bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nuclear Factor-κB is a Potential Therapeutic Target in Cystitis (142.9)
Authors: Vykhovanets Eugene , Vykhovanets Olena , Ponsky Lee , Cherullo Edward , Gupta Sanjay 
Published: January 2023

Abstract
NF-κB has been shown to be activated in various inflammatory disorders or by physical and chemical stress. NF-κB regulates the transcriptional of genes important for chronic inflammation and drug resistance. Interstitial cystitis (IC) affects millions of women and is characterized by severe pain and chronic inflammation. Etiology and the mechanisms involved in pathogenesis of IC are still a matter of obscure. Animal models of IC and role of NF-κB as a therapeutic target in the IC have not been elucidated. Using transgenic NF-κB-Luciferase Tag female mice, we performed serial, noninvasive in vivo and ex vivo molecular imaging of NF-κB activation in the mouse body after administration of arsenic trioxide (5 mg/kg), LPS (2 mg/kg), or cyclophosphamide (CYC, 200 mg/kg) to initiate transient cystitis. Pretreatment with dexamethasone (DEX) (10 mg/kg) was used to modulate the chemical-induced NF-κB-dependent luminescence. Treatment of NF-κB-Luc mice with chemicals increased luminescence in a time- and organ-specific manner. Highest levels of bladder NF-κB-dependent luminescence were observed 4 hr after CYC administration. Pretreatment with DEX 1 hr before injection of CYC significantly down-regulated CYC-induce bladder NF-κB-dependent luminescence and gross anatomy signs of cystitis. The molecular imaging of NF-κB activity might be an attractive approach to distinguish the role of NF-κB signaling in transient cystitis and select potential candidate drugs for treatment of the IC.
Intricate regulation of IL-17 response by IFN-γ and IL-27 determines the outcome of the pathogenetic events in experimental autoimmune arthritis (143.18)
Authors: Rajaiah Rajesh , Puttabyatappa Muraly , Moudgil Kamal 
Published: January 2023

Abstract
Recent advances in defining the new subsets of T cells and the related newer cytokines have stimulated new undertakings in examining the pathogenesis of autoimmunity beyond the traditional Th1-Th2 paradigm. We report here novel findings regarding the roles of IFN-γ, IL-17 and IL-27 in the adjuvant arthritis (AA) model of human rheumatoid arthritis that challenge the traditional viewpoints. We hypothesized that the temporal kinetics and the relative levels of antigen-induced cytokine responses determined both the propagation of AA as well as resistance to the disease. We determined the cytokine responses of arthritic Lewis rats at different phases of the disease and that of the arthritis-resistant, MHC-compatible Wistar Kyoto (WKY) rats given a potentially arthritogenic challenge. In Lewis rats, IL-17 dominated the early (incubation phase) response, whereas IFN-γ and IL-27 appeared late (peak phase). Intriguingly, in WKY rats, all these 3 cytokines appeared concomitantly and early, showing that IFN-γ and IL-27 successfully pre-empted the pathogenicity of IL-17. This cross-regulation, along with the novel finding of upregulation of IL-27 by IFN-γ, was validated through mechanistic experiments, including the AA-protective effect of IFN-γ injected in vivo, and the differential modulation of specific signaling biomolecules and inflammatory mediators in the joints. These results provide a novel IL-17-IL-27-centered perspective on the pathogenesis of autoimmune arthritis.
IgD BCR-mediated signals do not have protective roles in negative selection of autoreactive B cells in lpr mice (143.29)
Authors: Guo Linjie , Guo Zhuxiu , Zheng Biao , Han Shuhua 
Published: January 2023

