bluuebunny/crossref_metadata_2025_split huggingface dataset:
Therapeutic efficacy of statins and AICAR in mitochondrial dysfunction in EAE
Authors: Singh Inderjit , Samuvel Devadoss J. , Singh Avtar K. , Won Jeseong 
Published: January 2023

Abstract
Studies from various laboratories have provided evidence of mitochondrial dysfunction as a cause of the CNS pathology in multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). Recently, we described that RhoA-ROCK-PPAR mediated activity of statins as well as activation of AMP activated protein kinase (AMPK) protect oligodendrocytes against inflammatory insults in EAE. EAE disease decreased the activation of AMPK whereas combination of lovastatin and AICAR, a specific activator of AMPK, improved the efficacy against clinical disease of EAE. As expected, mitochondrial activities of electron transport chain complexes and thus levels of ATP were decreased in the CNS of EAE as compared to control animals whereas treatment with combination of lovastatin and AICAR corrected the EAE induced mitochondrial dysfunctions while individual these drugs were partially effective. The loss of mitochondrial functions in EAE paralleled the decrease in decreased biogenesis of mitochondria as decreased in mitochondrial biogenesis proteins (PGC1α, SIRT1, and NRF1). Treatment of EAE animals with combination of AICAR and lovastatin corrected the expression of these proteins as well as increased the expression of PPARα/γ. These corrections of mitochondrial dysfunctions paralleled the levels of myelin proteins. These finding suggest that simultaneous targeting of RhoA-ROCK-PPAR signaling mechanisms by lovastatin and AMPK/PGC1α signaling mechanisms with AMPK activator AICAR provides greater neuroprotection, neurorepair, and hence improved efficacy. These findings highly relevant to improve the efficacy of statins observed in a recent study of patient with progressive MS (Chataway et al., Lancet 383:2213, 2014).
Manganese-enhanced magnetic resonance imaging detects VIPR2-mediated immune transformation in a model of Parkinson’s disease
Authors: Olson Katherine E , Schutt Charles , Bade Aditya , Dong Jingdong , Boska Michael , Shandler Scott , Mosley R. Lee , Gendelman Howard E , Liu Yutong 
Published: January 2023

Abstract
Progressive nigrostriatal degeneration and neuroinflammation underlie the pathobiology of Parkinson’s disease (PD). Biomarkers to detect such pathogenic events remain in development. We report that manganese-enhanced magnetic resonance imaging (MEMRI) can be used to assess inflammatory activities related to deficits in neural connectivity in an experimental PD model. Vasoactive intestinal peptide (VIP), an endogenous neuropeptide that binds to VIPR1 and VIPR2 inducing a phenotypic shift of neurotoxic effector memory to neuroprotective regulatory T cells was modified (referred to as LBT-3627) to improve its half-life and limit proteolytic cleavage. LBT-3627 was used to investigate its ability to elicit neuroprotection in mice treated with the nigrostriatal toxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Treatment with LBT-3627 lead to neuronal sparing when compared to MPTP-treated mice. Seven days post MPTP intoxication, mice were scanned using MEMRI, and images were registered to an MEMRI-based brain atlas for comparison. Manganese-induced MRI signal enhancement was calculated. MEMRI detected MPTP-induced inflammation as well as a corresponding reduction in signal with low and high doses of LBT-3627. MPTP-treated mice showed a significantly higher signal enhancement within the hippocampus (p = 0.04), with a trend towards significance in both the substantia nigra and striatum when compared to saline controls. The higher signal enhancement was reversed by LBT-3627 treatment. These findings indicate the ability of MEMRI to detect MPTP-induced brain pathology and its reversal following LBT-3627 treatment, supporting MEMRI as a method to assess changes in nigrostriatal-induced neuroinflammation.
TLR and CD40 signalings synergize to induce negative regulators that inhibit B cell class-switching and plasma cell differentiation: relevance to antibody responses
Authors: Wang Rui , Lou Zheng , Casali Paolo , Xu Zhenming 
Published: January 2023

