bluuebunny/crossref_metadata_2025_split huggingface dataset:
Oncostatin M suppress activation of IL-17/Th17 via SOCS3 regulation in CD4+ T cells of autoimmune arthritis (123.22)
Authors: Cho Mila , Lee Seon-Yeong , Son Hye-Jin , Ryu Jun-Geol , Moon Young Mee , Heo Yang Mee , Kim Ho Youn 
Published: January 2023

Abstract
Objective: We investigated whether the OSM was induced SOCS3 in Treg and Th17 generation in type II collagen induced arthritis (CIA) mice. Method: Isolated T cell of CIA mice was incubated with Th17 or Treg differentiation condition for 30min or 3day. The Th17 and Treg culture condition was incubated with or without OSM 5, 10, 20 and 50 ng/ml. The cell lysated was analyzed using western blot for phospho(p)-STAT3705, p-STAT3727 andp-STAT5 IL-17production was measured in 3day culture supernatant. IL-17 and SOCS3 mRNA levels were analysis by real-time PCR. Result: OSM was increased p-STAT3705 in Th17 and Treg culture condition, but not p-STAT5. However, production of IL-17 and IL-17 mRNA was decreased in Th17 and Treg culture condition. Inaddition, the OSM was increased SOCS3 mRNA in Th17 and Treg condition. Next we investigated expression of OSM and SOCS3 mRNA levels in Treg cell. P-STAT5, OSM and SOCS3 were activated inTreg, while p-STAT3 was decreased. Although OSM was induced activation of STAT3, that was suppressed differentiation from Treg to Th17. These result ssuggest that OSM was regulated expression of SOCS3, thereby tightly regulated plastically differentiation of Treg and Th17. Conclusion: SOCS3 induced by OSM is an acting negative feedback loop and since OSM-mediated signaling is suppression IL-17 production in mouse CD4 T cells, which is provide for feedback inhibition of inflammatory response in the CIA.
Insulin B:9-23 peptide-reactive γδ T cells in the non-obese diabetic (NOD) mouse recognize an oxidized dimeric form of the peptide (123.23)
Authors: Aydintug Mustafa , Zhang Li , Michels Aaron , Day Brian , Eisenbarth George , O'Brien Rebecca , Born Willi 
Published: January 2023

Abstract
The insulin peptide ins2 B:9-23 is recognized by diabetogenic αβ T cells in NOD mice, and by γδ T cells. Unlike the αβ T cell response to the peptide, the γδ T cell response is independent of APCs. Furthermore, the cysteine residue in position 19 of the peptide is required for the γδ but not the αβ response. Since cysteine residues can dimerize upon oxidation, we hypothesized that γδ T cells recognize oxidized dimeric peptide molecules. We therefore compared the stimulatory activities of monomeric, copper chloride-oxidized (mostly dimeric) and DMSO-oxidized, HPLC-purified dimeric insulin peptide B:9-23 preparations. We found that oxidized and purified dimeric peptide preparations stimulated γδ T cells far better than untreated or purified monomeric preparations, which still contained some oxidized peptides. In contrast, αβ T cells responded only to monomeric peptides except when live APCs were present. The γδ T cell response to the dimeric insulin peptide was TCR dependent because responsiveness could be transferred with transduced TCR-γ/δ genes derived from the originally described B:9-23-reactive γδ T cell hybridoma SP9D11. These findings provide an explanation for the differential response requirements of insulin peptide-reactive αβ and γδ T cells, and they raise the possibility that B:9-23-reactive γδ T cells function as sensors of oxidative stress.
The Role of Th17 cells in Type 1 Diabetes. (123.30)
Authors: Li Cheng-Rui , Bradley Linda 
Published: January 2023

