bluuebunny/crossref_metadata_2025_split huggingface dataset:
Oxidation resistance protein 1 (Oxr1) protects against autoantibody-induced glomerulonephritis by inhibiting inflammation and apoptosis (56.10)
Authors: Li Yajuan , Shao Xinli , Raman Indu , Yan Mei , Du Yong , Mohan Chandra , Li Quanzhen 
Published: January 2023

Abstract
Oxidative stress is a common pathologic feature of immune-mediated nephritis in both human and mouse model. However, the molecular pathways that govern defense against oxidative stress-induced renal injury remain unclear. Using experimental autoantibody-induced nephritis mouse model, we have identified OXR1 may play a protective role against anti-GBM disease and lupus nephritis. Impaired expression of OXR1 was observed in the kidneys of 129/svj and DBA1 mice that suffered from severe anti-GBM-antibody induced nephritis, compared with non-disease controls. The reduced expression of OXR1 was also detected in kidneys of congenic mice with spontaneous lupus nephritis (e.g., B6.Sle1.Sle3 and B6.Mrl/Lpr). Deliberate delivery of human OXR1 via lentiviral vector into mouse kidney attenuated anti-GBM-antibody induced nephritis in 129/svj mice. The overexpression of hOXR1 in recipient mice down-regulated the expression of several inflammatory chemokines, including RANTES, SDF-1, IL1β and IL6, and suppressed the infiltration of macrophage/lymphocyte in the kidneys. The increased OXR1 was also associated with reduced renal cell apoptosis in anti-GBM mice. In vitro experiment further proved that hOXR1-transfected macrophages are more resistant to H2O2-induced apoptosis. These findings suggest that OXR1 may play a protective role in immune-mediated nephritis by modulating inflammation and oxidative stress and therefore may be a potential therapeutic target for lupus nephritis.
Follicular B cells, not Marginal zone B cells, preferentially contribute to development of spontaneous autoimmune thyroiditis (SAT) and are targets of anti-CD20 therapy (56.12)
Authors: Hong So-Hee , Mullen Helen 
Published: January 2023

Abstract
SAT in NOD.H-2h4 mice is a chronic organ-specific autoimmune disease characterized by mononuclear cell infiltration of the thyroid and destruction of thyroid follicles by infiltrating inflammatory cells. Like other autoimmune diseases, B cells play a critical role in pathogenesis of SAT as important antigen presenting cells (APCs) for activation of CD4+ T cells. However it is not yet known which subsets of B cells are important for SAT development. In this study, we found that expression of the co-stimulatory molecules CD80 and CD86 was significantly increased on splenic CD23hiCD21low follicular (FO) B cells, but not marginal zone (MZ) B cells, after SAT development, but absolute numbers and percentages of splenic FO B cells were not altered. In draining lymph nodes, CD40 expression on FO B cells was also increased in mice with SAT. All thyroid infiltrating B cells in mice with SAT were FO B cells. Depletion of FO B cells by anti-CD20 antibody ameliorated established SAT, and anti-thyroglobulin antibody levels were reduced. Moreover, administering anti-CD20 to mice with established SAT results in depletion of B cells in the thyroid and depletion of thyroid B cells is longer lasting than depletion of splenic B cells. On the basis of these findings, we suggest the FO subset of B cells preferentially contributes to SAT development and persistence, and targeting FO B cells after disease develops provides an effective means to treat autoimmune diseases.
Rapid, column-free two-step procedure for the enrichment of human Th17 Cells from peripheral blood (56.13)
Authors: Fairhurst Maureen , Yu Jessie , Thomas Terry 
Published: January 2023