Abstract
During host immune responses to microbial pathogens and allergens, a B cell is exposed to T-independent (e.g. TLR ligands) and T-dependent (e.g., CD40 ligand, CD154) stimuli and integrates different signals for effective antibody responses. We have previously shown that engagement of TLRs (e.g., TLR4 by LPS and TLR9 by CpG) or CD40 induces Ig class switch DNA recombination (CSR) and B cell differentiation into plasma cells, albeit to different degrees. Here, we showed that combined stimulation of B cells through TLR and CD40 (by LPS plus CD154 or CpG plus CD154) abrogated CSR to IgG1, IgG2b, IgG3, IgA and IgE as well as plasma cell differentiation. Combined TLR/CD40 stimulation, however, did not significantly affect IgH germline IH-CH transcription or AID expression, both of which are critical for CSR, or expression of Blimp-1, a master transcription factor in plasma cell differentiation. Rather, LPS and CD154 synergized to upregulate (up to 3000-fold) expression of genes encoding EBI3 and p28, the two subunits of the cytokine IL-27. An anti–p28 blocking antibody could partially restore CSR and plasma cell differentiation in TLR/CD40 co-stimulated B cells. This together with CSR rescue by transduction of replication-defective lentiviral particles, which could inhibit IL-27 functions, indicates a negatively regulating role of IL-27 in B cell differentiation. It also suggests that engagement of TLRs and CD40 by microbial components and T helper cells may shape the antibody response by modulating B cell-intrinsic functions.
Supported by NIH grants AI 079705 and AI 105813 grants to P.C.
Asynchronous digital activation of the germline Iγ1 and Iɛ promoters in single cells reveals that temporal control determines the fate of switching to IgE in B cells
Authors: Wu Yee Ling , Stubbington Mike , Teichmann Sarah , Rada Cristina 
Published: January 2023

Abstract
Immunoglobulin IgE is a key effector in the adaptive immune response to parasitic infestations and a potent mediator of allergy and asthma. Switching to IgE is linked to the activation of the germline promoter (Iɛ) that drives transcription through the region (Sɛ) immediately upstream of the ɛ constant region and the recruitment of Activation Induced Deaminase (AID) to induce double strand breaks. The molecular basis of AID recruitment to Sɛ and thus the specificity of switching to IgE remains elusive given that the same stimuli that activate Iɛ also initiate transcription from the Iγ1 promoter upstream of the γ1 constant region.
We have used knock-in fluorescent reporter mice for the endogenous Iγ1 and Iɛ promoters to monitor the kinetics of in vitro I-promoter activation by cytokines and isotype switching. While the activation of I-promoters is biallelic, a significant number of activated cells (2/3) displays asynchronous activation, with activation of a single Iγ1 allele followed by activation of the second allele over a period of 12–24 hrs in vitro. Intriguingly, early activation of the Iγ1 promoter is unaffected by the rearrangement status of the cis VDJ region.
Single-cell transcriptomic and qPCR analyses on activated B cells reveal enormous heterogeneity in I-promoter activation within individual cells, identifying a distinct early population of cells in which only the Iɛ promoter becomes active, which is correlated with enhanced switching to IgE. Heterogeneity of allele activation in single cells is emerging as a general property of transcriptional programming. In B cells, the intrinsic temporal heterogeneity of I-promoter activation in individual cells is crucial for the control of AID targeting and the levels of IgE switching.
Control of B cell activation and migration by PI 3-kinase: role of inositol polyphosphate 4-phosphatases
Authors: Marshall Aaron J , Hou Sen , Wu Xun , Li Hongzhao 
Published: January 2023

Abstract
Balanced activation of the PI3K pathway by antigen and other stimuli is required for effective humoral immunity while avoiding autoimmunity and lymphoproliferative disease. The critical regulatory phosphatase SHIP dephosphorylates the PIP3 generated by PI3K to produce a distinct molecule PI(3,4)P2. A number of key signaling molecules bind to PIP3, including the protein kinase Btk; however much less is known about the function of PI(3,4)P2. The inositol phosphatase INPP4A can specifically hydrolyze PI(3,4)P2; however the functions of this phosphatase in B cell activation are unknown. We hypothesized that INPP4A can provide a tool to deplete PI(3,4)P2 and determine its range of functions. Human B lymphocytes over-expressing active or phosphatase-dead INPP4A were generated, and the cells expressing active enzyme were found to generate lower levels of PI(3,4)P2 upon chemokine or BCR stimulation. Chemokine-induced migration responses were inhibited by active, but not inactive INPP4A. An assessment of BCR signaling was carried out that compared BCR-stimulated cells expressing active versus inactive INPP4A in a screen encompassing over 800 protein phosphorylation sites. The results indicate that active INPP4A can suppress phosphorylation of Akt and known Akt targets. We are currently assessing whether PDK1, a major kinase upstream of Akt which also directly binds PI(3,4)P2, is regulated by INPP4A. Interestingly, other kinases known to interact with PDK1 (PKCdelta, RSK) or to be directly phosphorylated by PDK1 (PKCgamma) showed decreased phosphorylation in cells expressing active INPP4A. We are currently testing the hypothesis that PDK1/Akt and Btk/PLCγ2 signal osomes are differentially regulated by INPP4A.
Toll-like receptor 9 signaling antagonizes the B cell receptor-dependent ability of B cells to process and present antigen to helper T cells.
Authors: Akkaya Munir , Kim Ann S , Miozzo Pietro , Sohn Hae-Won , Akkaya Billur , Pena Mirna , Pierce Susan K 
Published: January 2023