Abstract
Type 1 diabetes (T1D) has been considered to be elicited primarily by Th1 cells. However, the role of Th17 cells, which have been implicated in other autoimmune diseases, is controversial. Using the NOD model, we found that CD4 T cells infiltrate the pancreas by 2 weeks of age, a significant proportion of which expressed either IFNγ or IL-17, suggesting that islet-specific Th1 and Th17 both develop very early. Although serum IL-17 levels increased at diabetes onset, Th17 cells from spontaneously diabetic mice failed to transfer disease to NOD.Scid recipients whereas Th17-depleted CD4 cells elicited diabetes as efficiently as non-depleted cells. To further address the pathogenic potential of Th17 cells, we differentiated polyclonal or BDC2.5 Th17 cells in vitro and transferred them into NOD or NOD.Scid recipients. Polyclonal Th17 cells failed to induce diabetes, and the recovered donor cells maintained their phenotype, suggesting that Th17 cells can be stable. In contrast, BDC2.5 Th17 cells readily induced diabetes in both recipients, although a majority of the donor cells became IFNγ-producers. However, neither the acquisition of IFNγ production nor IFNγ itself were required for diabetes induction. Instead, TNFα was essential for their pathogenicity. Our results raise the possibility that Th17 cells do not make a critical contribution to spontaneous T1D unless re-differentiation that is marked by IFNγ production occurs as a consequence of local inflammation in the pancreas.
PKC-θ shifts balance between inflammatory and regulatory T cells (123.42)
Authors: Ma Jian , Wang Ruiqing , Fang Xianfeng , Sun Zuoming 
Published: January 2023

Abstract
We demonstrate here that PKC-θ is a critical molecule to maintain immunological balance by shifting the ratio of inflammatory Th17 and inhibitory Treg cells. TGFβ-induced T regulatory cell (iTreg) differentiation was enhanced in naive PKC-θ -/- T cells or WT cells treated with a specific PKC-θ inhibitor. PKC-θ -/- T cells had reduced activity of the AKT kinase, and the expression of a constitutively active form of AKT in PKC-θ -/- T cells restored ability to inhibit iTreg differentiation. Furthermore, knockdown or overexpression of the AKT downstream targets FoxO1 and FoxO3a was found to inhibit or promote iTreg differentiation in PKC-θ -/- T cells, indicating that the AKT-FoxO1/3A pathway plays an inhibitory role in the differentiation of iTreg downstream of PKC-θ. In the other hand, we show that naive PKC-θ -/- T cells were defective in Th17 differentiation in vitro, whereas in vitro differentiation of Th1 and Th2 appeared normal. PKC-θ -/- T cells had notably lower levels of Stat3, a transcription factor required for Th17 differentiation. PKC-θ-mediated activation of the Stat3 promoter was inhibited by dominant negative AP-1 and IκB kinase β mutations, but stimulated by WT AP-1 and IκB kinase β, suggesting that PKC-θ stimulates Stat3 transcription via the AP-1 and NF-κB pathways. We conclude that PKC-θ is able to control T cell-mediated immune responses by shifting the balance between the differentiation of Th17 inflammatory and regulatory T cells.
Innate and adaptive tolerance to apoptotic cells is controlled by an IDO- dependent mechanism. (123.49)
Authors: Ravishankar Buvana , Liu Haiyun , Shinde Rahul , Chandler Phillip , McGaha Tracy 
Published: January 2023

Abstract
Marginal zone macrophages (MZMs) play a crucial role in generation of systemic tolerance to apoptotic cell (AC)-associated antigens. However, the mechanism(s) by which MZMs drive tolerogenic responses is not understood. In this report, we show that systemic administration of ACs induces splenic expression of the immunosuppressive enzyme indoleamine 2-3 dioxygenase (IDO) in Marco+SignR1+ MZMs. Moreover, we found abrogation of IDO activity altered immunity to ACs resulting in decreased TGF-β synthesis and increased pro-inflammatory cytokine production associated with a reduction in Treg function and a loss of T cell tolerance towards AC antigens. When pre-symptomatic, lupus-prone MRLlpr/lpr mice were examined we found significant IDO expression in the splenic MZ analogous to the observations in AC challenged mice. When IDO was inhibited, autoimmunity was rapidly accelerated in MRLlpr/lpr mice with a &gt;10-fold increase in anti-dsDNA IgG titers and increased renal pathology. Finally, IDO-/- mice, which do not show signs of autoimmunity, were challenged chronically with ACs. The mice developed high titer serum IgG autoreactivity, kidney inflammation, and increased mortality compared to B6 mice, which did not develop autoimmunity. Thus, the data demonstrate that a novel IDO-dependent regulatory circuit is induced in MZMs upon apoptotic cell phagocytosis providing clues to future therapeutic targets to re-establish tolerance in systemic autoimmune disease.
Green tea epigallocatechin-3-gallate modulates differentiation of naive CD4+ T cells into specific lineage effector cells (123.6)
Authors: Wang Junpeng , Pae Munkyong , Meydani Simin , Wu Dayong 
Published: January 2023