Abstract
IgD is the major antigen receptor isotype coexpressed with IgM on the surface of most peripheral B cells in mice and humans. However, the biological role of IgD as B cell receptor (BCR) has remained enigmatic and controversial. Previous studies have indicated that IgD may play a role in B cell tolerance and the expression of IgD may rescue cells undergo programmed cell death. To understand the role of IgD in B cell tolerance and autoimmunity, we have examined the development of autoimmune syndrome in lpr mice deficient for IgD. The present study showed that IgD deficiency did not alter many aspects of the autoimmunity in lpr mice such as splenomegaly and significant expansion of activated T cells. Interestingly, lpr mice with IgD deficiency exhibited increased levels of autoantibodies and more severe nephritis compared to their IgD-competent littermates. In addition, the absence of IgD did not result in any increase in cell death of B cell subsets including transitional B cells. Thus, our results indicated that IgD BCR-mediated signals do not have a protective role in negative selection of autoreactive B cell clones. In contrast, the absence of IgD leads to heightened autoantibody production and more deposition of immune complex in the kidney, resulting in more severe nephritis.
Loss of P2X7 receptor membrane expression in B220+ double-negative T lymphocytes of autoimmune MRL/lpr mice (143.32)
Authors: Bobé Pierre , Le Gall Sylvain , Legrand Julie , Benbijja Mohcine , Benihoud Karim , Kanellopoulos Jean 
Published: January 2023

Abstract
Lupus is an autoimmune disease influenced by multiple genetic loci. Fas-deficient MRL/lpr mice exhibit lupus and lymphoproliferative syndromes with massive accumulation of B220+CD4-CD8- (DN) T lymphocytes. Pathogenic B220+ DN T lymphocytes are derived from normal activated T lymphocytes which have not undergone activation-induced cell death due to inactivation of Fas. Normal activated peripheral T lymphocytes up-regulate the membrane phosphatase B220 before undergoing apoptosis. In Fas-deficient mice, activated T cells down-regulate CD4 or CD8 molecules but retain the expression of B220. P2X7R is an extracellular ATP-gated cell membrane receptor involved in the release of inflammatory cytokines and cell death. We show herein that MRL/lpr mice carry a P2X7R allele, which confers a high sensitivity to ATP. However, during aging, the MRL/lpr T-cell population exhibits a drastically reduced sensitivity to ATP-mediated stimulation of P2X7R. This decrease in P2X7R activity parallels the increase in B220+ DN T-cell numbers in lymphoid organs. The rare B220+ T cells observed in normal mice are also resistant to ATP treatment. Moreover, B cells, which express B220 as a developmental marker, are also dramatically less sensitive to ATP treatment. Using rabbit anti-P2X7R sera, we found a strong reduction in P2X7R levels on B220+ cells. Our results prompt the conclusion that B220 expression strongly correlates with the negative regulation of ATP/P2X7R pathway on both T and B cells.
REGULATORY CD4-CD8- T CELLS PREVENT AUTOIMMUNE DIABETES (143.4)
Authors: Lesage Sylvie , Dugas Véronique , Hillhouse Erin , Beauchamp Claudine , Chabot-Roy Geneviève 
Published: January 2023

Abstract
Regulatory T cells show great potential for use in cellular therapy. In particular, CD4-CD8- (DN) T cells, which compose 1 to 3% of total T lymphocytes, exhibit prominent antigen-specific immune tolerance properties in models of allografts and xenografts, as well as in an induced model of autoimmune diabetes. Here, we evaluate the immunoregulatory properties of DN T cells in the autoimmune-prone NOD genetic background. Using the 3A9 TCR transgenic mice, we demonstrate that DN T cells from both autoimmune-resistant and -prone mice are equally effective at eliminating potentially autoreactive B cells in vitro. However, autoimmune-prone mice carry at least three fold fewer DN T cells than autoimmune-resistant mice, in both TCR transgenic and non-transgenic setting. Interestingly, a single transfer of DN T cells is sufficient to prevent autoimmune diabetes onset in autoimmune-prone mice. These results suggest that increasing DN T cell number is sufficient to confer protection from autoimmune diabetes onset. Further functional and genetic characterization of DN T cells in NOD mice highlighted a potential role for both CD172a and IL-10 in the regulation of DN T cell homeostasis. Importantly, genetic polymorphisms in both CD172a and IL-10 are associated with diabetes susceptibility in humans. Taken together, our results strongly suggest a role for DN T cell in peripheral tolerance and the association of genetic defects in DN T cell homeostasis with the development of diabetes.
IL-13 Mediated Regulation of Islet-Reactive T Cells (143.44)
Authors: Rasche Sarah , Phillips Michele , Quinn Anthony 
Published: January 2023

