bluuebunny/crossref_metadata_2025_split huggingface dataset:
p38 MAP kinase signaling in microglia plays a sex-specific protective role in CNS autoimmunity
Authors: Krementsov Dimitry N , McGill Mahalia 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an autoimmune demyelinating disease of the central nervous system, representing the leading cause of non-traumatic neurologic disease in young adults. This disease is 3 times more common in women, but the underlying mechanisms remain unknown. MS is initiated by autoreactive T helper cells, but CNS-infiltrating inflammatory myeloid cells are the main effector cells driving disease pathology. The role of CNS-resident macrophage-like microglial cells is unclear. We have previously shown that genetic ablation of p38α MAP kinase (p38) in the myeloid lineage is protective in the autoimmune model of MS, experimental autoimmune encephalomyelitis (EAE). Strikingly, protection was seen in female mice, and not males. Here, we sought to precisely define the mechanisms responsible. To this end, we used multiple genetic approaches and bone marrow chimeras to ablate p38 in microglial cells, peripheral myeloid cells, or both. Deletion of p38 in both cell types recapitulated the previous sex difference, with reduced EAE severity in females. Unexpectedly, deletion of p38 in the periphery was protective in both sexes, while deletion of p38 in microglia exacerbated EAE only in males. Transcriptional profiling of microglia isolated from the CNS at peak of EAE revealed male-specific p38-dependent transcriptional modules with anti-inflammatory/tissue reparatory functionality. Taken together, these results reveal the presence of a p38-dependent male-specific molecular pathway in microglia that is protective in CNS autoimmunity, suggesting that autoimmunity in males and females may be driven by distinct cellular and molecular pathways, thus informing the design of novel future sex-specific therapeutic approaches.
Developmental exposure to environmental toxicant trichloroethylene alters DNA methylation in polycomb protein binding regions in effector/memory CD4 + T cells from autoimmune-prone mice
Authors: Blossom Sarah J , Byrum Stephanie , Washam Charity L , Patterson John D. , Vyas Kanan K , Gilbert Kathleen M. 
Published: January 2023

Abstract
Trichloroethylene (TCE) is an industrial solvent and drinking water pollutant associated with autoimmunity in human populations. Previous studies in autoimmune-prone MRL+/+ mice revealed autoimmune pathology and CD4 + T cell autoreactivity were more robust when exposure occurred during development rather than adulthood. To determine whether these effects were linked to epigenetic changes, mice were exposed to vehicle control or TCE in the drinking water beginning at gestation until 37 weeks of age. In some mice, TCE was removed from drinking water at 22 weeks of age. Whole genome reduced representation bisulfite sequencing (RRBS) was used to compare methylation of CpG sites in autosomal chromosomes in effector/memory CD4+ T cells. Few significant changes occurred in mice when TCE was removed from drinking water. In contrast, continuous developmental TCE exposure revealed differential methylation of 252 CpG sites relative to controls. When CpG sites were overlapped with the Open Regulatory Annotation database (ORegAnno), continuous TCE treatment resulted in 130 differentially methylated regions (DMRs) that included 12 unique transcription factors. Interestingly, 80% of DMRs occurred in areas known to bind Polycomb group (PcG) proteins that form the polycomb-repressive complex 2 (PRC2) in mammalian cells, namely, SUZ12, EZH2, JARID2, and MTF2. Pathway analysis of the DMRs indicated that TCE primarily altered the methylation of genes asociated with regulation of cellular metabolism and cell signaling pathways. In summary, TCE differentially methylated binding sites of PcG proteins in effector/memory CD4+ cells, and point toward a novel mechanism by which TCE could modulate CD4 + T cell function and promote autoimmunity.
PDLIM4: Target of JMJD3 regulates CD4 T cell Trafficking by connecting S1P1 and cytoskeleton
Authors: Fu Chuntang , Li Qingtian , Zou Jia , Xing Changsheng , Yu Jiaming , Chu Junjun , Yin Bingnan , Luo Mei , Wang Rongfu 
Published: January 2023

