt; 0.05). This may suggest different patterns of transient neural activity associated with each particular cytokine. Further investigation into this type of neuro-immune signaling may identify novel neural targets for the treatment of inflammatory disorders.
This study was funded in part by NIH/NIGMS
Structure guided engineering of selective HVEM mutants reveal distinct functions binding to LIGHT and BTLA/CD160
Authors: Chou Ting-Fang , Liu Weifeng , Garrett-Thomson Sarah C , Seo Goo-Young , Fedorov Elena , Ramagopal Udupi A , Bonanno Jeffrey B , Wang Qingyang , Kim Kenneth , Garforth Scott J , Kakugawa Kiyokazu , Cheroutre Hilde , Kronenberg Mitchell , Almo Steven C 
Published: January 2023

Abstract
HVEM is a TNF (tumor necrosis factor) receptor contributing to a broad range of immune functions involving diverse cell types. It interacts with a TNF ligand, LIGHT, and immunoglobulin (Ig) superfamily members BTLA and CD160. Assessing the functional impact of HVEM binding to specific ligands in different settings has been complicated by the multiple interactions of HVEM and HVEM binding partners. To dissect the molecular basis for multiple functions, we determined crystal structures that reveal the distinct HVEM surfaces that engage LIGHT or BTLA/CD160, including the human HVEM–LIGHT–CD160 ternary complex, with HVEM interacting simultaneously with both binding partners. Based on these structures, we generated mouse HVEM mutants that selectively recognized either the TNF or Ig ligands in vitro. Knockin mice expressing these muteins maintain expression of all the proteins in the HVEM network, yet they demonstrate selective functions for LIGHT in the clearance of bacteria in the intestine and for the Ig ligands in the amelioration of liver inflammation.
Supported grants from NIH (S10 OD020068, P30CA023100, P30 DK120515, S10RR027366, U01 AI125955, P01 DK46763), U.S. Department of Energy (DE-AC02-98CH10886, DE-AC02-06CH11357), National Center for Research Resources (P41RR012408), National Institute of General Medical Sciences (P41GM103473), Albert Einstein Cancer Center (P30CA013330), Eli Lilly Company, Albert Einstein Macromolecular Therapeutics Development Facility, Price Family Foundation, Albert Einstein Center for Experimental Therapeutics, Pamela and Edward S. Pantzer, and Academia Sinica, Taiwan.
Regulation of humoral immunity in acute vs. chronic viral infection
Authors: Cooper Lucy , Good-Jacobson Kim L 
Published: January 2023

Abstract
Chronic infectious diseases caused by viral pathogens such as HIV and Hepatitis C constitute a major global health challenge. Chronic viral infections result in poor humoral immune responses, characterized by the delayed generation of neutralizing antibodies and dysregulation of memory B cell formation. Little is known regarding the mechanisms driving dysfunctional B cell memory during chronic viral infection. We used an in vivo mouse model using acute and chronic strains of LCMV, in conjunction with a B cell tetramer specific for the LCMV nucleoprotein, to understand changes that occur in antigen-specific memory B cell formation in acute vs. chronic viral infection. We performed phenotypic analyses of genetically modified mouse strains, BrdU-labelling studies, antiviral treatment experiments and single-cell RNA-sequencing to examine memory B cell formation, phenotype and function in response to acute vs. chronic LCMV infection. Single-cell RNA-sequencing identified key genes downregulated in chronic memory B cells associated with complement-mediated immune functions. We found that T-bet and CXCR3 modulate multiple aspects of the humoral immune response in chronic viral infection, including GC B cell localisation, memory B cell phenotype, and immune complex expansion. Our data provides novel insight into the cellular and molecular mechanisms driving altered humoral immune responses in chronic viral infection.
Testosterone functions as a tumor suppressor in glioblastoma
Authors: Lee Juyeun , Silver Daniel J , Fodor R’ay , Chung Yoon-Mi , Sharifi Nima , Lathia Justin D 
Published: January 2023

Abstract
A sex difference in the incidence and outcome of patients with glioblastoma (GBM), the most common primary malignant brain tumor, has been well established, as male patients experience a worse prognosis. However, the contribution of sex hormones to anti-tumor immunity has not been fully explored. To test this, we orthotopically implanted syngeneic mouse GBM cells into male mice after castration or sham surgery to assess survival differences. Castrated immune-competent mice succumbed to tumors significantly faster than the sham surgery group, and this was rescued by exogenous testosterone administration. In contrast, immune-deficient NSG castrated mice survived longer than sham animals, suggesting a role for immune cells in the survival difference. Flow cytometry analysis of tumor-infiltrating immune cells further showed decreased cytokine production in T cells from castrated mice, while no difference was found in immune cell infiltration. Flank tumor experiments suggested brain-specific regulation of T cell recruitment, as tumor growth was delayed in castrated mice, with higher infiltration of CD8+ T cells but decreased cytokine production. Surprisingly, mass spectrometry of serum revealed upregulated levels of circulating corticosterone in castrated mice regardless of tumor presence, suggesting that castration may induce systemic immunosuppression via upregulation of glucocorticoids. Taken together, our data demonstrate that testosterone regulates T cell immunity, possibly via glucocorticoids, and ultimately affects GBM tumor growth. Given the decreased level of testosterone in aged males with higher risk of GBM, these findings further suggest the potential therapeutic intervention of testosterone in GBM.
Supported by grants from NIH (P01 CA245705)
The role of Piezo1 mechanosensor in osteoclastogenesis from monocytes
Authors: Rawas-Qalaji Mohamad , Shindo Satoru , Nakamura Shin , Kawai Toshihisa 
Published: January 2023

Abstract

Objectives
Interrelations between bone and the immune system are involved in both homeostatic and pathogenic bone remodeling processes. Although emerging evidence supports that mechanosensitive Ca2+-channel Piezo1 plays a pivotal role in osteoblast-mediated bone regeneration, possible engagement of Piezo1 in the differentiation of osteoclasts from monocytes and resulting bone resorption is largely unknown. In the present study, we sought to establish the role of Piezo1 in RANKL-stimulated monocytes for their differentiation to osteoclasts in vitro.

Methods
Mouse RAW 264.7 monocyte cell line or bone marrow derived mononuclear cells (BMMCs) were stimulated with M-CSF and/or RANKL, in the presence or absence of Yoda1 (Piezo1 activator) or with or without hydrostatic pressure (HP). Ca2+ influx assay was employed to confirm Piezo1 channel activity. To evaluate the level of osteoclastogenesis and bone-resorptive function by mature osteoclasts, TRAP staining and pit formation assays were performed. To monitor the expressions of mRNAs for Piezo1 as well as OC-STAMP, TRAP (ACP5) and MMP9, qPCR were employed.

Results
Yoda1-mediated Ca2+ influx was significantly elevated in RANKL BMMCs and RAW264.7 cells, while significantly suppressing the expressions of osteoclastogenesis-related genes, OC-STAMP, MMP9 and ACP5. Moreover, the number and size of multi-nucleated TRAP-positive cells as well as formation of resorption pits were significantly down-regulated by the treatment with Yoda1 as well as HP.

Conclusions
The present study suggests that mechanosensitive Piezo1 channel expressed on monocytes may play a gatekeeper role in down-modulating over-activation of osteoclastogenesis and function to resorb bone.
Supported by NIH NIDCR grants, DE-027851, DE-028715 and DE-331851