Abstract
Human Th17 cells play a critical role in the regulation of autoimmune disease and antimicrobial host defenses through production of pro-inflammatory cytokines IL17A and IL-17F. They can be characterized by expression of surface receptors including IL-23R, CCR6, CD161 and lineage-specific transcription factor RORC, but lack of CXCR3 expression. The major disadvantages of current isolation methods are the requirement for in vitro stimulation and the co-secretion of IFN-γ. We have developed a 2-step EasySepTM immunomagnetic column-free method for the enrichment of CD4+CXCR3-CCR6+ cells from fresh peripheral blood nucleated cells (PBNC). First, non-CD4 T cells and CXCR3+ cells are targeted for depletion using dextran-coated magnetic particles and a cocktail of antibody complexes. Labeled cells are separated using an EasySepTM magnet, and pre-enriched CD4 T-cells are poured off. Next, CCR6+ cells are positively selected from the pre-enriched fraction. The procedure can be automated using RoboSepTM. Starting with a frequency of 5 ± 2% CD4+CXCR3-CCR6+ cells, purities of 94 ± 3% (n=10) can be obtained. The resulting cells produce high levels of IL-17 with minimal IFN-γ when measured by ELISA and intracellular staining and increased RORC mRNA expression over total CD4 T cells and CD4+CXCR3+ T cells (Th1 cells). Enrichment of unstimulated human Th17 cells enables the investigation of adaptive immune responses and regulation mechanisms required for the development of future therapies.
Statins and rapamycin synergistically induce dendritic cell KLF2 expression and tolerogenic function in vivo (P1610) (56.8)
Authors: Bu De-xiu , Griffin Gabriel , Grabie Nir , Lederer James , Lichtman Andrew 
Published: January 2023

Abstract
The statins, a class of HMG CoA-reductase inhibitors display immunomodulatory properties, independent of their cholesterol-lowering effects. We recently demonstrated that statins modulate T cell function through upregulation of the transcription factor, kruppel-like factor 2 (KLF2). Dendritic cells are essential for initiating and regulating T cell responses. We therefore examined how statins might modulate DC function. We found that: 1) statins upregulate Klf2 gene expression in both splenic and bone marrow derived mouse DCs; 2) the combination of the mTORC1 inhibitor rapamycin and statin synergistically enhance DC Klf2 and chemokine receptor CCR7 expression at doses of rapamycin alone that have no effect on Klf2; 3) statin- plus rapamycin-treated DCs or DCs expressing Klf2 encoded by a retroviral vector skew T cell differentiation away from inflammatory pro-atherogenic Th1 and Th17 phenotypes and promote Th2 and regulatory T cell differentiation in vitro; 4) statins directly block mTOR activation in DCs as indicated by assays of phosphorylation of mTOR and its substrates; and 5) statin plus rapamycin treatment impaired the ability of myosin-peptide loaded DCs to induced experimental autoimmune myocarditis (EAM) when transferred to BALB/c mice. We conclude that pharmacologic targeting of KLF2 in T cells and DCs is a feasible goal for suppressing immune response in human disease.
Thymic stromal lymphopoietin is produced by dendritic cells. (57.3)
Authors: Kashyap Mohit , Yrina Rochman , Spolski Rosanne , Samsel Leigh , Leonard Warren 
Published: January 2023

Abstract
Thymic stromal lymphopoietin (TSLP) is a type 1 cytokine that contributes to lymphopoiesis and the development of asthma and atopic dermatitis. TSLP acts on multiple lineages, including dendritic cells (DCs), T cells, NKT cells, eosinophils, and mast cells, mediating proliferation and survival and linking innate and adaptive immune responses. TSLP is produced by a range of cells, including epithelial cells, fibroblasts, stromal cells, and keratinocytes. DCs are important primary targets of TSLP, and we unexpectedly demonstrated that DCs also produce TSLP in response to TLR stimulation and that this is augmented by IL-4. Moreover, we demonstrated that when mice were challenged with house dust mite extract, lung CD11c(+) DCs expressed TSLP mRNA at an even higher level than did epithelial cells. These data suggested that DCs not only respond to TSLP but also are a source of TSLP during pathogen and/or allergen encounter.
Adrenergic regulation of p38 MAPK phosphorylation by hematopoietic protein tyrosine phosphatase in human B cells (57.4)
Authors: McMaken Sara , Gormley Matthew , Sanders Virginia 
Published: January 2023

Abstract
Asthma affects ~300 million people and is responsible for $14 billion in annual medical costs. Therapy utilizes agonists to stimulate the beta-2-adrenergic receptor (β2AR) on lung smooth muscle cells. However, β2AR stimulation on a B cell responding to an allergen in vitro or in vivo increases IgE levels without affecting isotype switching. Elucidating how the β2AR agonist increases IgE production is crucial in blocking this effect on the B cell while maintaining bronchodilation. We have shown that β2AR stimulation on B cells during CD40L/IL-4 priming in vitro induces a PKA-dependent phosphorylation of hematopoietic protein tyrosine phosphatase (HePTP). This releases bound p38 MAPK and allows for its phosphorylation. β2AR deficiency or depletion of the endogenous β2AR agonist norepinephrine in vivo decreases levels of IgE made per B cell and lung inflammation after antigen exposure. It is currently unknown if human HePTP functions in this capacity. We sought to determine if HePTP in human B cells contributed to the β2AR-mediated phosphorylation of p38 MAPK. In comparison to anti-CD40/IL-4-primed CL01 B cells, primed B cells exposed to β2AR agonist had increased HePTP phosphorylation and p38 MAPK phosphorylation. This finding suggests that stimulation of the β2AR on human B cells elevates p38 MAPK phosphorylation through a change in HePTP function to elevate IgE. Thus, HePTP may represent a novel target for regulating IgE levels in human B cells during beta-agonist therapy.
Evaluation of the effect of pollen lipid on iNKT cells from pollen-sensitized patients (57.6)
Authors: Fereidouni Mohammad , Jabbari Azad Farahzah , Faridhosseini Reza , Varateh Abdolreza 
Published: January 2023

