bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel approaches to assess therapeutic efficacy in the MRL/lpr model of lupus
Authors: Schaevitz Laura , Lim Maria , Defensor Erwin 
Published: January 2023

Abstract
Spontaneous genetic models of systemic lupus erythematosus (SLE), such as MRL/lpr and NZB/W F1 mice, are commonly used to assess therapeutic efficacy. Therapies are evaluated against several reliable standard measures, including proteinuria, anti-dsDNA, skin lesions, splenomegaly, nephritis, and survival. We hypothesize that continuous monitoring of behavioral and physiologic parameters will provide additional meaningful data to assess disease and efficacy. Here, we treated MRL/lpr mice with two reference compounds, cyclophosphamide (CP: 25mg/kg, IP, weekly) and dexamethasone (DEX: 2mg/kg, IP, 3× week). Throughout the experiment, mice were housed in Vium’s Digital Vivarium, which provides continuous monitoring and assessment of behavioral and physiologic measures using a network of cameras and sensors. Digital metrics and standard measures were displayed and analyzed in near real-time, through an online interface. Group disease profiles were compared using automatically generated breathing rate and motion metrics, in addition to standard measures. Proteinuria, body weight, and spleen size showed disease establishment in MRL/lpr mice as early as 16 weeks of age, which was ameliorated by CP or DEX (p’s &lt; 0.03). Breathing rate significantly increased in MRL/lpr vehicle-treated mice as early as 13 weeks of age, while nighttime motion significantly decreased in MRL/lpr vehicle-treated mice by 14 weeks. Changes in breathing rate and motion detected in MRL/lpr mice were rescued by administration of CP or DEX (p’s &lt; 0.0001). Our results demonstrate that continuous measurement of breathing rate and motion are useful additional measures for evaluating disease severity and therapeutic efficacy in the MRL/lpr murine model of SLE.
A Novel Clone Ranking and Selection Assay that Combines all the Relevant Insights Needed to Choose the Most Optimal Clones in a Single Experiment
Authors: Duensing Thomas , Liu Zhaoping , Hauser Caroline 
Published: January 2023

Abstract
When developing cell lines for the production of antibodies, it is essential to choose healthy clones with high levels of IgG per cell. This process typically requires several assays on multiple platforms to get enough information to select optimal clones. Due to the expense and effort involved, this limits the number of clones that can be evaluated. We have developed the Cy-Clone™ PLUS Kit, a rapid, no-wash screening assay that simultaneously provides IgG concentration, IgG concentration per cell and per viable cell, cell viability, and cell density in every well of a 384-well plate.
CHO cells transfected with human IgG genes were inoculated, plated by limiting dilution, and cultured for 20 days. Cells were mixed with the reagents from the Cy-Clone PLUS kit and then read on the iQue® Screener PLUS platform in ~22 minutes. Using ForeCyt® Software, readouts for each well were visualized using heat maps and Profile Map to identify wells meeting multiple IgG production criteria.
The Cy-Clone PLUS kit with the iQue Screener platform provides all the relevant data needed to select and rank the best clones in a single well without screen-size limitations. The Cy-Clone PLUS kit enables you to make more informed decisions earlier, shorten development time, and decrease costs.
Central immune tolerance establishment under thymic involution (Supported by NIH/NIAID 1R01AI121147 to D-M Su)
Authors: Su Dongming , Oh Jiyoung , Wang Weikan , Thomas Rachel 
Published: January 2023

