bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rictor positively regulates B cell receptor signaling by modulating actin reorganization via ezrin
Authors: Liu Chaohong , Huang Lu , Zhang Yongjie , Zhao Xiaodong 
Published: January 2023

Abstract
As the central hub of the metabolism machinary, mTORC2 has been well studied in lymphocytes. As an obligatory component of mTORC2, the role of Rictor in T cells is well established. However, the role of Rictor in B cells still remains elusive. It has been reported that Rictor is involved in B cell development, especially the peripheral development. But the role of Rictor on BCR signaling as well as the underlying cellular and molecular mechanism is still unknown. This study used B cell specfic Rictor knockout mice (cd19Cre) to investigate how Rictor regulates BCR signaling. We found that for the key positive and negative BCR signaling molecules, pBtk is reduced and pSHIP is enhanced in Rictor KO B cells. This indicates Rictor positively regulates the early events of B cell receptor (BCR) signaling. We found that the cellular F-actin is drastically increased in Rictor KO B cells after BCR stimulation through dysregulating the dephosphorylation of ezrin. The high actin-ezrin intensity area restricts the lateral movement of BCRs upon stimulation, consequently reducing the BCR clustering and BCR signaling. The reduction in the initiation of BCR signaling caused by actin alteration leads to decreases in the humoral immune response in Rictor KO mice. The inhibition of actin polymerization with Latrunculin in Rictor KO B cells rescues the defects of BCR signaling and B cell differentiation. Overall, our study provides a new pathway linking cell metablism to BCR activation, where Rictor regulates BCR signaling via actin reorganization.
Itk independent development of natural Th17 cells and IL17A-driven hypersensitivity pneumonitis
Authors: Elmore Jessica , Huang Weishan , Carter Chavez , Koylass Nicholas , Bennett Amelia , August Avery 
Published: January 2023

Abstract
The tyrosine kinase, Itk, plays a critical role in T cell receptor signaling, and we have previously shown that Itk is required for the development of Th17 cells and their production of the pro-inflammatory cytokine IL17A. IL17A has been implicated in numerous pulmonary inflammatory diseases. We have previously shown that Itk−/− mice do not develop allergic airway disease coupled with protected reduced production of IL17A in the lungs in a model of allergic asthma. Farmer’s lung is a subset of hypersensitivity pneumonitis caused by repeated exposure to the bacteria Saccharopolyspora rectivirgula (SR). This IL17A-driven disease is characterized by an influx of neutrophils and T lymphocytes. Here we show that a population of natural Th17 cell develop in the absence of Itk. Furthermore, exposure to SR drives robust IL17A response produced by CD4+ T cells even in the absence of Itk, accompanied by the development of pulmonary inflammation. These studies suggest that TCR signaling through Itk differentially regulates the development of conventional and natural Th17 cells. Understanding how Itk modulates the Th17 cytokine responses will allow us to better understand the precise role of Itk in the production of IL17A in airway inflammation.
Targeting class switch recombination plays a potential role in retarding systemic lupus erythematosus (SLE)
Authors: Zhu Jing , Wagner Alayna , Kasmark Leah , Woodward Emily , Sproule Tom , Roopenian Derry C , Leeth Caroline McPhee 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease mediated mainly by autoantibodies reactive to nuclear acids and proteins. These autoantibodies are mostly of IgG isotype, indicating that class-switch recombination (CSR) may play an important role in the development of SLE. CSR is initiated by the enzyme, activation-induced cytidine deaminase (AID) which induces double strand DNA breaks then repaired by homologous recombination using the RAD51 complex. We hypothesize that therapeutic targeting of CSR will retard the development of SLE by inducing cell death in AID-expressed B cells and reducing pathogenic IgG antibodies. The BXSB/MpJ mouse is a well characterized model of human SLE, exhibiting a robust germinal center response and copious production of immunoglobulins. 4,4’-diisothiocyanatostilbene-2-2’-disulfonic acid (DIDS) is a molecule which impairs homologous recombination by inhibiting the binding of RAD51 complex to DNA. In this study, male BXSB/MpJ mice were injected with DIDS or control. Treated mice showed a reduction in the germinal center B cell population and a corresponding decrease in follicular T helper cells. T1 B cell and follicular B cell populations were significantly decreased in DIDS treated mice but no changes were found in marginal zone B cells. Sera were analyzed for the immunoglobulin levels and antinuclear antibodies. These findings demonstrate that targeting CSR by DIDS reduces potentially pathogenic B lymphocyte populations and may be a viable novel therapeutic option in the treatment of SLE.
Autoimmune responses are dependent on the MAVS innate pathway due to its natural co-stimulating effect on B cell activation and maturation
Authors: Sun Wenxiang , Qi Chen-Feng , Scott Bethany , Bolland Silvia 
Published: January 2023