Abstract
Introduction: iNKT cells are a small subset of T lymphocytes which express a invariant TCR and recognize lipid antigens. They are implicated in a range of immune responses but their role in allergy is controversial. Pollens are the most important trigger of allergic symptoms but their lipid contents may also have a role in allergy through interaction with iNKT cells. The aim of this study was to evaluate the in vitro effect of pollen lipid extract on iNKT cells. Subjects and methods: PBMCs from 18 pollen-sensitized subjects and 11 healthy controls were isolated and stimulated for 7 days with a mix crude lipid extract of five allergenic pollens prepared by Folch method. Alpha-galactosylceramide and medium were used as positive and negative controls respectively. Proportion of iNKT cells was measured by FACS using 6B11 and anti-CD3 monoclonal antibodies. Results: At day 0, the mean percentage of iNKT cells was 0.42 ± 0.3% and 0.44 ± 0.2% (P &gt; 0.05) for sensitized and control groups respectively. After 7 days culture, iNKT cells from both groups expanded efficiently in response to alpha-galactosylceramide and iNKT proportion reaches to 8 ± 5.6% but lipid extract did not stimulate iNKT proliferation in any groups and the proportion of iNKT cells remained unchanged. Conclusion: The results of this study suggest that pollen lipids are not potent stimulator of iNKT proliferation but further studies need to evaluate the presence and frequency of pollen-specific iNKT cells.
Metagene projection strategies for discovering mechanisms of T cell differentiation in gene expression profiling data (58.13)
Authors: Alexe Gabriela , Tamayo Pablo , Mesirov Jill , Gupta Prakash , Wolski David , Lauer Georg , Haining William 
Published: January 2023

Abstract
Characterizing the phenotypic heterogeneity in memory T cells generated following antigen encounter is a major challenge in immunology. Tools such as CyTOF flow cytometry or gene expression profiling of T cells can capture increasingly large numbers of parameters. However analysis of high dimensional data with standard techniques such as principal component analysis or hierarchical clustering often fails to capture the biological basis for observed phenotypic heterogeneity. We address this problem by applying a metagene projection strategy based on independent component analysis and consensus non-negative matrix factorization clustering. This approach assumes that the gene expression profile of a population of CD8+ T cells is generated by groups of genes that are representative of different biological processes, and deconvolves the data into individual components (or metagenes) that together comprise some or all of the overall “signal” in the data-space. The metagene projection strategy is able to provide a robust low dimensional description of transcriptional (or other complex) data based on groups of genes that represent known and novel biological mechanisms. It is also able to provide sensitive clustering and classification solutions. We illustrate the ability of this metagene projection strategy to capture novel molecular patterns in T cell differentiation states common to antigen-specific CD8+ T cells responses to HIV, HCV and metastatic melanoma.
Dissolvable microneedle arrays deliver live adenovirus to the skin for genetic immunization. (58.16)
Authors: Erdos Geza , Donahue Cara , Zhang Jiying , Ozdoganlar Burak , Gambotto Andrea , Falo Louis 
Published: January 2023