Abstract
In 2018 AAI annual meeting, we presented our discover that JMJD3 was bound to the promoter and gene body regions of Pdlim4 gene and it regulated PDLIM4 expression by interacting with zinc finger transcription factor KLF2. PDLIM4 as an filamentous actin (F-actin) binding protein, was involved in the T cell trafficking. However, the role and mechanisms of PDLIM4 in T cell trafficking remain unclear. In this abstract, we showed that PDLIM4 as an adaptor protein connecting S1P1 and filamentous actin (F-actin) was a key regulator of T cell trafficking. Our studies indicated that PDLIM4 interacted with S1P1 by N terminal PDZ domain. The PLDIM4 C terminal LIM domain anchored the complex to filamentous actin (F-actin). Our findings identified Pdlim4 as a novel Jmjd3 target gene that affects T-cell trafficking by cooperating with S1P1 and provided insights into the molecular mechanisms of which Jmjd3 regulates genes participating in thymic emigration.
Role of CXCR3 in natural killer cell migration to melanoma cells
Authors: Park Eun Jae , Kang Yu Jeong , Lee Hong Kyung , Kim Hyung Sook , Hong Jin Tae , Kim Youngsoo , Han Sang-Bae 
Published: January 2023

Abstract
Natural killer (NK) cell recruitment to solid tumors is essential for direct contact between NK and cancer cells and is regulated by chemokine networks. Tumor-derived CXCL10 increases the infiltration of NK cells into solid tumors. In a xenograft mouse model, adoptively transferred human NK cells efficiently infiltrate CXCL10-transduced tumors and enhance regression of tumor growth compared with mock-transduced tumors. Although this study showed that NK cells homed to solid tumor beds in a CXCL10-depedent manner, several questions remain. How do NK cells find cancer cells after extravasation? Which chemokines do B16F10 cells produce? Which chemokine receptors do NK cells use? How do cancer cell-derived chemokines affect NK cell motility? Do these chemokines directly affect NK cell cytotoxicity? To address these issues, we examined the migration dynamics of NK and cancer cells using time-lapse imaging. Although naïve B16F10 cancer cells produce low levels of chemokines, IFN-g-treated B16F10 cells secreted high levels of CXCL10, low levels of CCL5, but did not secrete CCL2, CCL7, or CXCL12. Wild-type NK cells migrated well toward cancer cells and killed them, whereas NK cells deficient in CXCR3 did not. CXCR3-deficient NK cells also showed slower migration speed than did wild-type NK cells. Taken together, our data show that NK cells find cancer cells, at least in part, by sensing CXCL10 produced by cancer cells and suggest that a strategy to increase CXCL10 secretion by cancer cells may improve the efficacy of NK cell–based immunotherapy.
PIEZO1 forms an adhesive-mechanosensitive complex with activated LFA-1 on T lymphocytes
Authors: Aguilar Alicia Virginia , Benson Bryan , Rathkey Joseph , Correa Luis , Li Lucy , Myers Jay , Tomalka Jeffrey , Kennelley Hannah , Abbott Derek , Ransohoff Richard , Huang Alex 
Published: January 2023

Abstract
In order to respond to infection, injury and stress, T lymphocytes must successfully migrate from the bloodstream into inflamed tissue in a process called the leukocyte adhesion cascade. Of these steps, blockade of adhesion has shown clear clinical benefit in autoimmunity, and the step of intraluminal crawling is of great potential clinical importance. In order to coordinate adhesion and crawling while in circulation, leukocytes must respond appropriately to complex hemodynamic forces, including shear stresses and erythrocyte driven margination against the venule walls. Therefore, we hypothesized that the mechanosensitive calcium channel PIEZO1, which is preferentially expressed in T lymphocytes, mediates T lymphocyte responses to force during adhesion and crawling. To test this hypothesis, we genetically ablated PIEZO1 in primary T lymphocytes from healthy human donors using CRISPR/Cas9, and observed crawling ability in vitro. In this assay, PIEZO1 knockout cells exhibited decreased crawling and disrupted morphology. Moreover, we performed immunofluorescence of PIEZO1 on chemokine-activated crawling T lymphocytes, which demonstrated that PIEZO1 redistributes to the contact zone of crawling cells in a pattern reminiscent of a high-affinity LFA-1 focal zone. Additionally, PIEZO1 colocalizes specifically with high affinity LFA-1. Using co-immunoprecipitation indicated that PIEZO1 preferentially associates with the alpha integrin subunit of LFA-1 in its active form, through a conserved amphipathic eight amino acid motif. Taken together, our data suggest that PIEZO1 contributes to coordination of T lymphocyte crawling via interaction with integrins, modulating the affinity state and turnover of LFA-1.
Dendritic Cell-Induced Dissemination of Mycobacterium tuberculosis into the Central Nervous System
Authors: Gilpin Trey E , Walter Fruzsina R , Herbath Melinda , Sandor Matyas , Fabry Zsuzsanna 
Published: January 2023