Abstract
The establishment of central immune tolerance in the thymus is occurs via thymocyte negative selection to deplete self-reactive T clones and thymic CD4+ regulatory T cell (tTreg) generation. Due to limited published data, it is largely unclear whether these two mechanisms are both defective in the involuted thymus, as a result of aging or other insults. We used mouse models with thymic epithelial cell (TEC) conditional defect (FoxN1 conditional knockout) and naturally aged defect to study mechanistic changes in central tolerance establishment. We identified a defect in thymocyte negative selection but not in tTreg generation. In comparison to tTreg with thymic conventional CD4+ T cells (tTcon) in the same thymus, the ratios were increased with severity of thymic involution, suggesting that activity of tTreg generation is increased in the involuted condition. We found that these changes were due to alterations in signaling strength received by T cell receptors, characterized by a decline of Nur77 and CD5 in CD4+ single positive thymocytes and an enhancement of Zap-70 in newly-generated tTregs. This is due to weakened signaling provided by MHC/self-peptide complex ligand from defective TECs, but not due to changes in tTreg development-required factors (TGFb, IL-2, IL-15, and NF-kB). Our results suggest that the involuted thymus attempts to balance the defective negative selection by enhancing tTreg generation to maintain central immune tolerance. If this balance is broken, age-related inflammatory diseases could take place.
CD46 activation on monocyte-derived dendritic cells
Authors: Biltoft Mette , Hansen Aida S. , Höllsberg Per 
Published: January 2023

Abstract

Objective
CD46 is an important immune modulatory receptor that influences the expression of certain cytokines in different cell types. This function has been proposed to depend on expression of specific isoforms of CD46. The common isoforms are named BC1, BC2, C1 and C2, and activation of T cells induces a switch towards dominance of BC2 and C2. However, the effect of CD46 activation on the cytokine production in dendritic cells (DCs) has so far been inconclusive and the relative distribution of CD46 isoforms in monocytes and DCs is unknown.

Methods
Monocytes were isolated from peripheral mononuclear blood cells and differentiated to dendritic cells (moDC). The maturation of moDCs was confirmed by flow cytometry using a panel of maturation markers. Monocytes and moDCs were stimulated with LPS in presence or absence of CD46 activation and the production of various cytokines were analyzed by real-time PCR. The monocytes and moDCs were furthermore analyzed for their expression of different CD46 isoforms by an isoform-specific real-time PCR assay.

Results
Upregulation of CD40, CD80, and CD83 on the surface of moDCs demonstrated the maturation of monocytes to moDCs. Activation of CD46 in moDCs during LPS stimulation was found to reduce the mRNA expression of IL12p40 and CXCL10 while no effect of CD46 activation on these cytokines was observed in monocytes. Examination of CD46 isoforms revealed a dominant expression of the isoforms BC2/C2 in moDCs upon differentiation from monocytes.

Conclusion
The results presented here suggest that CD46 activation reduces the pro-inflammatory potential of LPS stimulation in moDCs but not in monocytes. These functional changes are associated with a switch towards dominant expression of BC2/C2 isoforms.
Using molecular chemokine adjuvants to target the germinal center reaction and enhance HIV-specific humoral responses to DNA immunogens in the mucosa or the periphery
Authors: Gary Ebony Nicole , Bernui Mariana , Weiner David , Kutzler Michele A. 
Published: January 2023

Abstract
The germinal center (GC) is a dynamic cellular substructure found in lymphoid organs where follicular helper T cells (TFH) provide the necessary proliferation stimuli and cytokine signaling that promote antibody classs-witching and affinity maturation of B cell receptors. DNA vaccines have demonstrated great potential as vaccine immunogens and in vivo electroporation (EP) has enhanced this immunogenicity. The ability to deliver plasmid encoded molecular adjuvants to direct immune responses is an advantage of the DNA platform. Mucosa-associated epithelial chemokine (MEC, or CCL28) is secreted by epithelial cells that line mucosal surfaces, it signals through CC-chemokine receptors 10 and 3 (CCR10 and CCR3) which are expressed on IgA+ plasma cells, TH17 cells, and mucosa-resident antigen presenting cells. IL-12 is a T cell stimulating cytokine that promotes a helper-T cell phenotype. Mice were vaccinated twice intramuscularly with either consensus HIV-1 env DNA alone or DNA plus molecular MEC or IL-12 delivered by in vivo EP. Fourteen days post vaccination, animals were sacrificed and spleens and draining lymph nodes were harvested for flow cytometric analysis. Both MEC and IL-12 significantly increased antigen-specific GC B cells (B220+CD138−Igm−IgD−PNA+CXCR5+gp120+) (11.1% and 8.91% respectively, compared to 6% with DNA alone). Antigen specific isotype- ELISA confirmed that MEC increases IgA in serum (900ng/ml compare to 600ng/ml with DNA alone) and fecal extracts (27ng/ml compared to 22ng/ml), while IL-12 increases IgG in serum alone (2200ng/ml compared to 1300ng/ml with DNA alone). These data indicate that molecular adjuvants can be used to specifically direct immune responses to the mucosa or the periphery.
Dengue NS1-specific IgY antibodies neutralizes dengue infection without inducing antibody dependent enhancement.
Authors: O’Donnell Kyle Lee , Fink Ashley , Nilles Matthew L. , Bradley David S 
Published: January 2023