Abstract
B cells express both an adaptive antigen-specific B cell receptor (BCR) and innate Toll-like receptors (TLRs) allowing the functional outcome of the B cell’s engagement with antigen to be modulated in response to pathogen-derived TLR ligands. In T cell-dependent responses, the BCR both signals for B cell proliferation and differentiation and also internalizes bound antigen for processing and presentation to helper T cells. Here we show that although the presence of the TLR9 ligand, CpG, does not affect early BCR mediated events following antigen binding including phosphorylation of key components of the signaling cascade and the formation of BCR clusters on cell surface, CpG alters the outcome of BCR signaling, resulting in unique transcriptional profiles, enhanced proliferation and differentiation to antibody secreting cells. In contrast, CpG dramatically limits the ability of B cells to process and present antigen to helper T cells. In the presence of CpG, BCR-induced upregulation of CD86 and MHC class II expressions are antagonized and antigen internalized by the BCR is not properly trafficked to antigen processing compartments resulting in reduced numbers of peptide-MHC class II complexes on the B cell surface. Indeed, CpG treated antigen-specific B cells show a reduced ability to maintain contact with antigen-specific T cells and to activate antigen-specific helper T cell proliferation in vitro. These results suggest that TLR9 activation in T cell dependent responses would drive B cell toward proliferation and antibody secretion and away from events that are highly dependent on the ability of B cells to present antigen to helper T cells, such as entry into germinal centers and affinity selection.
Immune modulation by butyrophilin 1A1 (BTN1A1)
Authors: Wang Jinghua , Manick Brian , Wu Guoping , Kalabokis Vassili 
Published: January 2023

Abstract
The butyrophilins are B7-like T cell co-regulatory molecules. These molecules are of increasing interest in cancer immunotherapy as they may represent a novel subset of immune check-point regulators. In this study, we expressed the recombinant human butyrophilin 1A1 extracellular domain with a C terminal 6-His tag (BTN1A1) in NSO mouse myeloma cell line, and investigated the biological functions of the purified recombinant BTN1A1 protein in vitro. Human T cells were treated with plate-bound anti-CD3 and BTN1A1. The levels of multiple cytokines were measured in cell culture supernatants using R&D Systems Proteome Profiler™ Human Cytokine Array and confirmed by measuring individual cytokines using cytokine-specific Quantikine® ELISA kits. BTN1A1 significantly decreased T-cell derived cytokines such as IL-2, IL-21, IL-25, CD40L, and C5a, but not IFN-gamma. BTN1A1 also markedly inhibited anti-CD3-induced human T cell proliferation in a dose dependent manner. Furthermore, BTN1A1 showed binding to anti-CD3 activated, but not rest T cells. Moreover, recombinant human BTN1A1 inhibited p38 MAPK, CREB, and TOR Phosphorylation, suggesting BTN1A1 modulates TCR signaling through the p38 MAP kinase, CREB, and TOR pathways. Taken together, our data suggests that BTN1A1 acts as a co-inhibitory molecule to modulate T cells through an unknown receptor on the surface of T cells. BTN1A1 may be a potential target of immune check-point molecules for therapeutic purposes.
Characterization of a new lymphoid organs-specific endothelial cells subset with high expression of VISTA negative co-stimulator protein
Authors: Salehi Maryam , Radichev Ilain , Savinov Alexei 
Published: January 2023

