bluuebunny/crossref_metadata_2025_split huggingface dataset:
Thpok promotes CD4+-lineage differentiation by recruiting the NuRD chromatin remodeling complex
Authors: Gao Yayi , Zamisch Monica , Vacchio Melanie S , Paine Elliott , Lyons Gaelyn , Ciucci Thomas , Chopp Laura , Xiao Qi , Vinson Charles , Jenkins Lisa , Bosselut Rémy 
Published: February 2023

Abstract
T cell precursors develop into helper CD4 or cytotoxic CD8 T cells in thymus. Thpok, a BTB/POZ family transcription factor, enforces MHC II restricted thymocytes commitment to the CD4 lineage, while Runx3 protein drives MHC I restricted thymocytes to the CD8 lineage. However, the molecular mechanisms underlying Thpok functions during CD4 T cell development are not understood. Replacing the BTB domain from Thpok with that from Bcl6, another BTB/POZ zinc finger transcription factor that recruits NCOR-family co-repressors through its BTB domain, abrogated CD4 T cell development in thymus; this suggests a unique feature associated with Thpok BTB domain. Here, using mass-spectrometry, we identified nucleosome remodeling and deacetylase (NuRD) complex as a novel Thpok cofactor. We demonstrated that the Thpok BTB domain is essential for NuRD recruitment. Moreover, we identified BTB domain amino acid residues critical for NuRD complex interaction, and showed that mutation of these Thpok residues disrupts CD4 T cell development in vivo. Reciprocally, a fusion protein reconstituting NuRD binding to a BTB-less version of Thpok (which cannot bind NuRD) restored CD4 T cells in vivo. In addition, we found that CD4-lineage thymocytes expressing that fusion protein have a transcriptome similar to that of thymocytes expressing wild type Thpok. Finally, we found that NuRD binding is required for Thpok to antagonize Runx3 expression in thymocytes. Thus, our results demonstrate that NuRD recruitment is both necessary and sufficient for the functions of the Thpok BTB domain in CD4 T cell development and it is also required to antagonize Runx3-dependent differentiation towards CD8 lineage.
Pathogenic effector TH2 cells discriminate between allergic and eosinophilic inflammation in food allergy and eosinophilic gastrointestinal disease
Authors: Makiya Michelle A , Frischmeyer-Guerrerio Pamela A , Khoury Paneez , Klion Amy D. , Kuang Fei Li 
Published: February 2023

Abstract
IgE-mediated food allergies (FA) and delayed food-triggered (FT) gastrointestinal tissue eosinophilia (EGID) have distinct clinical manifestations and are treated differently. Pathogenic effector TH2 (peTH2) cells are implicated in inappropriate TH2 responses in both of these diseases. Furthermore, EGID patients can have concomitant FA, and some EGID patients do not report any food triggers. To better understand the role of peTH2 cells in the context of FA and EGID and the overlap between these two conditions, peTH2 cells (CD3+CD4+CD27-CD49d+CRTH2+CD161+) from purified peripheral blood mononuclear cells (PBMC) were profiled by intracellular cytokine flow cytometry in these deeply-characterized patient cohorts: patients with FA alone (FA, n=5), EGID with FA and FT (EGID+FA+FT, n=7), EGID with FT (EGID+FT, n=7) and EGID without FA or FT (EGID, n=9). Patients with any food allergy or food trigger (FA, EGID+FA+FT, and EGID+FT) have increased peTH2 cells compared to healthy volunteers (HV). Interestingly, a greater percentage of peTH2 cells in patients with food triggers (EGID+FA+FT, EGID+FT) exhibit a trend toward IL5+IL13+ double positivity (P = 0.0556) than those from patients without food triggers (FA, EGID). In contrast, peTH2 cells from patients with FA had a greater percentage that were IL17A+ (P=0.009) or IFNγ+ (P=0.0444) than patients with eosinophilic disease (EGID+FA+FT, EGID+FT, EGID). Our studies indicate that while elevated levels of peTH2 cells are indicative of allergic inflammation of both immediate hypersensitivity and eosinophilic disease, the cytokine profile of each disorder may be distinct, where IL17A+ and IFNγ+ predominate in those without eosinophilic disease.
Plasma cells Dynamics in the Bone Marrow Niche
Authors: Jing Zhixin , Benet Zachary L , Fooksman David R 
Published: February 2023