Abstract
Dengue virus (DENV) is the most prevalent arbovirus with worldwide cases increasing from 2.2 to 3.2 million in the last 5 years. Symptoms of DENV infection range from a mild fever to extensive vasculature permeability resulting in hemorrhagic fever and shock. Passive and active vaccines have been unsuccessful in protecting from or treatment for DENV infection because of antibody dependent enhancement (ADE), increased viral load due to increased monocyte opsonization by non-neutralizing Abs from a different DENV serotype. We have previously demonstrated that polyclonal avian IgY generated against whole, killed DENV-2 ameliorates DENV infection in mice without ADE. The lack of ADE with therapeutic IgY supported our hypothesis that IgY may provide a viable treatment for DENV infection as mammalian Fc receptors are unable to bind to IgY. Here we isolated DENV epitope-specific IgY and test these purified IgY pools for their ability to neutralize DENV in vitro. Avian IgY specific for Non-Structural-1 (NS1) demonstrated neutralization capacity in vitro equal to the whole polyclonal DENV IgY preparation. DENV-NS3 specific IgY, on the other hand, did not demonstrate any neutralization of DENV in vitro. We propose that NS1-specific IgY may provide a viable oligoclonal treatment for DENV infection in humans without inducing ADE.
Loss of IL-4Rα signaling is partially able to restore anti-Mycoplasma pneumoniae IgG titers during allergic airway disease
Authors: Totten Arthur H , Xiao Li , Luo Danlin , Crabb Donna M , Ratliff Amy E , Waites Ken B , Atkinson T. Prescott 
Published: January 2023

Abstract

Rationale
Mycoplasma pneumoniae (Mpn) is the leading bacterial agent of pediatric community acquired pneumonia. Epidemiologic evidence has suggested increased susceptibility to Mpn infection among asthmatics. We hypothesized that allergic airway sensitization would impair host immune responses to Mpn infection, but that loss of Th2 cytokine signals (IL-4, IL-13) would overcome this impairment in a murine model.

Methods
Wild type (Wt) and IL-4Rα−/− BALB/cJ mice were sensitized and challenged with ovalbumin (OVA) to induce allergic airway inflammation, and then infected with Mpn. Immune parameters were studied by analysis of cellular profiles in bronchoalveolar lavage fluid (BALF), serum IgG and IgE antibody levels to a whole bacterial antigen preparation, recombinant Mpn P1 adhesin (rP1), and OVA. Total lung RNA was used to examine cytokine transcript levels for Th1, Th2 and Th17 biasing transcripts.

Results
Anti-Mpn and P1-specific total IgG responses were decreased in allergen-sensitized animals compared to unsensitized controls (p &lt; 0.01). Decreased titers of IgG1, IgG2a, IgG2b and IgG3 were present in rP1-specific, but not OVA-specific IgG subclass levels. Loss of IL-4Rα signaling partially restores total and subclass-specific responses. Th2 cytokine mRNA levels were increased in Wt, but not IL-4Rα−/− OVA sensitized groups compared to controls. IL-4Rα−/− animals had increased Interferon-γ(IFN-γ) and IL-17A mRNA levels compared to Wt animals.