Abstract
There is evidence that autoimmune disease might be induced by deficient regulation of innate immune pathways that normally induce type I interferon in response to pathogen-derived nucleic acids. To investigate a potential role for nucleic acid sensing pathways with cytosolic localization and more ubiquitous expression, we tested the requirement for MAVS (dsRNA-sensing) or STING (dsDNA-sensing) in the lupus autoimmune disease developed by FcγRIIB-KO (R2−/−) mice. These experiments revealed that STING was mildly protective, while MAVS was an essential factor for autoimmune disease in R2−/− mice. We found that MAVS deficiency, but not type I IFN receptor deficiency, completely eliminated autoantibodies and autoimmune pathology in R2−/− mice. MAVS deficiency also eliminated spontaneous activation of B and T cells, germinal center formation and follicular helper T cell development. Using mixed bone marrow reconstitution experiments, we determined that MAVS was required for autoreactivity in a B cell intrinsic manner, with a profound effect on B cell survival in the germinal center. MAVS deficiency did not alter BCR-proximal signaling, but resulted in diminished long term proliferative capacity of B cells, with lower cyclin D2 and Bcl-xL expression 24 hours after anti-IgM stimulation. MAVS deficiency also resulted in B cell developmental defects, fewer total B cells but a skewing toward marginal zone and against the follicular B cell program. Overall MAVS seems to provide its costimulatory effect independent of type I IFN, by enhancing the activation of NFκB in situations where T cell help or other modes of costimulation are not present and most importantly, by providing survival stimulus for autoreactive B cells.
Comprehensive proteomic profiling of Sjogren’s syndrome revealed dysregulation of interferon and other immunologic pathways
Authors: Wang Jingya , Wang Shu , Rahman Saifur , Kumar Varsha , Karnell Jodi L. , Voynova Elisaveta Ninova , Sanjuan Miguel A , Kolbeck Roland , Alevizos Ilias , Ettinger Rachel 
Published: January 2023

Abstract
Sjögren’s syndrome (SS) is the second most common rheumatic autoimmune disease with a broad spectrum of clinical and serological manifestations and no approved biologics. To better understand the pathogenesis of SS and explore potential soluble biomarkers and therapeutic targets, sera from 109 SS and 50 healthy controls (HC) were examined. Comprehensive profiling was performed using autoantigen and proteomic arrays (UTSW, 95 autoAb’s; RBM’s, 165 proteins; and SOMAscan, 1129 protein). Data showed that autoAb’s against U1sn-RNP-BB’, U1sn-RNP-C and Ribo phaspho protein P1 significantly increased in SS compared to HC (Fold change &gt;1.5 and FDR &lt;0.05) that positively correlate with FOCUS disease score (FDR&lt;0.05). RBM and SOMAscan was highly consistent and detected 123 unique proteins up-or down-regulated in SS compared to HC. Strikingly, 71 were interferon-regulated proteins, with IP10 correlating best with FOCUS score (r=0.65). Pathway and network analysis also confirmed strong activation of type I interferon signaling. Type I interferon signature genes were also observed in salivary gland of the NOD.H-2h4 mice, a model that closely resembles human SS. Additional proteins dysregulated in SS patent sera include the TNF signaling, antimicrobial proteins A2M and B2M, and multiple SLAMF family members, which may shed light on disease mechanisms and novel therapeutic strategies.
Human MEMs are a distinct high IL-6 producing macrophage subset with anti-inflammatory and radioprotective properties in vivo
Authors: Capitini Christian M. , Bouchlaka Myriam N , Moffitt Andrea , Kim Jaehyup , Kink John A , Bloom Debra D , Love Cassandra , Dave Sandeep , Hematti Peiman 
Published: January 2023