Abstract
Recombinant DNA vaccines show incredible promise for the prevention of human disease by their capacity to effectively inducing both humoral and cellular immune responses. Among the available technologies adenovirus (Ad) vectored vaccines are among the most appealing. To date the clinical deployment of Ad-based vaccines has been limited by several considerations including the lack of delivery technology to enable safe, reproducible and effective vector delivery to patients. The skin is readily accessible and rich in antigen presenting cells, making it a preferred target site for immunization. To enable Ad-based clinical immunization, we developed microneedle arrays (MNAs) to deliver live Ad-vectors to the skin. The MNA design and fabrication process we developed results in MNAs that effectively preserve Ad viability, and enable efficient delivery of infectious vector to the skin. Specifically, MNA embedded live Ad5 virus retained infectivity for up to a year. Topical application of MNAs resulted in transduction of skin cells. Further, immunization of mice with an Ad5 expression vector encoding the SIVmac239 gag antigen resulted in the generation of potent CTL activity against the dominant gag 76-84 eptiope. Taken together, our results demonstrate the capacity of MNAs to preserve and deliver Ad5 virus to the skin for genetic immunization. This novel technology platform may enable a wide range of topical skin-targeted Ad vaccines.
Fusion of TDG and RHD for selective DNA demethylation and transcriptional enhancement of NOS2 (58.2)
Authors: Fedulov Alexey , Gregory David , Mikhaylova Lyudmila 
Published: January 2023

Abstract
DNA methylation is a key mechanism in epigenetic silencing of transcription. Removal of methylation marks in promoters and other key regulatory sequences is associated with increased gene transcription, which could be beneficial for immunomodulatory and other purposes. However there is no method to selectively demethylate DNA in living cells. We designed a molecular construct in which a putative demethylase thymine-DNA-glycosylase (TDG) is targeted to a specific sequence by fusion with a DNA binding domain of NF-κB, the Rel-homology domain (RHD). 3T3 murine fibroblasts were transduced with fusion TDG-RHD or control constructs. In these cells the promoter of inducible nitric oxide synthase (NOS2) gene is highly methylated and contains RHD-binding sequences. We observed decreased DNA methylation (by 5-10 %, p&lt;0.05) in several C’s in the NOS2 promoter and nearest CpG island. We also observed decreased methylation in the vicinity of RHD-binding sites in LINE-1, suggesting that the effect was available across the genome. Stimulation with IFNγ (250 ug/ml, 6 hrs) led to increased NOS2 message (fold over unstimulated 97.7±27.3 vs. 18.9±6.3 in control, p&lt;0.01) and protein, indicating increased gene function. The effect was not seen for control genes lacking either RHD-binding sites or high levels of methylation, nor in mock-transduced cells. Results suggest that direct targeting of TDG to specific DNA loci may be practical for selective re-activation of epigenetically silenced genes.
Quantitative phosphoproteomic analysis of Y3F SLP-76 mutant in T cell signaling pathway reveals new feedback pathways (58.20)
Authors: Ji Qinqin , Ding Yiyuan , Salomon Arthur 
Published: January 2023

Abstract
SRC homology 2 (SH2) domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) is a cytosolic adaptor protein that plays an important role in the TCR mediated T-cell signaling pathway. SLP76 consists of three functional domains: an amino-terminal acidic domain with three TCR inducible tyrosine phosphorylation sites; a central proline-rich domain and a carboxyl-terminal SH2 domain. Previous studies indicated all three of these domains contribute to the function of SLP-76. In our work, phosphoproteomic methods in mass spectrometry were used to perform a quantitative phosphoproteomic analysis comparing SLP-76 reconstituted wild-type cells with cells in which Tyr113, Tyr128, and Tyr145 were mutated to phenylalanine (denoted as the Y3F mutant). The abundance of thousands of unique phosphorylation sites was determined in the presence of wild type or Y3F mutant SLP76 through a time course of receptor stimulation. Our data provides a highly detailed new glimpse into the network of phosphorylation sites regulated through tyrosine phosphorylation of SLP76 including feedback regulation of T cell receptor proximal signaling.
The role of peptide-MHC ligand density in stimulating T-cell receptor signaling (58.21)
Authors: Sykulev Yuri , Anikeeva Nadia , Gakamsky Dimitry 
Published: January 2023