Conclusions
Anti-Mpn IgG antibody titers were decreased in Wt allergic animals compared to unsensitized controls and the impairment in immune response to the pathogen was partially restored by inhibition of Th2 signaling in IL-4Rα−/− animals.
The distinct role of Tfh and Th17 cells and their interplay in the pathogenesis of rheumatoid arthritis
Authors: Zhang Xin , Migliore Frank , Budziakowska Magdalena , Quresh Quretul , Zakem Jerald , Webb-Detiege Tamika , Davis Willam , You Zongbing , Quinet Robert 
Published: January 2023

Abstract

Objectives
Rheumatoid Arthritis (RA) is an autoimmune disease characterized by the presence of autoantibodies and chronic inflammation in synovium leading to joint destruction. Recently Tfh and Th17 cells have emerged as novel T cell subsets controlling autoimmunity. Our previous data showed that circulating Tfh (cTfh) and circulating Th17 (cTh17) cells were increased in RA patients. Here, we further investigated their role and interplay in the pathogenesis in RA.

Design and Methods
Peripheral blood was collected from 61 RA patients and 61 healthy donors. RA patients were divided into remission (&lt;2.6) and active groups (&gt;2.6) based on disease activity score. Clinic parameters including RF, anti-CCP, ESR and CRP were obtained. The frequencies of cTfh and cTh17 cells were measured by flow cytometry. Serum IL-21 and IL-17 were detected by ELISA. Correlations of the frequency of cTfh/IL-21 with cTh17/IL-17 and clinic parameters were statistically determined.

Results
The increased frequency of cTfh cells significantly correlated with the level of anti-CCP antibody (p&lt;0.01) in active RA patients, whereas increased frequency of cTh17 cells correlated with CRP level (p&lt;0.05). Furthermore, the level of cTfh cells and serum IL-21 level were significantly correlated with the percentage of cTh17 cells in RA patients (P&lt;0.05). The serum IL-17 level only closely correlated with CRP in RA patients.

Conclusion
Tfh cells may be involved in RA pathogenesis by inducing generation of autoantibody producing plasma cells, while cTh17 cells largely related to inflammation. Tfh cells and its cytokine IL-21 may also steer T cell differentiation into Th17 cells which secrete the pro-inflammatory cytokine IL-17 and contribute to inflammation in RA.
Role of SIRT1 in C5b-9 mediated oligodendrocyte cell cycle activation
Authors: Tatomir Alexandru , Rao Gautam , Boodhoo Dallas , Ciriello Jonathan , Rus Violeta , Rus Horea 
Published: January 2023

Abstract
SIRT1 is a NAD-dependent histone deacetylase involved in the regulation of transcription, apoptosis, metabolism and differentiation. We have previously demonstrated that sublytic levels of complement C5b-9 terminal complex increased the survival of oligodendrocytes (OLGs) and induced their dedifferentiation. In this study we investigated the role of SIRT1 in OLGs differentiation and the effect of sublytic levels of C5b-9 on SIRT1 expression. We also investigated the downstream effects of SIRT1 by measuring histone H3 Lysine 9 trimethylation (H3K9me3) and expression of cyclin D1. OLG progenitor cells purified from the brain of rat pups were differentiated in vitro and stimulated with sublytic C5b-9 or C5b6 for 3, 6 and 18 h. The level of SIRT1 mRNA was measured using real-time PCR and SIRT1, cyclin D1 and H3K9me3 protein expression were measured using western blotting. We found a decreased expression of SIRT1 mRNA and protein, a decreased level of H3K9me3 and an increased expression of cyclin D1 during OLG differentiation. Stimulation of OLGs with sublytic C5b-9 for 3h resulted in a significant decrease in SIRT1 mRNA and protein levels while stimulation with C5b6 had no effect. SIRT1 protein level in OLGs after 8h of exposure to C5b-9 were significantly lower than in C5b6 treated cells. H3K9me3 levels also decreased significantly after stimulation with C5b-9 as compared with unstimulated or C5b6-treated OLGs. Cyclin D1 expression increased after stimulation with C5b-9, indicating cell cycle activation. Our data show that C5b-9 stimulation of OLGs reduces SIRT1 expression, contributing to cell cycle activation by decreasing repressive trimethylation of histone H3 lysine 9.
Expression of the TNF-family cytokine TL1A (TNFSF15) by renal CX3CR1+ resident mononuclear phagocytic cells and role of TL1A in autoimmune nephritis
Authors: Richoz Nathan , Farley Taylor K. , Chalmers Samantha A. , Putterman Chaim , Meylan Francoise , Clatworthy Menna R. , Siegel Richard M. 
Published: January 2023