Abstract
CD4+ T cells are implicated in pathogenesis of autoimmune disease: Th1, Th9, and Th17 cells are shown to induce autoimmunity whereas regulatory T cells (Treg) may be protective. We previously showed that green tea catechin epigallocatechin-3-gallate (EGCG) attenuated experimental autoimmune encephalomyelitis (EAE) through altering CD4+ T cell subset balance, but the mechanisms are yet to be elucidated. We thereby determined how EGCG impacts differentiation of naïve CD4+ T cells into different effector lineages. Results showed that EGCG impeded Th1, Th9, and Th17 but slightly promoted Treg differentiation. In the presence of TGF-β, IL-6 promoted Th17 and suppressed Treg development; this effect of IL-6 was diminished by EGCG. Further, EGCG inhibited T-bet, PU.1, and RORγt, the transcription factor for Th1, Th9, and Th17 differentiation, respectively. EGCG also induced down-regulation of p-STAT1 & p-STAT4, and p-STAT3, the key transducers for IFN-γ-induced T-bet activation and IL-12-amplfied Th1 response, and for IL-6-induced RORγt activation, respectively. In vitro EGCG inhibited soluble IL-6Rα and membrane gp130, and plasma soluble gp130 was increased in EAE mice fed EGCG, which may contribute to reduced generation of Th17. Taken together, our results suggest that EGCG modulates development of CD4+ T cell lineages through impacting their respective regulatory networks which are also interactive, as such affording a protective effect in T cell-mediated autoimmune diseases.
Application of Brilliant Violet™ fluorescent polymer-labelled antibody reagents in 15-color flow cytometric immunophenotyping (124.12)
Authors: Lundsten Kelly , O'Connor Carolyn , Wang Elena , Li Sky , Ilic Alex , Lay Gene 
Published: January 2023

Abstract
Monitoring populations of immune cell subsets, such as B lymphocytes, T lymphocytes, NK cells, NKT cells, Treg cells, monocytes and dendritic cells, is important in many different infectious diseases, including HIV/AIDS, to assess response and recovery to treatment and therapy. In the last decade, multicolor flow cytometry has greatly increased the capabilities and consistency of immunophenotyping and immunomonitoring. However, limitations still preclude a wider adaption of &gt;10 color flow cytometry, largely due to the limited availability of fluorophores with flexible utility in a complex panel. Although instruments with 18-20 parameters are now increasingly common, until recently, the available reagents had significant practical limitations, such as limited brightness and permeability, and undesirable cross-beam excitation, resulting in reduced sensitivity when detecting lowly abundant antigens in these assaulted channels. The Brilliant Violet™ family of fluorescent probes, released in 2011, overcomes these limitations, with significantly increased overall brightness and signal-to-noise. Here, we present a 15 color/ 17 parameter flow cytometry assay, introducing Brilliant Violet™ 605 and Brilliant Violet 650™ in our expanding family of fluorescent polymers. This family of fluorescent probes will be instrumental in making a wide array of commercially available conjugates with a broad range of brightness and no limitations for intracellular detection.
A flow cytometer with a standardized fluorescence detection system simplifies multi-color panel design. (124.13)
Authors: Roys Stacey , Rogers Clare , Dinkelmann Maria 
Published: January 2023

Abstract
Multi-color panel design can be difficult due to the many factors which impact signal detection including the fluorescence detection capability of the cytometer, target molecule density and the spectral characteristics of available fluorophores (fluors). We demonstrate that a flow cytometer with a fixed and standardized fluorescence measurement system simplifies design of multi-color flow experiments, using a common conundrum researchers face as illustration: several different fluor conjugates detectable by the cytometer are available for a given clone. Which is best to use? First we show that by locking down optical alignment and optimizing fluorescence detection at the time of manufacture the BD Accuri C6 gives reproducible, predictable fluorescence measurements both for individual instruments over time and between instruments. We also show that this design leads to predictable fluorescence spillover for stable fluors. Since fluorescence spillover due to bright fluors and/or highly expressed epitopes can mask dimmer signals in neighboring channels, successfully predicting spillover greatly simplifies multi-color panel design. We illustrate this by comparing the spillover of four fluors optimally detected in FL3 of the BD Accuri™ C6 (PerCP, PerCP-Cy™5.5, PE-Cy7, BD Horizon™ PE-CF594), when conjugated to antibody clones of a high (CD45) and low (CD19) epitope-number antigen. We also quantitate the impact of the spillover on detection of weak signals in neighboring channels.
A Novel Approach to Improve Immunological Animal Study Conduct Efficiency, Data Integration, Consistency and Security and the Sharing of Results (124.16)
Authors: Kumer Jeff , Ibsen Eric 
Published: January 2023