Abstract
Immunotherapy with the beta cell antigen glutamic acid decarboxylase (GAD65) or GAD65 peptides delays the onset of type 1 diabetes (T1D) in NOD mice, while alum formulated rGAD65 may be therapeutic in human T1D; nevertheless, the regulatory mechanisms involved in each therapy may be distinct. Here, we sought to discover the factors that contribute to GAD65 peptide 524-538 (p524)-mediated protection from T1D in NOD mice and to determine if IL-13 produced by p524-specific T cells could regulate pathogenic islet-reactive T cells. IL-13 produced by p524-reactive T cells directly modulated IFN-g production by spontaneously arising GAD65 530-543-specific Th1 cells in NOD mice and rIL-13 inhibited proliferative responses of islet-reactive BDC2.5 clonotypic T cells. Contrary to previous observations, both CD4+ and CD8+ T cells in the peripheral lymphoid compartment of NOD and C57BL/6 mice expressed the receptor subunit required for IL-13 signaling, IL-13Rα1. Interestingly, the number of IL13Rα1+ T cells was highest in pancreatic lymph nodes, as compared to inguinal lymph node or splenic T cells, and this expression was uniquely diminished in pancreatic lymph nodes of older prediabetic NOD. Thus, IL-13 may be an important mechanism for regulation of autoreactive T cells in the early stages of insulitis. The loss of IL-13Rα1 expression on T cells in the draining nodes of the pancreas may reflect fading regional immune regulation and progression from insulitis to overt T1D.
The paradoxical role of Th1 effector CD4 T cells during autoimmune chronic inflammation. (143.47)
Authors: Harrington Laurie , Kress Robert , Mollo Sarah , Yeh Wen-I 
Published: January 2023

Abstract
Many autoimmune chronic inflammatory diseases, including multiple sclerosis, have been previously reported as Th1-mediated disorders; however recent data suggests that Th17 cells are responsible for disease. Paradoxically, the principal Th1 cytokine IFNγ does not appear necessary for disease, but the key Th1-associated transcription factor Tbet is required. This conundrum propelled us to investigate the regulation of this transcription factor and effector CD4 T cell functions, as well as the role of Th1 cells during autoimmunity. Following the onset of experimental autoimmune encephalomyelitis (EAE), we observed a preferential upregulation of Tbet by effector CD4 T cells within the CNS, but not the secondary lymphoid organs. These Tbet+ CD4 T cells were capable of producing IFNγ, and a fraction were able to make both IFNγ and IL-17. In contrast, CD4 T cells producing only IL-17 were primarily low for Tbet expression, in keeping with a non-essential role for Tbet during Th17 differentiation. We next examined the requirement of the Th1 promoting cytokines IFNγ and IL-12 for the induction of Tbet+ CD4 T cells during EAE. In the absence of these cytokines, a significant population of CD4 T cells still upregulated Tbet. Consistent with these data, we found that CD4 T cells from IFNγ reporter/IFNγ deficient mice with EAE expressed the Thy1.1 reporter molecule, indicating the presence of Th1-like, Tbet+ CD4 T cells even in the absence of the cardinal Th1 cytokine, IFNγ.
Increased formation of anti-insulin B cells combines with defective central and peripheral tolerance checkpoints to contribute to type 1 diabetes liability in NOD mice (143.5)
Authors: Henry Rachel , Kendall Peggy , Thomas James 
Published: January 2023