Abstract
Systemic lupus erythematosus is a polygenic autoimmune disease with a broad spectrum of clinical manifestations including organ-threatening kidney inflammation associated with immune complex (IC) deposition. The kidney is populated by a rich network of tissue-resident mononuclear phagocytes that express Fc receptors (FcRs) and internalize ICs. TL1A (TNFSF15) is a tumor necrosis factor (TNF) superfamily cytokine that can be induced in myeloid cells following ligation of TLR’s and FcRs by IgG IC’s and co-stimulates T cells through its receptor DR3. We have found that DR3 deficiency protects MRL/lpr lupus-prone mice from the renal inflammation but not systemic autoimmunity that develops in these mice, suggesting that TL1A/DR3 interactions could play a role in activating autoimmune T cells and antigen presentation in the kidney. We identified four distinct populations of renal phagocytes based on the expression of F4/80 and CD11b. One population of cells constitutively expressed high levels of surface class II and the CD28 ligand B7-2(CD86), suggesting that they would be potent antigen presenting cells. A fifth population expressing similar markers was present only in inflamed kidneys, both in the MRL/lpr mouse model of lupus and nephrotoxic serum-induced nephritis. Both of these populations constitutively expressed surface TL1A. The new population increases with the severity of nephritis and is unequally distributed between the cortex and the medulla. We will further analyze these populations via transcriptomics and functional assays after immune complex stimulation. This work identifies CX3CR1+ MNP’s as possible antigen-presenting cells in the kidney and a source of TL1A for costimulation of autoreactive T cells in nephritis.
CXCL17 is a stomach-expressed chemokine and plays a role in diet-induced obesity
Authors: Hernandez Marcela , Herrera Carolina , Nguyen Hong-Tam , Zlotnik Albert 
Published: January 2023

Abstract
CXCL17 is a mucosal chemokine that attracts dendritic cells and monocytes to specific tissues. CXCL17 is mainly expressed in the respiratory and gastrointestinal tracts and has been shown to have anti-inflammatory effects. Its recently described receptor, GPR35/CXCR8, is a GPCR that has been involved in metabolic functions. Given that macrophages are known to be involved in fat tissue homeostasis, we decided to study a possible role of the CXCL17/CXCR8 axis in obesity. To this end, we used a mouse model of diet-induced obesity to compare Cxcl17−/− mice and wild type (WT) C57BL/6 mice. Mice were fed either a 10% or 60% fat diet for four months. Their development and weight gain as well as several metabolism tests were monitored for two or four months. Plasma and fat tissue samples were used in adipokine assays and to measure levels of obesity-related cytokines and hormones. Gastrointestinal tract tissue, adipose and liver tissues were obtained to perform further RNA or protein assays.
The results indicate that Cxcl17−/− mice fed a 60% fat diet gained significantly more weight than WT mice. We also determined that under homeostatic conditions CXCL17 is highly expressed in the upper gastrointestinal tract (oral cavity, esophagus and stomach) in WT mice. Obese Cxcl17−/− mice also exhibit impaired response to glucose in an insulin tolerance test. Adipokine assay results revealed differences in FGF-21 levels and in other obesity-associated molecules between WT and Cxcl17−/− mice. We conclude that CXCL17 influences development of obesity and systemic metabolic dysfunction. Our results suggest that CXCL17 may be a therapeutic target to treat obesity-related conditions.
Structures of gasdermin-D C-terminal domain reveal mechanisms of autoinhibition
Authors: Liu Zhonghua , Wang Chuanping , Yang Jie , Rathkey Joseph , Abbott Derek W , Xiao Tsan Sam 
Published: January 2023