Abstract
The traditional management of immunological animal studies and results is challenging due to complex, varied workflows as well as the acquisition, analysis, secure storage and accessibility of disparate data types acquired and stored in multiple formats and locations. Existing approaches often poorly reflect the details of planned-versus-actual study conduct, animal welfare considerations, and integration of data types in various paper and electronic formats. Errors and non-standardized condition scoring, data acquisition, processing, analyses are common problems as is impeded access to current and historical results due to compartmentalized data silo storage. The result is a highly inefficient use of financial, labor and animal resources and the unintentional loss of historical results over time. In-house-developed and commercially-available desktop and web-based applications have been designed specifically to manage complex and variable study designs, comprehensive task workflow and information management. For the first time, disparate data types from immunology studies can be acquired consistently, linked, analyzed, graphed and securely shared with stakeholders in real time and in perpetuity saving precious research resources and minimizing historical information loss. Several non-commercial and commercially-available solutions will be described and their respective impacts on immunology study workflow efficiency and data quality, integration, security and accessibility.
A specialized tube to make enrichment of specific cell subsets faster and easier (124.3)
Authors: Fairhurst Maureen , Fadum Jodie , Woodside Steve , McQueen Karina , Thomas Terry , Peters Carrie 
Published: January 2023

Abstract
Many experimental protocols require the enrichment of specific cell subsets from peripheral blood. RosetteSep™ cell enrichment and standard mononuclear cell (MNC) preparation both involve density gradient centrifugation, which entails slowly layering the sample over the density gradient medium to avoid mixing, and carefully pipetting to remove the enriched cells after centrifugation. Centrifugation must be performed with the brake off to avoid disturbing the enriched cell layer, further lengthening the process. SepMate™, a centrifugation tube with a specialized insert, was developed to allow rapid layering of the sample onto the density gradient medium, and pouring off of the enriched cells after centrifugation, thus simplifying the entire process. When using SepMate™, the cocktail incubation time and centrifugation time could each be shortened to 10 min, making RosetteSep™ cell enrichment even faster. RosetteSep™ enrichments of mononuclear cell subsets using the SepMate™ tubes and protocol gave equivalent purity and recovery of desired cells compared to using the standard RosetteSep™ protocol, and desired cells could be enriched from whole blood in &lt;30 min. Purities of specific cell types were: CD3 T Cells 96 ± 1 (n=5), CD4 T Cells 94 ± 5 (n=3), CD8 T Cells 85 ± 11 (n=4), B Cells 92 ± 6 (n=3), NK Cells 85 ± 5 (n=5), monocytes 68 ± 8 (n = 4). The protocol is easily scalable to process multiple samples simultaneously, and the SepMate™ tube can also be used to prepare MNCs.
Multiplex measurement of inflammatory cytokines in human plasma using a Fluorokine® MAP multiplex assay (124.5)
Authors: Schmidt Jane , Johnson Paige , Anderson Michael 
Published: January 2023

Abstract
Fluorokine® Multianalyte Profiling (MAP) Kits offer the ability to simultaneously measure multiple analytes with less operator time, less sample consumption, and lower cost per result compared to conventional ELISAs. To demonstrate this, we cultured whole blood samples from apparently healthy donors for up to 24 hours without stimulation or stimulated by LPS, PHA, or PMA. Samples were taken from the cultures and processed into plasma. GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, TNF-α, and VEGF were quantified simultaneously using the Human Fluorokine MAP High Sensitivity Cytokine Panel to compare cytokine profiles in stimulated and unstimulated cultures. The Human Fluorokine MAP High Sensitivity Cytokine Panel is a fully validated multiplex assay designed to measure up to twelve inflammation biomarkers in serum or plasma; the specific analytes are selected by the user. Analyte-specific antibodies pre-coated onto color-coded microparticles capture the analytes of interest in standards or samples. After washing away any unbound substances in a filter-bottom microplate, a cocktail of biotinylated antibodies also specific to the analytes of interest is added to each well, followed by a streptavidin-phycoerythrin conjugate. Following a final wash, microparticles are resuspended in buffer and read using a Luminex®-based analyzer. The assay is fully validated for accurate, precise, and reproducible results using serum or plasma samples.
Optimiser™ Microplate enables simultaneous multi-analyte quantitation using ultra-low sample volumes in a 2-hour assay: Validation of a Novel 10-Analyte Th17 Cell Panel (124.7)
Authors: Kai Junhai , Moore Victor , Puntambekar Aniruddha , Sehy David , Lakkis Montaha , Lee Sean , Han Jungyoup , Ahn Chong 
Published: January 2023

