bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD39-mediated Effect of human mesenchymal stem cells on the human Th17 cells (P1221)
Authors: Kim Mee Kum , Jeong Hyun Jeong , Kim Dong-Hyun , Wee Won Ryang , Lee Won Woo 
Published: January 2023

Abstract
Mesenchymal stem cells (MSCs) have been known to have anti-inflammatory properties in various inflammatory diseases. However, the reported effects of human MSCs on human Th17 cells have not been confirmed. This study was aimed to investigate inhibitory effects of human mesenchymal stem cells (hMSCs) on human CD4+ T cells through CD39 mediated pathway. The study was performed in accordance with the guidelines of the Declaration of Helsinki and was approved from the Institutional Review Board of the Seoul National University Hospital. Human peripheral blood mononuclear cells was obtained from the healthy voluntary donors with informed consents(n=6). Bone marrow-derived hMSCs were co-cultured with human CD4+ T cells at a MSC:T cell ratio of 1:10. The suppressive effects of MSCs were evaluated by assessing their effects on the proliferation of CD4+ T cells by carboxyfluorescein diacetate succinimidyl ester (CFSE) staining and the secretion of IFN-γ and IL-17A by CD 4+ memory T cells with or without anti-CD39 treatment. hMSCs effectively suppressed the proliferation and both IFN-γ and IL-17A secretion of CD4+ CD45ROhi T cells, while, anti-CD39 treatment significantly reduced the inhibitory function of hMSCs on the CD4 T cells. The expression of CD73 on the T cells was highly induced with co-culture of hMSCs. It suggests that hMSCs effectively suppressed the function of CD4+ T cells which was mediated by the CD39 and CD73.
Mechanism of combinatorial cytokine effects on dendritic cell state (P1200)
Authors: Hartmann Boris , Zaslavsky Elena , Marjanovic Nada , Moran Thomas , Sealfon Stuart 
Published: January 2023

Abstract
The effects of cytokine microenvironment on cellular response state are potentially important and relatively unexplored in the immune system. We focus on elucidating the combinatorial effects of extracellular signaling in the dendritic cells (Dc) and the mechanisms responsible for decoding these signals. Newcastle disease virus infected human dendritic cells in culture secret 21 extracellular factors for which they express receptors. Previously we showed that paracrine cytokines involving more than type I interferon released by virus-infected human Dcs dictate an antiviral maturation state of naïve Dcs. Microarray studies identified 260 genes that are differentially regulated (p≤0.05; &gt;2-fold) by infected Dc supernatant compared to cells treated with IFNβ alone. Combinatorial cytokine studies indicate that IFNβ and TNFα are major factors that together are responsible for stimulating this state in Dcs. Linear regression modeling of microarray data identified 727 synergistically regulated genes and gene set enrichment analysis implicated STAT-5, STAT-6 and IRF-1. The combinatorial activation of these transcription factors was confirmed by imaging flow cytometry. These results suggest that the cytokine microenvironment generate Dc response states via the activation of combinatorial-responsive transcription factors. Combinatorial regulation mechanisms may provide the ability for localized control of the activation state of immune cells to spatially tune the response to infection.
Human metapneumovirus infection induces expression of thymic stromal lymphopoetin and IL-33 in airway epithelial cells in vitro and lung infiltration of OX40L+ DCs in vivo (P6200)
Authors: Lay Margarita , Cespedes Pablo , Palavecino Christian , Leon Miguel , Bueno Susan , Kalergis Alexis 
Published: January 2023

Abstract
Human metapneumovirus (hMPV) is a common virus known to induce acute respiratory tract infection. The specific mechanism by which hMPV modulates the immune response in infected airway epithelial cells (AECs) leading to tissue damage, still remains unknown. Thymic stromal lymphopoetin (TSLP) and IL-33, epithelial cell derived cytokines, induce high expression levels of OX40L on DCs, leading to Th2 responses and allergic inflammation. Recent reports have shown that Th2 inflammatory responses in RSV infections are mediated by TSLP-dependent OX40L expression in DCs. We hypothesize that the TSLP, IL-33 and OX40L pathway mediates lung inflammation in hMPV-infected individuals. We tested the expression of TSLP and IL-33 in human AECs after hMPV infection and compared them with RSV-infected AECs by qRT-PCR. We also evaluated lung infiltration of GR1+CD11b+ and CD11c+CD11b+OX40L+ cells in hMPV-infected mice by flow cytometry. We found an induction of TSLP mRNA levels in hMPV-infected AECs in a MOI dependent-manner. We also proved that hMPV induces higher TSLP mRNA levels than RSV in AECs, and hMPV but not RSV up-regulates IL-33 levels in AECs. Furthermore, we showed that hMPV infection in mice induces an increased lung infiltration of neutrophiles and OX40L+ DCs than the mock control group. These results suggest that hMPV infection in lungs may be inducing an augmented Th2 inflammatory response, through TSLP and IL-33 produced by hMPV-infected AECs and activated DCs expressing OX40L.
Narrowing of influenza A virus specific T-cell receptor alpha and beta repertoire with increasing age (P6176)
Authors: Gil Anna , Yassai Maryam , Naumov Yuri , Selin Liisa 
Published: January 2023