Abstract
Gasdermin-D (GSDMD) is a member of the gasdermin family, and is a critical mediator for host defense against microbial infection and danger signals. Inflammatory caspases (caspases-1, 4, 5, and 11) cleave GSDMD to generate an N-terminal cleavage fragment (GSDMD-N) that oligomerizes in cell membrane to form ring-like pores during pyroptosis, an informatory form of programmed cell death. In addition, the GSDMD membrane pores facilitate the release of inflammatory cytokines such as IL-1b and IL-18. However, how GSDMD-N is inhibited by the C-terminal domain of GSDMD (GSDMD-C) in the rest state is unknown. In addition, the mechanisms for the release of GSDMD-N from GSDMD-C upon caspase cleavage and the formation of membrane pores have not been established. Here we report the crystal structures of the C-terminal domains of human and murine GSDMD protein. Even though the overall structures of the two GSDMD-C structures are similar to that from mGSDMA3, two regions of the helical structures in GSDMD-C are significantly different from that in mGSDMA3. Overexpression of GSDMD-N induces lysis of the bacterial expression host, while concomitant expression of GSDMD-C reduces such bactericidal activities. Mutation of GSDMD-C residues involved in autoinhibition attenuates the intramolecular domain-domain interactions, as well as enhances pyroptosis in 293T cells transiently expressing GSDMD. Our study thus reveals mechanisms of autoinhibition of GSDMD, and suggests potential therapeutic application of GSDMD as a bactericidal agent.
Attenuation of Osteoclast Differentiation by the Knockdown of Nicotinamide PhosphoribosylTransferase
Authors: Islam M Shamima , Heruth Daniel P , Zhang Li Qin , Ye Shui Qing 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is characterized by chronic inflammation and progressive bone destruction. Nicotinamide phosphoribosyltransferase (Nampt) functions in NAD synthesis, inflammation and apoptosis. Our recent finding that Nampt+/− mice were protected against bone erosion in a Collagen Induced Arthritis mouse model led us to hypothesize that Nampt may play a critical role in osteoclast differentiation and knockdown of Nampt could attenuate osteoclast formation.
We studied the effects of decreased Nampt expression in osteoclastogenesis in both Nampt siRNA transfected RAW macrophages and bone marrow macrophages (BMM) from wild-type (Nampt+/+) and Nampt+/− mice. Number of differentiated osteoclasts were decreased from Nampt deficient RAW cells (86±2.8% vs 100%, N=3, p&lt;0.001) and Nampt+/− BMMs (13.4±3 vs 17±4.8%, N=3, p=0.02 by paired T-Test) vs Nampt+/+ controls. Acp5 (0.1±0.035, N=4, p&lt;0.0001), and Nuclear factor of activated T-cells (Nfatc1) mRNA were all decreased (0.29±0.08, N=4, p&lt;0.0001) in Nampt deficient cells relative to the scrambled control. Luciferase reporter activity was decreased in RAW cells co-transfected with Nfatc1 luciferase reporter and Nampt siRNA relative to the scrambled control (0.33±0.18, N=3, p&lt;0.005). ChIP analysis demonstrated that the epigenetic structure of the Nfatc1 promoter was altered in response to decreased Nampt with decreased histone acetylation.
In summary, our data suggest that knockdown of Nampt transcriptionally inhibited Nfatc1 expression via decreasing histone acetylation, resulting in an attenuated osteoclastogenesis. Nampt could be a potential new risk factor and therapeutic target for arthritis.