Abstract
Central nervous system tuberculosis (CNSTB) is the most serious manifestation of extrapulmonary tuberculosis infections representing 5–10% of all cases. Knowledge of CNSTB stems from observations made in 1933 by pathologists Rich and McCordock, who found caseating foci in the brain parenchyma or meninges that rupture into the subarachnoid space leading to diffuse meningeal infection. However, the mechanism of how immobile Mycobacterium tuberculosis, the causative agent of TB crosses the highly regulated blood-brain barrier (BBB) and enters the CNS forming these foci and infection is unknown. We have developed a novel in vitro BBB model that combines Mycobacteria-infected DCs, PBMCs, and primary mouse brain endothelial cells to observe early cellular interaction and aggregate formation at the BBB similar to previous observations by Rich and McCordock. With this model we observed Mycobacteria impedes migration of DCs but increases DC initiation of aggregation on a primary mouse endothelial BBB with a cellular composition similar to pulmonary granulomas. Aggregates induce an inflammatory response by increasing ICAM-1 and VCAM-1 expression and dysregulation of tight junction proteins and mitochondria suggesting barrier damage possibly due to increased iNOS expression observed during within aggregates. Mycobacteria-infected DC transmigration is increased in the presence of aggregation. Previous research from our lab and others have shown DC transmigration across an in vitro BBB rely on MMP-9 secretion while TNF-alpha is necessary for pulmonary granuloma formation and Mtb infection control. Here we will show the role iNOS, MMP-9 and TNF-alpha play in DC-mediated dissemination of Mycobacterium across the BBB.
Mass-cytometry reveals global immune remodeling with multi-lineage hypersensitivity to Type I Interferon in Down syndrome
Authors: Waugh Katherine A. , Pandey Ahwan , Jordan Kimberly R , Smith Keith P , Granrath Ross E , Khanal Santosh , Butcher Eric T , Rachubinski Angela L , McWilliams Jennifer A , Araya Paula , Minter Ross , Dimasi Tiana , Colvin Kelley L. , Baturin Dmitry , Pham Andrew T , Galbraith Matthew D , Yeager Michael E. , Sullivan Kelly D , Porter Christopher C , Hsieh Elena W , Espinosa Joaquin M 
Published: January 2023

Abstract
People with Down syndrome (DS), the genetic condition caused by trisomy of chromosome 21 (chr21), have an altered disease spectrum. For example, adults with DS are largely protected from solid tumors while predisposed to various autoimmune disorders, early-onset Alzheimer’s disease, and severe respiratory infection. While the immune system is known to play a key role in these comorbidities among the disomic population, there is a dearth of immune studies in adults with DS. Here, we applied the single-cell, systems-level approach of mass-cytometry, or Cytometry by Time-Of-Flight (CyTOF), to broadly define phenotypic and functional alterations in immune homeostasis in adults with DS that are relevant to the potentiation of comorbidities associated with T21. We developed an innovative strategy to analyze CyTOF data, by incorporating techniques commonly used in genomics and even topography to eliminate batch bias and efficiently resolve ~100 immune cell types. This approach revealed that adults with DS have global immune dysregulation reminiscent of various inflammatory and autoimmune states enriched among older individuals of the disomic population. Upon stimulation directly ex vivo with Type I Interferon (IFN) and simultaneous detection of diverse phosphorylation events across the lymphoid and myeloid lineages, all immune subsets from individuals with T21 were hyper-responsive, but the degree of response and signaling pathways used varied among specific cell types. Notably, 4 of the 6 IFN receptor subunits for Types I, II, and III IFNs are encoded on chr21. Altogether, these results establish a critical foundation to pursue immune dysregulation and IFN hyperactivity as a driver of the altered disease spectrum in adults with DS.
Interleukin-37 Promotes Colitis-associated Carcinogenesis (CAC) via SIGIRR-mediated Cytotoxic T Cells Exhaustion
Authors: Li Jiong , Teng Xiu , Wei Xiaoqiong , Wang Zhen , Zeng FanLian , Zhou Hong , Zheng Huaping 
Published: January 2023