Abstract
Insulin-reactive autoantibodies precede disease development in type 1 diabetes (T1D)-prone non-obese diabetic (NOD) mice and humans, consistent with a breakdown in B cell tolerance for this key T1D autoantigen. Accelerated disease occurs in B cell receptor (BCR) transgenic NOD mice which have a small population of anti-insulin B cells. Absence of this specificity in closely related BCR transgene results in disease protection, highlighting the importance of B cell specificity for insulin in promoting disease. The genesis and maturation of anti-insulin B lymphocytes were investigated in NOD mice to identify alterations in tolerance checkpoints which underlie T1D progression. In NOD mice, polymorphic anti-insulin germline Vκ genes promote increased formation of anti-insulin B cells in the bone marrow. In contrast to non-autoimmune mice, failure to cull anti-insulin B cells in NOD mice though receptor editing or deletion in the bone marrow as well as at transitional checkpoints in the periphery permits entry of anti-insulin B cells into T1, follicular, and marginal zone B cell compartments. Entry occurs despite the presence of a large competing repertoire. These findings highlight how multiple points at which B cell tolerance is compromised in NOD mice combine to increase risk of T1D. A broad therapeutic window therefore exists during B cell development, within which successful tolerance restoration could provide effective disease intervention.
MicroRNAs: modulators of T cell phenotype in multiple sclerosis (143.57)
Authors: Smith Kristen , Guerau-de-Arellano Mireia , Williams Jessica , Racke Michael , Lovett-Racke Amy , Whitacre Caroline 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an inflammatory disease of the central nervous system mediated by autoreactive CD4+ T cells of a Th1 and Th17 phenotype. Identifying novel factors that control Th differentiation will lead to a better understanding of disease pathogenesis and the development of new therapeutic strategies. MicroRNAs (miRs) are a class of small non-coding RNAs that negatively regulate post-transcriptional gene expression. MiRs have an established role in immune system homeostasis, and profiling studies have found differential expression of miRs in autoimmune diseases, including MS. Dicer-deficient CD4+ T cells that lack miRs produce more cytokine upon stimulation. Together these data indicate the miR(s) are responsible for controlling cytokine expression and act as negative regulators of inflammation. However, the specific miR(s) responsible for modulating inflammatory cytokine production and Th programming have not been defined. Using target prediction algorithms, we identified miRs predicted to target genes that mediate Th1 specification and function. We have validated these miR-target pairs using gain- and loss-of-function experiments in human Th1-polarized cells. We have also profiled the expression of these miRs in specific CD4+ T cell subsets from MS patients. Our results identify miRs that modulate Th developmental programming and effector function and whose expression is altered in MS. These miRs may be useful immunomodulatory treatments or biomarkers in MS.
Over-expression of the transcription factor Bright alters B cell tolerance (143.9)
Authors: Oldham Athenia , Miner Cathrine , Webb Carol 
Published: January 2023

Abstract
The persistence of circulating auto-antibodies is indicative of a breach in B cell tolerance. Mice that constitutively over-expressed the transcription factor Bright throughout B cell development produced antibodies to nuclear antigens (ANAs) similar to those produced in autoimmune diseases such as lupus. Whereas, normal mice reduce Bright expression in mature B cells and do not produce ANAs. ANAs did not result from global hyperglobulinemia and were IgG demonstrating that they originated from B cells that had transited the germinal center. ANA production was dependent upon Bruton's tyrosine kinase. To determine where tolerance breaches occurred in this autoimmune model, we analyzed transgenic B cells for abnormalities at multiple tolerance checkpoints. We found that early transitional B cells responded to BCR stimulation with exaggerated cell death, suggesting the clonal deletion checkpoint was in tact. Transgenic mice also developed normal numbers of naturally occurring anergic B cells. Interestingly, mature B cell numbers were down by half and those cells were hyper-responsive to proliferative signals. The marginal zone compartment is enriched for self reactive B cell. A bias towards marginal zone versus follicular B cell development was evident both in cell numbers and through crosses with Bcl2 transgenics. Together, these data suggest that appropriate regulation of Bright levels is critical for maintaining normal B cell tolerance.
Automated determination of optimal microarray data processing (144.11)
Authors: Scheuermann Richard , Kong Y. Megan , Wang Siling , Qian Yu , McGee Monnie 
Published: January 2023

Abstract
Gene expression microarrays are widely used to study genome-wide gene expression profiles. Many microarray analysis methods are available, making it challenging to decide which method to use. While the effectiveness of some of these methods has been assessed using artificial spike-in data sets, analytical approaches that work well with spike-in data may not work as well with data from real biological samples. To evaluate these methods we applied Gene Ontology (GO) term co-clustering as a comparative tool to evaluate 300 different data processing pipelines composed of various background correction, normalization and summarization methods using real biological data, based on the premise that an improvement in any step of microarray data analysis should be reflected in improved co-clustering of related genes. Our results suggest that background correction has little affect on GO term co-clustering characteristics, normalization has a big impact, some summarization methods constantly outperform others, some interdependencies exist among these methods, e.g., a good normalization method may work well with one summarization method but not another. An automated method for running and assessing these 300 pipelines has been made available through the Immunology Database and Analysis Portal (ImmPort; www.immport.org) - a public database resource funded by NIAID to support the management and analysis of clinical and mechanistic data generated by their funded investigators.
Use of surface modified microspheres for setting compensation in multicolor flow cytometry (144.15)
Authors: Zhang Yu-Zhong , Langsdorf Chris , Bradford Jolene 
Published: January 2023