Abstract
Plasma cells (PCs) can become long-lived with age to maintain prophylactic antibody titer for decades in the bone marrow (BM) microenvironment that are enriched in supporting chemokines and pro-survival cytokines. It has been that PCs are sessile in the BM to receive various pro-survival signals, but how they access these signals in the dynamic BM microenvironment remains unknown. Here by establishing long-term intravital imaging of the mouse tibial BM, we show that PCs are overall motile within the BM parenchyma with unique intermittent and heterogenous motility over time. Their motility is reduced when they are in clusters or as they enter a cluster. PC cluster formation and motility requires the pro-survival cytokine APRIL. PCs motility also requires the chemokine receptor CXCR4 and its ligand CXCL12 that are known important for PC homing to BM. CXCR4/CXCL12 signaling-triggered integrin VLA-4 activation negatively regulates PC motility in the BM. Notably, not only do PCs migrate within the BM parenchyma, but they can also egress and recirculate under steady state condition or CXCR4 and VLA4 inhibition. PC motility, clustering, and recirculation are all increased with mouse age. Thus, PC motility and cluster formation underlie a dynamic BM survival niche, contributing to PC longevity and function.
Plasmacytoid Dendritic Cells Regulate Th17 Activation in Diabetic Wound CD4+ T-cells
Authors: Audu Christopher O , Wolf-Fortune Sonya , Melvin William James , Davis Frank , Sharma Sriganesh B. , Mangum Kevin , Barrett Emily , Joshi Amrita , Obi Andrea T. , Moore Bethany B , Gallagher Katherine A. 
Published: February 2023

Abstract
Introduction: CD4+ T-cell activation is vital for normal wound repair but the factors that control T-cell activation in wounds in vivo are not clear. Our group and others have found increased Th17 activation in diabetic wounds resulting in increased IL-17a and pathologic inflammation that prevents tissue repair. Plasmacytoid dendritic cells (pDC) are antigen presenting cells that are present in early diabetic wound tissue and may play a key role in modulating CD4+ T-cell phenotype. Hence, we hypothesized that diabetic pDCs may influence wound CD4+ T-cells towards Th17 T-cell expansion.

Methods:
Wild type C57BL/6 mice were fed normal chow diet (13.5% kcal fat; LabDiet) or high fat diet chow (60% kcal fat; Research Diets) for 12–14 weeks to generate the diet-induced obesity (DIO) model of glucose intolerance/insulin resistance. These mice were subsequently wounded, and wound plasmacytoid dendritic cells harvested on day 1 and day 3. These cells were co-cultured with naïve CD4+ T-cells for 3 days, after which T-cell phenotype was determined by flow cytometry. Additionally, pDC in wounds 1-day post wounding were examined by quantitative PCR for cytokine production.

Results:
Following exposure to DIO pDCs, wild type activated CD4+ T-cells were activated towards a Th17 phenotype via significant increases in TGFb CD4+ T-cell activation and may act to increase inflammation in diabetic wounds.
BCG-Infected Dendritic Cells Induce TNF-alpha Dependent Cell Cluster Formation That Damage theBlood Brain Barrier
Authors: Gilpin Trey E , Walter Fruzsina , Herbath Melinda , Sandor Matyas , Fabry Zsuzsanna 
Published: February 2023

Abstract
Central nervous system tuberculosis (CNSTB) is the most serious manifestation of extrapulmonary tuberculosis infections representing 5–10% of all cases. How Mycobacterium tuberculosis (Mtb) crosses the blood-brain barrier (BBB) and enters the CNS is not well understood. We applied a novel in vitro model of BBB model that consists of primary brain astrocytes and microvessel endothelial cells to test the mechanisms how mycobacterium-infected DCs interact with peripheral blood mononuclear cells (PBMC) and affect the BBB. We show that mycobacterium hinders DC migration, and infected DCs adhere to brain microvessel endothelial cells and induce PBMC cell cluster formations. Infected DC-induced cell clusters contain 24% CD11c+, 57%CD11b+, and 16% CD4+ cells. PBMC and infected DC clusters strongly adhered to the in vitro BBB and over time induced cluster associated damage (CAD). CAD correlated with mitochondrial disorganization and the degradation of tight junction proteins ZO-1 and claudin-5, and upregulation of cellular adhesion molecules VCAM-1 and ICAM-1 on endothelial cells. Anti-TNF-a inhibition reduced cell cluster formation as well as ICAM-1 expression on brain microvessel endothelial cells and mitigated CAD. This research leads to better knowledge in understanding how mycobacteria infected cells affect the BBB that could lead to better therapeutic treatments for mycobacterial infection of the CNS.
Astrocyte TAK1-mediated production of chemotactic cues are necessary for the localization of CD4+T cell in gray matter in an EAE model with severe neurodegeneration.
Authors: Khaw Yee Ming , Steelman Andrew J. , Inoue Makoto 
Published: February 2023