Abstract
Cell surface MHC proteins form clusters on the cell membrane. The MHC density within clusters could affect recognition of noncognate (self) and cognate peptide-MHC (pMHC) ligands and influence the kinetics of TCR-mediate signaling and quality of T cell responses. To evaluate the role of density, orientation and valency of pMHC ligands we utilized different scaffolds to assemble pMHC oligomers modeling MHC clusters. We systematically investigated how these parameters influence the binding of the pMHC oligomers to live CD8+ cytotoxic T lymphocytes (CTL) and the kinetics of TCR-mediated signaling. We have found that the recognition of noncognate or self pMHC strongly depends on the pMHC density and architecture and size of the scaffolds. In contrast, the valency but not the pMHC density mostly determined the recognition of strong agonist pMHC ligands. In addition, the pMHC density significantly influences the ability of cognate and noncognate pMHC to cooperate and to induce robust and rapid TCR signaling. The latter regulates the kinetics of target cell destruction by CTL that is essential for successful anti-virus immunity.
Protein microarray analysis of serum factor-reactive autoantibodies reveals BAFF-binding autoantibodies in systemic lupus erythematosus (58.22)
Authors: Price Jordan , Kemmer Dodge , Balboni Imelda , Haddon David , Mandelbaum Gil , Delepine Guillaume , Sokolove Jeremy , Shum Anthony , Browne Sarah , Utz Paul 
Published: January 2023

Abstract
Autoantibodies to cytokines and chemokines have been shown to influence immune pathways in patients afflicted with a range of immune pathology. We hypothesized that a multiplex platform capable of detecting antibodies to cytokines, chemokines and other immune factors would allow us to identify novel biomarkers of immune diseases and uncover new mechanisms of disease pathogenesis. We created a planar microarray containing a library of recombinant human proteins deposited on nitrocellulose-coated slides. We resolved binding of commercially available antibodies to cytokines and chemokines, and detected previously described cytokine reactivity in samples from patients with non-tuberculosis mycobacterium infection and autoimmune polyendocrine syndrome type 1. We further used the arrays to profile samples from three cohorts of systemic lupus erythematosus (SLE) patients. Among several cytokine targets, the significance analysis of microarrays algorithm identified that B-cell activating factor (BAFF)-binding autoantibodies correlated with subgroups of SLE patients. Reactive sera inhibited the activity of BAFF in a reporter cell assay, indicating that BAFF autoantibodies may influence the activity of this critical cytokine in SLE. Our results demonstrate that serum factor protein microarrays can be used to detect novel and potentially relevant autoreactivity to cytokines in human patient samples, and that BAFF-reactive autoantibodies may play a role in the pathophysiology of SLE.
Cooperation of dendritic cells with naïve lymphocyte populations to induce the generation of antigen-specific antibodies in vitro (58.3)
Authors: Heilmann Katja , Wand Inga , Holzlöhner Pamela , Micheel Burkhard 
Published: January 2023

Abstract
The production of monoclonal antibodies by hybridoma technology is dependent on lymphocytes taken from vertebrates which have to be immunized against the corresponding antigen. We present here our first experiments which should allow the replacement of this in vivo immunization step by an in vitro immunization procedure. This work provides new possibilities for the specific activation of immune cells in order to use them for the generation of antibodies which are not of murine origin. Bone marrow-derived dendritic cells were loaded with antigen and co-cultured with naïve T and B lymphocytes of non-immunized mice. The interaction and activation of the different cell types were investigated by measuring the expression of specific cell surface markers, the release of activation-dependent interleukins and the secretion of antigen-specific antibodies. We could demonstrate that dendritic cells process and present antigen fragments and activate T cells, that T cells proliferate and release activation-induced interleukins, and that B cells maturate under the influence of activated T cells and secrete antigen-specific antibodies.
Quantifying selection in high-throughput Immunoglobulin sequencing datasets (58.4)
Authors: Yaari Gur , Uduman Mohamed , Kleinstein Steven 
Published: January 2023

Abstract
Quantifying antigen driven selection based on mutation patterns observed in Immunoglobulin (Ig) DNA sequences can provide insights into the basic biology that underlies the adaptive immune response, and may further serve as diagnostic and prognostic markers. High-throughput sequencing approaches make large-scale characterization of B cell Ig repertoires feasible. However, analyzing selection in these large datasets, presents fundamental challenges. Here I will present a new computational framework for Bayesian estimation of Antigen-driven SELectIoN (BASELINe) in Ig sequecnes. BASELINe provides a more intuitive means to analyze selection by shifting the problem from one of detecting selection to one of quantifying it. It allows, for the first time, comparative analysis between sequences derived from different germlines. We have made this method practical for complete repertoire analysis through an online tool (clip.med.yale.edu/baseline) and a highly optimized implementation that can analyze up to 10,000 sequences in under four minutes on a single processor. By testing this method on computer simulations as well as on real sequences from genetically engineered mice, we have shown that indeed there is a significant improvement in our ability to quantify selection strength. We then demonstrate how BASELINe can be applied to high throughput sequencing datasets to arrive at several novel non-trivial observations.