Abstract
Interleukin-37 (IL-37), a ligand of the IL-1 cytokine family, was identified as a fundamental inhibitor of innate immunity, having emerged as a dual-function cytokine with extracellular (ligation with receptor SIGIRR) and intra-(interaction with smad3) properties for regulating inflammatory response. However, the role and molecular mechanism of IL-37 in CAC has been rarely reported. IL-37 expression was highly expressed in human colorectal cancer tissues and predicted poor prognosis. Indeed, mice with transgenic expression of IL-37 (IL-37tg mice) were highly susceptible to colorectal cancer induced by AOM/DSS and suffered from dramatically increased tumor burdens in the colon. Nevertheless, aggravated epithelial disruption in IL-37tg mice was observed during DSS-induced injury, which was dependent on the presence of commensal bacteria through MAPK and mTOR pathways. Plus, IL-37 did not affect growth or enhance mutagenesis of transformed colonic epithelia cells. These results demonstrated that decreased proliferation of epithelia cells in response to DSS injury in CAC model is unlikely to be responsible for susceptibility in IL-37tg mice. Significant deficiency and dysfunction of cytotoxic T cells was consistently found in colonic lamina propria in IL-37 tg mice. This exhaustion led by IL-37 was dependent on checkpoint SIGIRR-mediated TIR signaling blocking, independent of TGF-β/smad3 signaling agonizing or antagonizing, and not caused by common immune-checkpoints. Our findings highlight a role for IL-37 in harnessing antitumor immunity by inactivation of cytotoxic T cells and establish new inhibitory checkpoint IL-37/SIGIRR in the cancer-immunity cycle as therapeutic targets in colorectal cancer.
Human IL10 promoter haplotypes confer differential outcomes to experimental colitis in humanized BAC mice
Authors: Vacharathit Vimvara , Ma Jinxia , Gunasekera Dilini C. , Shah Palak , Sheh Alexander , Brayton Cory F. , Whary Mark T. , Fox James G. , Bream Jay H. 
Published: January 2023

Abstract
Most genetic variants associated with complex diseases map to non-coding regions of the genome, but determining their phenotypic consequences remains challenging. We devised a bacterial artificial chromosome (BAC) transgenic approach to model the effects of human non-coding single nucleotide polymorphisms (ncSNPs) on complex disease outcomes. As proof-of-concept, we focus on human IL10, a potent anti-inflammatory cytokine and key mediator of gut homeostasis. ncSNPs in the IL10 locus have been strongly associated with a variety of disease risks, including colitis susceptibility, but their biological functions are unclear. We created two genetically humanized IL10BAC transgenic mouse lines based on two common and well-defined −1082/−819/−592 IL10 promoter haplotypes (“ATA” and “GCC”) on a mouse Il10-deficient background. Although both IL10BAC strains were protected from spontaneous colitis, only ATA-bearing mice were resistant to dextran sulfate sodium (DSS)-induced acute colitis, while GCC and WT mice had high rates of morbidity and mortality. We found that ATAs had different kinetics of cell-type-specific IL-10 production pre- and post-DSS treatment compared to GCCs and WTs, which influenced the expression profiles of other cytokines and led to distinct immunophenotypes. The skewed responses culminated in the inability of GCCs and WTs to control DSS-induced bacteremia, resulting in death. Our data suggest that human IL10 ncSNPs impart allele-specific differences in spatiotemporal regulation of IL-10 expression, leading to differential outcomes to experimental colitis. Humanized BAC mice may serve as an important tool in modeling the potential impact of human non-coding variants on complex biological outcomes.
Detecting IL-1β release with a rapid, homogeneous, bioluminescent immunoassay
Authors: O’Brien Martha A. , Lindstedt Anders , Lazar Dan , Cali James 
Published: January 2023

Abstract
IL-1β is an important inflammatory cytokine of the innate immune system, released from monocytes, macrophages, and other cell types in response to a variety of pathogens and danger signals consequent to inflammasome activation. To enable direct measurement of released IL-1β from cells (no transfers or wash steps), two anti-human IL-1β antibodies were respectively labeled with complementary subunits, SmBiT and LgBiT, of the NanoLuc® Binary Technology. When the labeled antibodies bind to released IL-1β, the complementary subunits are brought into proximity, thereby reconstituting a bright luciferase that generates light when the furimazine substrate is added. Luminescence is monitored with a standard plate-reading luminometer. This novel assay effectively detected released IL-1β from differentiated human monocytic THP-1 cells and human peripheral blood mononuclear cells (PBMCs) stimulated with several TLR agonists, including endotoxin and non-endotoxin pyrogens, and could enable a simplified method for a Monocyte Activation Test for pyrogen detection. Released IL-1β can be detected directly in cell culture assay wells or in a secondary plate with transferred culture medium. Benchmarking assay performance to a standard IL-1β ELISA verified the accuracy of this more rapid, homogeneous IL-1β assay method. Of note, this IL-1β assay can be multiplexed with a homogeneous caspase-1 activity assay, providing complementary high throughput methods to identify modulators of inflammasome activation. MCC950, an NLRP3 inflammasome inhibitor, was shown to inhibit IL-1β release from treated THP-1 cells and PBMCs. Application of this homogeneous immunoassay approach to other inflammatory cytokines is currently being explored.
A circulating NAD biosynthetic enzyme is a novel modulator of inflammation
Authors: Manago’ Antonella , Audrito Valentina , Mazzola Francesca , Sorci Leonardo , Gaudino Federica , Gizzi Katiuscia , Incarnato Danny , Minazzato Gabriele , Ianniello Alice , Varriale Antonio , D’Auria Sabato , Mengozzi Giulio , Politano Gianfranco , Oliviero Salvatore , Raffaelli Nadia , Deaglio Silvia 
Published: January 2023