Abstract
Multiple sclerosis (MS) is a chronic neurological disease of the central nervous system that is driven by peripheral immune cell infiltration and glial cell activation. The pathological hallmark of MS is primary demyelination, but recent evidence suggests gray matter neuronal damage is a major component of the irreversible nature of the disease. While T cells are found in both gray and white matter of MS tissue, they are typically confined to the white matter of the most commonly used mouse model of MS, experimental autoimmune encephalitis (EAE). Thus, genesis of T cells in the gray matter during MS pathogenesis is poorly understood. In this study, we used a modified EAE model (Type-B EAE) that displays severe neuronal damage without disease remission to investigate the role of T cells in neurodegeneration. Using immunohistochemistry and confocal imaging, we show that CD4+ T cells migrate to the spinal cord gray matter with selective preference for the ventral horns. Compared to CD4+ T cells in white matter, gray matter-infiltrated CD4+ T cells were mostly immobile (indicative of cell–cell interactions) and can be seen interacting directly with neuronal bodies. T cell-specific deletion of CXCR2 significantly decreased CD4+ T cell infiltration into gray matter in Type-B EAE mice. Inhibition of astrocyte chemokine production such as CXCL1, which was heightened in gray matter of Type-B EAE mice, by astrocyte-specific deletion of TAK1 successfully prevented T cell migration into spinal cord gray matter and rescued neuronal damage and motor dysfunction in Type-B EAE mice. This is the first report report that identifies the mechanism by which encephalomyelitic CD4+ T cells migrate to spinal cord gray matter, which correlates with neuronal damage.
Hypoxia inducible factor-1a drives metabolic adaptation of natural killer cells during pathogen infection
Authors: Victorino Francisco , Bigley Tarin , Park Eugene , Yao Conghui , Benoit Jeanne , Yang Li-Ping , Plougastel-Douglas Beatrice , Piersma Sytse , Lauron Elvin , Davidson Rebecca , Patti Gary , Yokoyama Wayne 
Published: February 2023

Abstract
Natural killer (NK) cells provide an early response to pathogen infection, limiting host morbidity and lethality. This response requires NK cells to adapt due to the energy demands of activation and biosynthesis. We found that NK cells upregulate the metabolic adaptor hypoxia inducible factor-1a (HIF-1a) during murine cytomegalovirus (MCMV) infection in vivo. Specific deletion of HIF-1a in NK cells impaired their ability to reduce viral load and protect against morbidity. While HIF-1a-deficient NK cells had no defects in effector functions, their numbers were significantly reduced during early responses and there was blunted expansion of Ly49H+ NK cells. However, NK cell proliferation per se was not impaired during in vivo infection and in vitro cytokine stimulation. Instead, we found that the absence of HIF-1a impaired glucose metabolism and up-regulated the pro-apoptotic protein Bim during cytokine stimulation in vitro. Furthermore, during MCMV infection in vivo HIF1a-deficient NK cells upregulated Bim and had increased caspase activity. Thus, HIF-1a normally drives glucose metabolism and represses Bim expression during pathogen infection to sustain NK cell numbers for an optimal response.
Cell death effectors are recruited to Toxoplasma gondii parasite vacuoles targeted by interferon-inducible GTPases in infected dendritic cells
Authors: Zhao Xiaoyu , Yin Bocheng , Ewald Sarah E. 
Published: February 2023

Abstract
Toxoplasma gondii is an obligate intracellular pathogen whose ability to grow and evade cell autonomous immunity depends on the parasitophorous vacuole membrane (PVM). The PVM forms from the host plasma membrane during invasion and is maintained by parasite effectors. Interferon-inducible GTPases (IIGs) are central in parasite clearance, however, the precise mechanism linking IIG attack of the PVM, parasite killing and inflammatory host cell death is not clear. To identify host and parasite proteins recruited to the PVM, we used Automated Spatially Targeted Optical Micro Proteomics (AutoSTOMP), a novel discovery technique that uses a confocal microscope to visualize and photo-label proteins for identification by LC-MS. Using IFNγ to induce IIG attack of the PVM and the TLR2 ligand Pam3CSK4 to upregulate inflammasome components we examined protein recruitment to the PVM in BMDCs. AutoSTOMP identified over 300 proteins differentially enriched near the PVM of unprimed, TLR2, IFNγ and IFNγ+TLR2 treated BMDCs. To identify candidate regulators of parasite clearance, we selectively purified proteins on the PVMs coated with IIG protein GBP2. We identified the inflammasome components caspase-1 and ASC as well as the apoptotic caspases-3 and -6 on GBP2-coated PVM. This was consistent with the observation that IFNγ priming was sufficient for host cell death and parasite restriction. BMDC death was partially dependent on the Aim2, ASC, Caspase1/11 and gasdermin D, however, blocking the inflammasome didn’t impair parasite restriction. By contrast, iNOS inhibitor, was sufficient to partially rescue parasite growth. In summary, we report a new tool to study the PVM proteome and identify regulators of parasite clearance and host cell death.
Mechanistic investigations of Granzyme A-mediated TB protective effects
Authors: Rasi Valerio , Xia Mei , Wood David , Eickhoff Christopher , Hoft Daniel F 
Published: February 2023