Abstract
Yu-Zhong Zhang, Chris L. Langsdorf and Jolene A. Bradford Invitrogne, Eugene, OR 97402, USA Compensation is a process mathematically corrects multiparametric flow cytometry data for spectral overlap. Setting compensation properly requires the use of controls. Recent advances introduced antibody capture beads as substitute for cells in setting compensation. Antibody capture beads can be stained with antibodies, just like cells, and provide a bright uniform signal, regardless of how bright an antibody would stain. We compare two methods of setting compensation for multicolor immunophenotyping, using cells and using antibody capture beads, with both human lymphocytes and mouse splenocytes, and found the antibody capture beads performed comparably. It is essential that a dead cell discriminator be included in each sample, because antibody conjugates bind nonspecifically to dead cells and may lead to erroneous results. We study the need for using a dead cell discriminator in three types of assays: surface immunophenotyping, intracellular testing requiring fixation, and a functional assay. Our data indicated that excluding dead cells from that analysis improved the results. We then used the amine-reactive beads along with the antibody capture beads for setting compensation in a multicolor immunology experiment, and found both types of compensation beads used together provide a consistent, accurate, and easy-to-use technique for the setting of flow cytometry compensation.
Assay development and validation of inflammatory markers in impression cytology specimens (144.19)
Authors: Yafawi Rolla , Sace Frederick , Huang Jing-Feng , John-Baptiste Annette 
Published: January 2023

Abstract
The principle of Impression Cytology (IC) utilizes a non-invasive technique to remove superficial layers of the ocular surface. The application of a filter to the surface of the bulbar conjunctiva allows for repeated collection of superficial cells for the analysis of ocular surface disorders. Evidence suggests expression of the HLA-DR antigen on the surface of conjunctival epithelial cells is associated with allergic conjunctivitis and dry eye syndrome. Performance characteristics of HLA-DR was evaluated through intra, inter and stability assays. Evaluation of the reproducibility and overall performance of the novel methodology would be useful in understanding the tolerance level of the assay. Utilizing established techniques, IC specimens were obtained from 12 donors and assayed over multiple days and timepoints. Conjunctival epithelial cells were extracted and assayed based on differential immunostaining of both conjunctival epithelial cells and leukocytes. The reproducibility across donors over time was consistent, demonstrating conjunctival cells maintained their integrity of cell surface expression of HLA-DR. In conclusion, the studies met all validation criteria, and support the utilization of HLA-DR inflammatory measurements in impression cytology specimens. Utilization of a flow cytometric platform to examine the expression of HLA-DR antigen may help predict clinical outcome, evaluate disease progression, and monitor the effects of treatment.
Long-term functional culture of human tonsil lymphocytes in a three-dimensional bioreactor system (144.32)
Authors: Bottaro Andrea , Kuzin Igor , Sun Hongliang , Moshkani Safiekhatoon , Mantalaris Athanasios , Wu JH David 
Published: January 2023

Abstract
Peripheral lymphoid organs are the primary sites of adaptive immune responses, and harbor characteristic anatomical and histological features reflecting separation of cellular subsets (e.g. T and B lymphocytes) and functional compartments. Disruption of lymphoid organ structure by genetic mutations, pharmacological treatments or infection can lead to defective immunity, highlighting the role of tissue organization in immune responses. The generation of in vitro culture systems recapitulating salient features of peripheral lymphoid organs for experimental, biotechnological and clinical applications would be highly desirable, but has so far been hampered by the complexity of these organ systems and has encountered only limited success. We have previously developed a 3D bioreactor system capable of supporting long-term, functional human bone marrow cultures. Here we show that the same system can be adapted for culture of human primary peripheral lymphoid tissue (tonsil) cells for periods up to several weeks in the absence of specific external growth factors or activators. Cells in this system display extended survival, maintain population diversity and phenotypes largely comparable to primary cells, and organize in cellular aggregates with identifiable separation of T and B cell clusters analogous to primary organs. Strikingly, we also show that these cultures are capable of supporting the generation of antibody-producing cells in response to a panel of diverse antigens.