Abstract
Optimiser™ Microfluidic ELISA Plates combine microfluidics with 96-well architecture to offer dramatic improvements over conventional ELISA, such as 50fold-reduced sample volume, 2fold-reduced assay time, and ultra-high sensitivity, while still using conventional ELISA reagents. Optimiser™ ELISA Plates are compatible with existing pipetting systems (manual or automated) and ELISA plate readers. Furthermore, the surface area to volume ratio of each microfluidic reaction chamber represents a 50-fold increase when compared to standard plate microwells. The improvement in surface:volume ratio increases binding kinetics dramatically, enabling assays to be run in less than half the time of traditional ELISAs, and providing comparable sensitivity with as little as 2 microliter samples (i.e., 50fold less than required for conventional assay). A multi-analyte assay for detecting products of activated Th17 cells (e.g., IL-6, IL17A, IL-17AF, IL-17FF, IL-21, IL-22, IL-26, G-CSF, CCL20, TNF-α) is used to demonstrate that using only 25 microliter sample volume, the Optimiser™ ELISA can simultaneously measure 10 analytes (with sensitivity comparable to conventional ELISA) in a 2-hour assay, without the need for dedicated multiplexing instruments, multiplexing reagents, or bead-labeling chemistry. The entire 10-analyte assay procedure is fully automated and is run on a Tecan EVO100™ System.
A novel mechanism responsible for IgE regulation involves beta2-adrenergic receptor-mediated regulation of CD23 and ADAM10 localization to B cell-derived exosomes. (125.11)
Authors: Padro Caroline , Sanders Virginia 
Published: January 2023

Abstract
Allergic asthma severity positively correlates with plasma IgE, but physiological mechanisms responsible for regulating IgE production by B cells are unknown. We have shown that beta-2 adrenergic receptor (β2AR) engagement on a CD40L/IL-4-primed B cell increases the level of IgE produced per cell, without affecting class switch recombination. Asthma therapy involves stimulation of the β2AR on bronchiolar smooth muscle cells to relieve bronchoconstriction. This therapy also targets the β2AR expressed on primed B cells to increase IgE production, potentially intensifying symptoms. In this study we show that surface CD23 remained constant on primed B cells exposed to a β2AR agonist, while CD23 expression and soluble CD23 (sCD23), which is a positive regulator of IgE production, increased. We hypothesized that the primary sheddase of CD23, ADAM10, is regulated by β2AR engagement on a primed B cell. We found that CD23 and ADAM10 are expressed on B cell-derived exosomes over time and that blockade of exosome release allows for CD23 and ADAM10 retention, with β2AR engagement augmenting CD23 and ADAM10 retention. Localization of ADAM10 and CD23 to non-lipid raft fractions where CD23 is cleaved by ADAM10 was also increased by β2AR engagement. Thus, ADAM10 may serve as a molecular target that can be blocked during β2AR drug therapy to prevent increased sCD23 and IgE production, while maintaining the therapeutic effect of bronchorelaxation.
Immunotherapy reduces IL-33 and IL-6 expression in the lung of T-bet deficient mice in an experimental model of allergic asthma (125.9)
Authors: Mousset Stephanie , Koch Sonja , Finotto Susetta 
Published: January 2023

Abstract
Immunotherapy (IT) is one of the most effective therapies for increasing allergic diseases worldwide including allergic asthma. Previously, we have established a murine model of subcutaneous IT for experimental allergic asthma. We demonstrated that main components of allergic asthma induced by T-bet deficiency such as airway hyperresponsiveness could be ameliorated. These data suggested that the molecular mechanism induced by IT is T-bet independent. To analyze this yet unknown mechanism we started to investigate IL-6 expression in this model. IL-6 is a cytokine produced by several cell types of the innate as well as adaptive immune system. We previously described that IL-6 is increased in the lung cells of T-bet deficient (T-bet-/-) naïve mice as compared to wild type littermates. In a murine model of allergic asthma we found that IL-6 is significantly down-regulated in the lung of T-bet-/- mice that have been treated according to the IT protocol. Likewise, we found that the newly discovered cytokine IL-33, of the IL-1 family, is significantly up-regulated in the lung of T-bet-/- mice in experimental asthma and down-regulated after IT. In conclusion we discovered that the protective mechanism of the IT comprises down-regulation of IL-6 and IL-33. These findings could be considered for future novel therapy for allergic asthma.