Abstract
Macrophages (MØs) can polarize into subsets of “pro-inflammatory” (M1) MØs or “anti-inflammatory” (M2) MØs. Previously we described a unique population of MØs, termed human mesenchymal stem cell (MSC)-educated MØs (MEMs). In this study, we hypothesize that MEMs have potential to attenuate inflammation and promote tissue repair in vivo based on their immunophenotype and secretome. MEMs were generated by isolating monocytes from human peripheral blood and culturing them into MØs, followed by a co-culture with bone marrow-derived MSCs. RNA-Seq analysis showed genes associated with epithelial-mesenchymal transition, collagen formation, tissue development, and cell-cell adhesion were positively correlated with MEMs, while genes associated with oxidative phosphorylation were negatively correlated as compared to M1 MØs. We found increased expression of 8 genes implicated in wound healing, as well as genes representing remodeling enzymes, growth factors and the cytoskeleton. RT-PCR verified MEMs have increased CD206 (p&lt;0.0001), IL-1β (p&lt;0.001), TGFβ (p&lt;0.001) and arginase-1 (p&lt; 0.05), as well as increased IL-6 (p&lt;0.0001) and CD73 (p&lt;0.001) expression. MEMs produced over 4x greater amounts of IL-6 as compared to M2 MØs by ELISA (p&lt;0.001), which could be enhanced with LPS stimulation (p&lt;0.0001) or attenuated by inhibition of COX-2 (p&lt;0.05), arginase-1 (p&lt;0.01) or JAK1/JAK2 (p&lt;0.05). In vivo, MEMs improved survival against lethal GVHD (p&lt;0.01), in part by reducing T cell proliferation (p&lt;0.001). MEMs also enhanced survival of mice from lethal radiation injury (p&lt;0.01), in part by inducing host fibroblast proliferation (p&lt;0.01). MEMs are a unique MØ subset with therapeutic potential for inflammatory disorders.
Molecular-based diagnosis of Multiple Sclerosis and its progressive stage
Authors: Kosa Peter , Barbour Christopher , Komori Mika , Tanigawa Makoto , Wu Tianxia , Johnson Kory , Masvekar Ruturaj , Fossati Valentina , Douvaras Panagiotis , Herbst Ronald , Wang Yue , Tan Keith , Greenwood Mark , Bielekova Bibiana 
Published: January 2023

Abstract
Biomarkers aid diagnosis, allow inexpensive screening of therapies and guide selection of rational patient-specific polypharmacy regimens in most internal medicine disciplines. In contrast, there are no clinically-validated measurements of the physiological status, or dysfunction(s) of cellular components of the central nervous system (CNS). Accordingly, patients with chronic neurological diseases are generally treated with a single disease-modifying therapy without understanding patient-specific drivers of disability.
Therefore, using multiple sclerosis (MS) as an example of a complex polygenic neurological disease, we sought to determine if cerebrospinal fluid (CSF) biomarkers are intra-individually stable, cell type-, disease- and/or process-specific and responsive to therapeutic intervention.
Using DNA-aptamer-based multiplex assay, in-vitro- and statistical models, we developed in a training cohort (n=225) and validated in an independent-cohort (n=85) two biomarker-based classifiers that differentiate: 1. MS from CNS diseases that mimic MS clinically, pathophysiologically and on imaging, and 2. relapsing-remitting from progressive MS with ~90% accuracy. Treatment-induced changes in biomarkers greatly exceeded intra-individual-and technical variabilities of the assay.
The observation that CNS biological processes reflected by combinatorial CSF biomarkers are robust, stable and disease- or even disease-stage specific opens opportunities for broad utilization of CSF biomarkers in drug development and precision medicine for CNS disorders.
Detection of fg/mL Levels of IL-2, IL-4, IL-6, IL-10, and IL-17A in Serum, Plasma, and Supernatant of Established Cell Models
Authors: Georlette Daphne , Wheeler Sarah E , Ranganathan Sripriya , Wang Su , Nikolenko Galina N , Shukla Animesh , Miller Thomas C , Aghvanyan Anahit , Stengelin Martin , Mathew Anu , Kuklis Catlin , Sharma Deepak , Bess Matthew , Davydov Ilia V , Wohlstadter Jacob N 
Published: January 2023

Abstract
Most immunoassays quantify cytokines with pg/mL sensitivity, which is insufficient to detect those present at low levels. MSD’s next-generation S-PLEXSM assay format was developed to quantify cytokines with fg/mL sensitivity. Here we describe measuring IL-2, IL-4, IL-6, IL-10, and IL-17A levels in normal sera and the supernatants of model cell lines using S-PLEX assays.
The assays demonstrated a dynamic range of approximately four orders of magnitude. Standard intra-plate coefficients of variation (CVs) ranged from 3%-15% and inter-plate CVs ranged from 8%–18%. The lower limit of detection (LLOD) was &lt;1 fg/mL for all assays except IL-17A (11 fg/mL). IL-2, IL-6, IL-10, and IL-17A were detectable in 100% of normal sera samples (n=36–75) and IL-4 was detectable in 40% of normal sera samples (n=75). The average concentrations were &lt;1 pg/mL for normal samples.
To confirm the sensitivity of these assays and their ability to detect native analytes, we characterized a panel of 23 cell lines that are models for cytokine secretion. As an example, the MOLT-4 cell line derived from acute lymphoblastic leukemia natively expressed detectable levels of IL-2 (1,031 fg/mL), IL-4 (221 fg/mL), IL-10 (60 fg/mL), and IL-17A (30 fg/mL). The expression profiles of these cell lines confirmed the sensitivity of the S-PLEX assays.
S-PLEX cytokine assays provide a 100 to 1,000-fold lower LOD than standard ELISAs, allowing the determination of baseline cytokine levels in many sample types.
Molecular chaperone is required for gut tolerogenic dendritic cell development and function
Authors: Iwanowycz Stephen , Yang Yi , Li Zihai , Liu Bei 
Published: January 2023