Abstract
Alterations in memory CD8 T cell responses may contribute to the high morbidity and mortality caused by seasonal influenza A virus (IAV) infections in older individuals. We questioned whether memory CD8 responses changed overtime with increasing age to this non-persistent virus, to which recurrent exposure with new strains is common. Here, for the first time we show that with age the HLA-A2-restricted IAV M158-66-specific Vα T cell repertoire was significantly narrowed leading to oligoclonal expansions including the usage of a single identical VA12 clonotype in all 8 older donors. The VA repertoire of older individuals also had longer CDR3 regions with increased usage of alanine/glycine runs which may enhance TcR promiscuity. Collectively these results suggest that CD8 memory responses in humans to non-persistent viruses like IAV are dynamic, and with aging become narrower with preferential retention of T cell repertoires with features of enhanced promiscuity.
B cell activation is regulated by the stiffness properties of the substrate presenting the antigens (P1152)
Authors: Liu Wanli , Wan ZhengPeng 
Published: January 2023

Abstract
B lymphocytes are activated upon the sensing of antigens by BCRs. The substrate presenting the antigens can show different degree of stiffness. However it is not clear if B cells can respond to substrate stiffness changes. In this report, we use high resolution, high speed live cell imaging techniques to capture the early events in B cell activation shortly after the recognition of antigens tethered to polyacrylamide gel substrates with variable degrees of stiffness as quantified by Young’s modulus (2.6-22.1 kPa). We show that the initiation of B cell activation is extremely sensitive to substrate stiffness. B cells exhibit much stronger activation responses when encountering antigens tethered to substrates with a high degree of stiffness as measured by the accumulation of BCRs, pSyk and pTyr molecules into B cell immunological synapse. Moreover, BCR and pSyk microclusters induced by antigens on stiffer substrate are more prominent and show significantly enhanced colocalization compared to activation with antigens on softer substrate. As a result, the expression of the B cell activation marker CD69 is greatly enhanced in B cells encountering antigens on stiffer substrate. Using a series of cytoskeleton inhibitors, we determine that the mechanosensing capability of B cell is strictly dependent on microtubules, and only mildly linked to actin cytoskeleton. These results suggest the importance of mechanical properties mediated by substrate stiffness in B cell activation.
TRAF6 regulates TCR signaling via interaction with and modification of LAT adapter (P1178)
Authors: Yingqiu Li , Xie Ji-Ji , Liang Jia-Qi , Altman Amnon 
Published: January 2023

Abstract
TNF receptor-associated factor 6 (TRAF6) is an essential ubiquitin E3 ligase in immune responses, but its function in adaptive immunity is not well understood. Here we show that TRAF6 is recruited to the peripheral ring of the T cell immunological synapse in Jurkat T cells or human primary CD4+ T cells conjugated with SEE-pulsed B cells. This recruitment depends on TRAF6 interacting with linker for activation of T cells (LAT) via its TRAF domain. Although LAT was indispensable for TCR/CD28-induced TRAF6 ubiquitination and its ligase activity, RNA interference-induced TRAF6 knockdown in T cells decreased TCR/CD28-induced LAT ubiquitination, tyrosine-phosphorylation. Overexpression of TRAF6 or its catalytically inactive form C70A promoted and decreased, respectively, LAT tyrosine-191 phosphorylation upon stimulation. Moreover, LAT was ubiquitinated at Lysine-88 by TRAF6 via K63-linked chain. In addition, TRAF6 was required for and synergized with LAT to promote the TCR/CD28-induced activation of NFAT. These results reveal a novel function and mechanism of TRAF6 action in the TCR-LAT signaling pathway distinct from its role in TCR-induced NF-κB activation, and indicate LAT also play an adapter role in TCR/CD28-induced activation of TRAF6.
Effects of sitagliptin on immune function of CD26/DPP-IV in T helper cells (P1190)
Authors: Yoon Hyunyee , Yang Man Gil , Seo Yu Rim , Sung Ji Hyun , Cho Su Hee , Lee Jae Eun , Song Moon Jung 
Published: January 2023