Abstract
Peripheral tolerance is regulated through a network of co-stimulatory and co-inhibitory ligands and receptors. Targeting these molecules or their pathways has been promising in clinic in both oncoimmunology and autoimmunity. V-set domain Ig Suppressor of T cell Activation (VISTA) is a recently identified co-inhibitory member of the B7 family. VISTA suppresses T cell immune responses; interference with its function results in susceptibility to autoimmunity. Investigating expression pattern of VISTA in various cell types and tissues in mice, we identified a sub-population of endothelial cells present only within secondary lymphoid organs, which express high levels of VISTA; a finding not previously reported. Characterization of this subset revealed that VISTA-expressing endothelial cells (VEECs) are predominantly of lymphatic endothelial cells (LECs) origin, however, only less than half of LECs in lymphoid organs are VEECs. As LECs have been shown to induce tolerance via co-inhibitory molecules such as PD-L1, we studied the role of VEECs in regard to immune tolerance. FACS-sorted VEECs cells from secondary lymphoid organs of NOD and DBA mice were evaluated for the expression levels of various peripheral tissue antigens associated with immune tolerance. Our results show that in NOD VEECs the AIRE transcription factor, as well as some diabetic autoimmunity-associated peripheral tissue antigens of pancreatic origin (insulin-2 and Rrad) are significantly up-regulated. To further investigate the regulatory role of VEECs, we applied co-culture assays and evaluated the effects of VISTA expression by endothelial cells on T cell function.
Defining the phenotype and transcriptome of IgG3+ B cells
Authors: Sadanand Saheli , Yu Wen-Han , Alter Galit 
Published: January 2023

Abstract
Recent studies suggest that highly polyfunctional non-neutralizing HIV-specific IgG3 antibodies are associated with protection from HIV infection in the RV144 HIV vaccine trial. While it is well established that IgG3 Abs can drive a wide range of Fc-mediated innate effector functions, it remains unclear whether intrinsic factors are important for selection and maintenance of IgG3-secreting B cells and what extrinsic signals preferentially induce IgG3 class-switch. We hypothesized that (1) IgG3+ B cells will generally express enhanced levels of activation-associated surface proteins and markers important for interactions with T cells as compared to IgG1+ B cells and (2) IgG3+ B cells will have a unique transcriptome that helps prevent downstream class switch recombination. Our initial flow cytometry and RNA-sequencing (RNA-seq) analyses have focused on comparing IgG3+ memory B cells to IgM+ and IgG1+ memory B cells in HIV negative donors. We have found that IgG3+ B cells have higher expression of CD80 and CD95, but lower expression of CD40, than IgG1+ B cells. Preliminary bulk RNA-seq analyses similarly suggest that IgG3+ B cells have a unique transcriptome, with distinct expression of genes involved in transcriptional regulation, nucleosome assembly and chromatin organization. Interestingly, IgM+ and IgG1+ memory B cells share an overlapping transcriptional regulation cluster that may exist to prevent class-switch to IgG3 (from IgM) and induce class-switch to IgG1 (from IgG3). We are currently investigating the heterogeneity of resting and stimulated IgG3+ memory B cells at the single-cell level to better understand why some, but not all, IgG3+ memory B cells retain expression of IgG3.
HSP90B1 Regulates TLR-dependent Monocyte Signaling and its Common Variants are Associated with BCG-specific T-cell Responses and Protection from Pediatric TB Disease
Authors: Graustein Andrew , Misch Elizabeth A , Musvosvi Munyaradzi , Shey Muki , Shah Javeed , Wells Rick , Hanekom Willem , Hatherill Mark , Scriba Thomas , Hawn Thomas 
Published: January 2023

Abstract

Introduction
HSP90B1 is a chaperone for multiple Toll-like receptors (TLRs), regulates innate immune signaling in murine myeloid cells and is expressed in T-cells. We hypothesized human HSP90B1 regulates monocyte responses to Mycobacterium tuberculosis (Mtb) and that its variants are associated with T-cell responses to the BCG vaccine and susceptibility to TB disease.

Methods & Results
We screened 17 haplotype-tagging HSP90B1 gene region SNPs for association with BCG-induced immune responses in South African infants vaccinated with BCG at birth. We stimulated peripheral blood drawn at 10 weeks of age with BCG in a whole blood cytokine assay. Three HSP90B1 SNPs (rs10507172, 10507173, and 2164747) were associated with increased IL-2 secretion (P&lt;0.05, generalized linear model). SNPrs10507173 was associated with increased IL-2 production in the CD4+ T-cell subset (p=0.028, ANOVA) based on intracellular cytokine staining and flow cytometry. The same three alleles were associated with protection from pediatric TB disease in a gene association study of 226 children who contracted tuberculosis compared with 626 controls who remained healthy (dominant model, unadjusted p&lt;0.05, OR 0.58–0.72; after adjusting for ethnicity, p&lt;0.05 for 2/3 SNPs). We used CRISPR/Cas9 gene editing to reduce HSP90B1 expression in U937 monocyte cells. Relative to controls, HSP90B1-deficient cells secreted less TNF when exposed to TLR1/2/6 (p=0.02) and TLR4 (p=0.01) ligands and Mtb whole cell lysate (p=0.001).