Abstract
The intestinal mucosa is continuously inundated with foreign antigens. Therefore, immune cells need to be tightly regulated to maintain tolerance against harmless dietary antigens and microbiota while responding to pathogens. Intestinal dendritic cells (DCs) play an integral role in regulating immunity and tolerance because of their unique position at the interface of innate and adaptive immunity. Multiple populations of intestinal DCs have been reported based on the expression of CD103, CD11b, and CX3CR1. However, the mechanism by which different DC subsets regulate intestinal homeostasis is still unclear. Heat shock protein gp96 is an essential chaperone for most TLRs, integrins, and other proteins important for DC functions. However, the role of DC-intrinsic gp96 in regulating gut tolerance has not been studied. By a genetic strategy, we discovered that selective deletion of gp96 from CD11c+ cells in mice results in alteration of DC and T cell subsets in the gut as well as the loss of antigen-specific regulatory T cell (Treg) induction. Strikingly DC-specific gp96 knockout mice develop spontaneous colitis by 24 weeks of age. Furthermore, differentiation of murine bone marrow-derived DCs revealed that gp96 depletion selectively decreased cell maturation in response to FLT3L compared to CSF2. Also, gp96 depletion reduced DC migration and cross-presentation. Taken together, our results demonstrated that gp96 depletion from CD11c+ cells selectively inhibits tolerogenic DCs in the gut leading to decreased Treg cells, which indicates that DC-intrinsic gp96 plays essential roles in maintaining gut tolerance.
The intestinal lamina propria is a unique niche for regulatory T cells
Authors: Morales Elisa Cruz , Laufer Terri M. 
Published: January 2023

Abstract
Foxp3+ regulatory T cells (Tregs) are critical to mantain intestinal homeostasis. Like all CD4+ T cells, Tregs require TCR-MHCII interactions for their development in thymus and differentiation in periphery. Past studies suggest that the TCR repertoire of intestinal lamina propria (siLP) Tregs is skewed towards food and commensal bacteria antigens. However, by using a transgenic mouse model that lacks peripheral MHCII expression and peripheral Tregs induction (K14 mice), we have demonstrated that thymically generated Tregs could enter to the siLP of weanlings and proliferate independently of MHCII to fill and maintain the compartment.
Newly-generated Tregs egress from thymus as central Tregs and acquire an effector phenotype after TCR signals in periphery. However, despite the lack of MHCII in the siLP, K14 Tregs are activated and phenotypically similar to effector Tregs in WT siLP. Interestingly, transcriptional profiling reveals that many components of a TCR-dependent expression module are present in both K14 and WT siLP Tregs. Furthermore, differently to splenic Tregs, K14 siLP Tregs highly express Nur77 and IRF4, often regulated through TCR signaling.
Tregs from K14 siLP localize mainly at isolated lymphoid follicles, where they are in proximity to B cells and dendritic cells; suggesting that siLP Tregs may still receive MHCII-independent signals from APCs, perhaps via costimulation molecules and/or paracrine cytokine stimulation. Indeed a short-term costimulation blockade with CTLA4-Ig reduces Nur77 expression on K14 siLP Tregs.
Taken together, these results suggest that the intestinal microenvironment uniquely maintains the differentiation and homeostasis of effector Tregs independent of MHCII-TCR interactions.
B1 cell IgE impedes B2 cell IgE-mediated parasite expulsion
Authors: Martin Rebecca , Damle Sheela R. , Valentine Yolander A. , Luker Andrea J. , Lownik Joseph C. , Urban Joseph F. , Conrad Daniel H. 
Published: January 2023