Abstract
One fourth of the world population is infected with Mycobacterium tuberculosis (Mtb), the pathogen that causes Tuberculosis (TB). Our lab has identified γδ T cells that secrete Granzyme A (GzmA) with TB protective effects. To elucidate the mechanism(s) involved in GzmA-mediated mycobacterial pathogen inhibition using a relevant in vitro human system. This GzmA was utilized to treat mycobacteria-infected primary monocytes from multiple volunteers, and their cell lysates were analyzed using 2D-DIGE global proteomics. A catalytically inactive GzmA variant was produced to study the enzymatic role in the Mycobacterial Growth Inhibition Assay (MGIA). In parallel, confocal microscopy experiments looked at GzmA internalization. We have evidence that Granzyme A’s intact active site is not necessary to inhibit the intracellular replication of mycobacteria as the S195A variant inhibits mycobacterial infection in MGIA as Wild Type (WT) GzmA. The proteomic approach identified two pathways: ER-stress response and purinergic channel receptor activation are important for GzmA-mediated intracellular mycobacterial inhibition. These results were validated in separate volunteers by western blot. Confocal microscopy experiments show that GzmA is internalized within the infected cell and clears the infection. Modulation of key proteins in the discovered pathways are being targeted to confirm our proteomic results and to test potential novel therapies that emulate GzmA. It will also be investigated if GzmA co-localizes in these cellular compartments. These studies will lead to a better understanding of GzmA biology during infection and to the development of novel host-directed therapeutics for control of Mtb infection.
Combination of dmLT and MPL-A adjuvants synergistically and differentially activates the canonical inflammasome in DCs and macrophages in the absence of pyroptosis
Authors: Bauer David Laurence , Immethun Vicki E , McLachlan James B 
Published: February 2023

Abstract
The initiation of innate immune responses is critical for the formation of a protective adaptive immune response following vaccination. Innate immune cells, such as dendritic cells (DCs) and macrophages, phagocytose and display antigen, produce immune modulators and migrate to lymphoid tissues, leading to the activation of naïve T cells. These professional antigen-presenting cells are activated in response to pathogen or danger associated molecular patterns, a process that is normally not robust with vaccine antigen alone. Adjuvants are able to increase the immunogenicity of vaccine antigens by influencing the activation status of these innate immune cells. Only a few vaccine adjuvants are currently approved for use in the US and, along with the discovery of novel adjuvants, combination adjuvants are likely to have a major impact on future vaccine development. Here, we investigated the ability of both the double mutant Escherichia coli heat labile toxin (dmLT) and the TLR4 agonist monophosphoryl lipid A (MPL-A) to drive innate and adaptative immune responses independently and in combination. We found that the combination adjuvant induces more robust activation of DCs and macrophages than either adjuvant alone, leading to the formation of the NLRP3 inflammasome complex and GSDMD independent secretion of IL-1b in the absence of pyroptosis. Furthermore, RNA sequencing and ingenuity pathway analysis support that the combination adjuvant may allow these DCs to polarize a multi-faceted Th1/2/17 CD4 T cell response. Because macrophages and DCs have distinct roles in the interface between innate and adaptive immunity, understanding the cellular and molecular actions of adjuvants on these cells is critical when developing new vaccines.
Expression and functional characterization of protease-activated receptor 2 (PAR2) and PAR1 on human skin mast cells
Authors: Fukuoka Yoshihiro , Sanchez-Munoz Laura B , Dellinger Anthony L , Schwartz Lawrence B 
Published: February 2023