Conclusion
These data suggest HSP90B1 deficiency is associated with an impaired monocyte response to Mtb ligands. In infants, HSP90B1 variants are associated with increased BCG-specific IL-2 responses T-cells and protection from TB disease.
Aspirin Triggered Lipoxin A4 regulates NF-κB and induces antimicrobial peptides expression in airway epithelial cells to limit mucosal inflammation to enhance host response in pneumonia
Authors: Sham Ho Pan , Abdulnour Raja-Elie E , Krishnamoorthy Nandini , Douda David N , Colby Jennifer K , Levy Bruce D 
Published: January 2023

Abstract
Bacteria induced pneumonia remains a major global threat to health. Even with effective antibiotic treatment, the host response can be exuberant, resulting in tissue damage and morbidity. Resolution of inflammation is an active process governed in part by specialized pro-resolving mediators (SPMs). We hypothesized that the SPM, aspirin triggered lipoxin A4 (ATL) engaged airway mucosal epithelial cells to enhance resolution. In a murine model of bacterial aspiration pneumonia for E. coli, ATL (100 ng, iv) decreased the lung bacterial burden in an additive manner with ciprofloxacin (0.2 mg/kg, ip). Further, ATL also significantly limited neutrophils recruitment and decreased pro-inflammatory cytokine (IL-1α) and chemokine production (KC). These ATL mediated protective responses were mediated in part via regulation of NF-kB activation. In response to LPS and E. coli in vitro, ATL selectively influenced airway epithelial cells expression of NF-kB regulators A20, SIGIRR and ST2. In an in vitro bacteria killing assay by airway epithelial cells, ATL directly limited bacterial growth by increasing select antimicrobial peptides expression (LL37 and BPI). mCRAMP deficient mice (human homolog of LL37) failed to limit lung bacterial growth in E. coli pneumonia and ATL treatment was significantly less efficient in promoting host defense in these mice. In conclusion, the SPM ATL can limit pathogen mediated mucosal inflammation and enhance host defense in pneumonia via the selective regulation of NF-κB and antimicrobial peptides in airway epithelial cells. These findings with ATL provide evidence for SPM targeted treatment of the host response to pneumonia that has the potential for therapeutic benefit in combination with antibiotics.
The impact of bacterial superinfection during influenza infection of allergic hosts
Authors: Samarasinghe Amali , LeMessurier Kim , Iverson Amy , Rosch Jason , McCullers Jonathan 
Published: January 2023

Abstract
Allergic asthma can be complicated by respiratory pathogens. However, asthmatics suffered less severe morbidity and mortality compared to non-asthmatics during the 2009 influenza A virus (IAV) pandemic. Using a novel mouse model, we determined that IAV infection during allergic airways inflammation protects the host from influenza. Pathogen-pathogen interactions between IAV and Streptococcus pneumoniae (Spn) can lead to fatal outcomes in a healthy host. We hypothesized that allergic hosts will have less morbidity and mortality associated with Spn superinfection after influenza. Infectious IAV was undetectable in the lungs at 72 h after Spn. Similar to animals infected with Spn alone, asthma+Spn mice cleared the bacteria completely by 72 h. Bacterial burden increased in the bronchoalveolar lavage fluid (BALF), lungs, and blood of mice in the IAV+Spn co-infection group. However, bacterial load in animals subjected to the fungal asthma model prior to viral and bacterial infections were significantly reduced in the BALF and absent in the lungs and blood. More leukocytes were found in the BALF of mice infected with IAV. The number of macrophages was similar between all groups, while those in the asthma+IAV, asthma+Spn, asthma+IAV+Spn groups had prominent amounts of eosinophils and CD4+ T cells (30% each) in the airways which was significantly higher than the asthma-only, IAV-only, Spn-only, and IAV+Spn groups. Neutrophils were most abundant in the IAV+Spn controls while the greatest number of CD8+ T cells was present in the IAV-only controls. The lung microbiome was significantly different in asthma+IAV+Spn. Remarkably, the asthma+IAV+Spn group did not have severe morbidity or mortality typically associated with IAV+Spn coinfection.