Abstract
Helminth infection is known for generating large amounts of poly-specific IgE. Previously we reported that innate-like B1 cells are responsible for poly-specific IgE production during infection with the nematode parasites Nippostrongylus brasiliensis and Heligmosomoides polygyrus bakeri. We showed that B1 cells make large amounts of IgE. In vitro analysis of factors critical for B1 cell immunoglobulin class switch recombination to IgE demonstrated a requirement for anti-CD40 and IL-4 that further enhanced IgE production when IL-5 was added to culture or when the B1 source was helminth infected mice. We additionally reported that there was also a novel IL-25-induced upregulation of IgE in B1 cells isolated from helminth infected mice. Here, we report that in T cell reconstituted RAG1−/− mice further reconstituted with or without B1 cells and/or B2 cells and followed by infection with N. brasiliensis only B2-reconstituted mice had significantly reduced fecal egg production that was IgE dependent. Parasite clearance was impeded by reconstitution with B1 cells and was also IgE dependent. Helminth-induced B1 cell IgE was not specific for N. brasiliensis because it blocked mast cell degranulation to N. brasiliensis excretory secretory antigens in infected mice. The data supports a hypothesis that B1 cell IgE secretion has evolved as a protective response used by the helminth. In searching for a human B cell equivalent, we additionally identified a novel population of human tonsilar B cells that respond to IL-25 stimulation with increased proliferation and IgE production. We hypothesize that these cells may be important in generation of human poly-specific IgE.
Distinct mechanisms govern resident memory T cell differentiation and survival in different tissues
Authors: Wynne-Jones Erica , Freestone David , Christo Susan N , Yang Kun , Reddiex Scott JJ , Pellicci Daniel G , Carbone Francis R , Kallies Axel , Mackay Laura K 
Published: January 2023

Abstract
Tissue-resident memory T (Trm) cells are a recently defined population of memory T cells that persist at the site of previous infection and contribute to protective local immunity. Trm cells have been identified in both epithelial and solid organs; however, despite their ubiquity, they share molecular commonalities that are distinct from circulating memory T cells. Therefore, we sought to explore the transcriptional regulation of Trm cells in a number of different organs. Our earlier data demonstrated that Trm cells share a core transcriptional signature regulated by the two related transcription factors Hobit and Blimp1. Here we show that Trm cells also have distinct molecular requirements according to anatomic location. While Hobit was uniquely required for the development of Trm cells in all organs tested, Blimp1 was required in some organs, but dispensable in others. Similar to Blimp1, the T-box transcription factor T-bet was also required for Trm cell development in only a subset of organs. This differential transcription factor dependency was linked to distinct cytokine requirements for Trm cell development in different organs. Trm cell defects were enhanced in the absence of multiple transcription factors, suggesting functional redundancy and implicating a role for distinct transcriptional networks as drivers of Trm cell development in different organs. Together, these data demonstrate the adaptation of Trm cells to specific tissue microenvironments and highlight the importance of studying these cells in a variety of organs.
Inflammasome Independent Leukotriene B4 Production Drives Crystalline Silica Induced Sterile Inflammation in Lungs
Authors: Hegde Bindu , Satpathy Shuchismita , Bodduluri Sobha Rani , Jala Venkatakrishna R , Bodduluri Haribabu 
Published: January 2023

Abstract
Occupational exposure to Crystalline Silica (CS) is a major public health concern with millions of workers continuously exposed to CS. Deposition of fine silica particles in the lungs can lead to severe lung inflammation and cancer. Lipid chemokine Leukotriene B4 (LTB4) and IL-1b that mediate leukocyte recruitment to the site of injury, are the key mediators of CS-induced sterile inflammation. Our previous studies have shown that CS exposure increases lung tumor burden in K-rasLA1 mice and that is abrogated in the LTB4 receptor, BLT1 deficient mice. Here, we show that phagocytosis of CS sequentially activates a series of pathways leading to LTB4, IL-1b and CXC chemokine production. CS uptake induces rapid formation of lipid bodies termed “lipidosome” independent of inflammasome activation. Studies with inhibitors suggested that CS-induced lipidosome formation and LTB4 production is linked to the phagocytosis pathway, an observation confirmed by confocal microscopy. We further demonstrated the SiRNA knockdown of cPla2, JNK or Rab40c prevents the formation of lipidosome and production of LTB4. Lastly, we demonstrate using an air pouch model in mice that JNK inhibitor, BI-78D3 is effective in reducing CS induced sterile-inflammation in-vivo. In summary, our studies highlight a novel inflammasome independent and JNK activation dependent lipidosome pathway leading to the production LTB4 as a major regulator of CS-induced sterile inflammation.