Abstract
Mast cells play an important role in the pathophysiology of asthma and allergic disease by releasing proinflammatory mediators and cytokines. Activation of protease-activated receptors (PARs) may also play a role in inflammatory response. We investigated the expression of PAR1, 2, 3 and 4 as well as the function of PAR1 and 2 in cultured human skin-derived mast cells (SMCs). Using a combination of flow cytometric analysis, reverse transcriptase-polymerase chain reaction and Western blotting, we demonstrate that human SMCs express PAR1, 2, 3 and 4. Confocal microscopy indicates PAR2 partially colocalize with tryptase within SMCs. Exposure to PAR2 agonist, trypsin and activating peptide (AP) 2-Furoyl-LIGRLO-NH2 induce Ca2+ mobilization in SMCs. The PAR1 APs, SFLLRN-NH2 and TFLLR-NH2 cause degranulation and induce GM-CSF and IL-8 synthesis. Similarly, the PAR2 APs, SLIGKV-NH2, 2-Furoyl-LIGRLO-NH2 also cause degranulation and induce GM-CSF and IL-8 synthesis in SMCs. Both PAR1 APs and PAR2 APs induce rapid phosphorylation of ERK and pertussis toxin (PTX) robustly inhibits phosphorylation of ERK by both PAR1 APs and PAR2 APs. These studies indicate that human SMCs can be activated by both PAR1 and PAR2 which may contribute to MC-mediated inflammatory process.
MicroRNA-155 regulates MAIT cell development, effector subsets differentiation and function
Authors: Liu Tingting , Wang Jie , Subedi Kalpana , Yi Qijun , Mi Qing-Sheng , Zhou Li 
Published: February 2023

Abstract
Mucosal-associated invariant T (MAIT) cells are innate-like T cells that develop in the thymus through sequential maturation stages and differentiate into MAIT1 (T-bet+) and MAIT17 (RORγt+) subsets. MAIT cells have emerged in recent years as crucial players in the effective mucosal immunity and the association of a broad spectrum of disease development. However, detailed molecular mechanisms in regulating MAIT cell development and function remain unclear. Recent studies indicate the involvement of microRNAs in MAIT cell regulation, but few individual miRNAs have been identified, that play the role. To investigate the role of microRNA-155 (miR155) in the development of MAIT cells, we used both gain- and loss-of-function strategies. Using thymus-specific miR-155 knock-in mouse model, it was identified that overexpression of miR-155 significantly reduced numbers and frequencies of MAIT cells in thymus and all peripheral lymphoid and mucosal tissues. Furthermore, MAIT cells with miR-155 overexpression showed interrupted maturation with down regulated PLZF expression and reversed CD8+/CD4−CD8− MAIT subsets distribution. Strikingly, miR-155 overexpression perturbed MAIT17 cell differentiation, and transiently induced miR-155 overexpression inhibited peripheral MAIT cells IL-17 production. Using conventional miR-155 deletion mice model, we found that germline miR-155 deletion did not significantly interrupting MAIT cell development and peripheral tissue MAIT cell differentiation and function. Overall, our results indicate that adequate miR-155 expression is required for MAIT cell development, effector subsets differentiation and function.
Microglial autophagy-mediated clearance of amyloid-beta plaques is dysfunctional in Alzheimer’s Disease mice
Authors: Daily Kylene P , Amer Amal O 
Published: February 2023

Abstract
Alzheimer’s Disease (AD) is a neurodegenerative disorder with no preventative methods or cure. Deposition of aggregated amyloid-beta (Aβ) in the brain is hypothesized to initiate a vicious cascade, leading to inflammation and tissue damage. Microglia are brain-resident immune cells which help maintain homeostasis and clear protein aggregates, including Aβ. However, in AD, microglia produce inflammatory mediators and do not adequately clear Aβ. This project seeks to determine how healthy microglia clear Aβ from the brain and why diseased AD microglia do not. Autophagy is a homeostatic process providing the cell with energy during stress. Autophagy can act as a defense mechanism, by clearing foreign particles and defective proteins. Previous research on AD established that autophagy is dysfunctional in neurons, but autophagy is not well-characterized in AD microglia. Functional autophagy requires expression of multiple autophagy proteins. We hypothesized that AD microglia fail to clear Aβ due to reduced expression of autophagy proteins and that restoring expression of these proteins will enhance autophagy activity. Utilizing microglia from adult AD model mice and various autophagy mice, we demonstrated that AD microglia do not degrade Aβ due to reduced expression of autophagy proteins, which are necessary for Aβ degradation. We identified an upregulated cluster of microRNA in microglia from mouse and human AD brain tissue samples. Inhibiting these microRNA improves expression of specific autophagy proteins and restores Aβ degradation in AD mouse microglia. Our findings indicate that autophagy is dysfunctional in AD microglia and establish a mechanism behind dysfunctional autophagy which may be therapeutically targeted.