Abstract
Damage-associated molecular patterns (DAMPs) are molecules that can be actively or passively released by injured tissues and that activate the immune system. Here we show for the first time that the NAD biosynthetic enzyme nicotinate phosphoribosyltransferase (NAPRT), the rate-limiting enzymes in the intracellular synthesis of NAD from nicotinic acid, is physiologically present in human sera, where it acts as a novel DAMP. We detected NAPRT in plasma/sera from human donors by antibody-mediated luminex assays and mass spectrometry. Exposure of human and mouse macrophages to NAPRT triggers activation of ERK1/2, phosphorylation of IKKα/β and nuclear translocation of the p65 subunit of the NF-kB complex, with synthesis and secretion of inflammatory cytokines, including IL-1β, IL-8, TNFα and CCL3. Furthermore, NAPRT enhances monocyte differentiation into macrophages, by inducing macrophage colony-stimulating factor. These effects are independent of the NAPRT catalytic activity, but rely on the protein’s binding to TLR4, as demonstrated by showing direct in vitro interaction and by the in vivo lack of NAPRT effects in TLR4−/− macrophages. In line with the finding that NAPRT mediates endotoxin tolerance, sera from patients with sepsis or septic shock contain the highest levels of circulating NAPRT, compared to other chronic inflammatory conditions, including cancer. Importantly, patients with serum NAPRT &gt;15 ng/ml are characterized by a worse clinical outcome, compared to the counterparts.
Together, these data identify NAPRT as a novel endogenous ligand for TLR4 and a critical mediator of inflammation.
Peripheral blood CD115 (CSF-1R) expression is anticoagulant- and time-dependent
Authors: Acuff Nicole V , Divekar Anagha A , Yang Xifeng 
Published: January 2023

Abstract
Human CD115, also known as CSF-1R, is highly expressed by monocytes, macrophages, and dendritic cells. Here we show that detection of human CD115 on peripheral blood monocytes is dependent on both the anticoagulant and the storage time before sample processing. Commercially available CD115 antibody, clone 9-4D2-1E4, shows bright staining on blood samples collected in EDTA and citrate based anticoagulants and a dimmer staining intensity in blood collected in heparin. Clone 9-4D2-1E4 is not compatible with Cyto-Chex™ tubes. Consistent with published reports in mice, human CD115 signal is reduced within hours of collection. CD115 expression is further reduced after overnight storage at ambient temperature and is completely lost two days post collection. Reduced CD115 expression over time is most noticeable in whole blood collected in heparin. These results demonstrate that anti-coagulant and storage time are important considerations for current research projects and clinical trials targeting CD115 (CSF-1R).
CD301b+ SIRPα+ dendritic cells play a non-redundant role in inducing clonal deletion in the thymus medulla
Authors: Breed Elise R , Hogquist Kristin A 
Published: January 2023

Abstract
The thymus contains an assortment of antigen presenting cells (APC) that have distinct locations and antigen processing capabilities. This suggests that they play non-redundant roles in mediating thymocyte selection. Many APC subsets were shown to be capable of driving clonal deletion, however these studies relied primarily on T cell receptor transgenic models. We therefore still do not understand the extent to which distinct APC contribute to clonal deletion in the polyclonal repertoire. Here, we assessed the contribution of different APC subsets to clonal deletion using a cleaved caspase 3-based assay paired with cell type ablation or deficiency. Total deletion frequencies were not altered in the absence of B cells, pDC, or XCR1+ cDC1. In an effort to eliminate SIRPα+ cDC2, we discovered that a substantial proportion of thymic SIRPα+ DC express the lectin CD301b. CD301b+ DCs were localized exclusively within the thymus medulla and depended on IL-4R□. Deficiency of these IL-4/IL-13 signaled DC resulted in a measurable reduction in clonal deletion. These data suggest that CD301b+ SIRPα+ DC represent a distinct population of antigen presenting cells within the thymus and that they are essential for non-redundant deletional tolerance in the polyclonal repertoire.