Abstract
CD26 is a multifunctional protein containing an ecto-domain with dipeptidyl peptidase (DPP-IV) activity and known to involve in T cell immune response. A membrane form of DPP-IV (mDPP-IV) is mainly expressed on the surface of T cells, while soluble DPP-IV (sDPP-IV) exists at a high level in blood. Sitagliptin, an sDPP-IV inhibitor is currently available for the treatment of the Type 2 Diabetes. However, whether or how the mDPP-IV activity of T helper cells contribute to their immune functions is not known. In this study, we investigated whether sitagliptin inhibits the mDPP-IV of T helper cells and consequently affects their immune functions when stimulated by pokeweed mitogen (PWM). Our results showed that PWM treatment alone strongly stimulated the expression of mDPP-IV and cytokines such as IL-2, IL-10, TNF-α, IFN-γ, IL-13, GM-CSF and MMP9 in CD4+CD26+ H9 cells. Although apoptosis was induced, the overall cell viability was marginally affected in PWM-treated H9 cells. On the other hand, sitagliptin co-treatment potently inhibited the mDPP-IV activity in a dose-dependent manner, but did not affect either the cell viability or cytokine profiling in PWM-treated H9 cells. The results suggest that T cell signaling pathways for cytokine expression function normally upon immune stimulation although sitagliptin effectively inhibits the mDPP-IV of T helper cell.
Modification of glycan chains as a mechanism to regulate SOCE (P1156)
Authors: Kilch Tatiana , Alansary Dalia , Dörr Kathrin , Peinelt Christine , Niemeyer Barbara 
Published: January 2023

Abstract
T cell receptor mediated activation requires the influx of calcium from the extracellular medium. Here, the endoplasmic reticulum (ER) Ca2+ sensor STIM1 reports the decrease in luminal [Ca2+] caused by IP3 receptor activity, clusters and directly binds and activates Orai1 channels, providing Ca2+ influx (store-operated, SOCE) essential for immune cell activation. We are interested in the regulation and modulation of this pathway. Because alterations of glycosylation of several T cell epitopes are implicated in immunsenescence and cancer, we investigated N-glycosylation site mutants of STIM1 and Orai1 and describe a STIM1 mutant (NN/DQ) which results in a strong gain-of-function. The mutant leads to a significant increase in the number of active channels despite a concomitant decrease in Orai1 protein, uncovering a novel CRAC channel regulatory mechanism. While we have already shown that the STIM1 mutant phenotype is mainly due to an increased rate of STIM1 oligomerization, we now investigate the specificity of this effect by analysing Ca2+ influx mediated by Orai2 and Orai3 and also started delineating the mechanism of Orai1 protein reduction. In addition, we are investigating the complex glycosylation pattern of primary human T cells and its effect on SOCE. Our findings indicate that the cell type specific repertoire of N-glycoslytransferases modifies the influence of the Orai1 glycan chains on Ca2+ influx.
Genome-wide analysis highlights the involvement of calcineurin/NFAT pathway signaling in transcriptional elongation in restimulated memory CD8 T cells (P1166)
Authors: Martinez Gustavo , Aijo Tarmo , Cruz-Guilloty Fernando , Ohora Masatsugu , Greenbaum Jason , Peteres Bjoern , Lähdesmäki Harri , Pipkin Matthew , Rao Anjana 
Published: January 2023

Abstract
Memory T cells respond to antigen encounter more robustly than naïve T cells, but the cell-intrinsic molecular mechanisms underlying this accentuated responsiveness are unknown. Here we show that in in vitro-generated memory-like CD8+ T cells, perforin (Prf1) gene transcription in response to secondary stimulation requires all NFAT transcription factors present in immune cells (NFAT1, NFAT2 and NFAT4), activation of the phosphatase calcineurin, and store-operated Ca2+ entry mediated by the ER Ca2+ sensors Stim1 and Stim2. The calcineurin inhibitor cyclosporine A (CsA) blocked transcriptional elongation through the Prf1 gene, with much less effect on recruitment of RNA polymerase II (Pol II) and transcriptional initiation. The generality of this effect was confirmed by genome-wide RNA-sequencing and chromatin immunoprecipitation for Pol II, NFAT1, H3K4me3 and H3K36me3 histone modifications, and cyclinT1, a component of positive transcription elongation factor b (P-TEFb). Of 1340 genes upregulated upon restimulation, 199 genes were CsA-sensitive, with the majority of these genes affected by transcription elongation rather than Pol II recruitment. CyclinT1 binding to the transcription start sites of restimulation-induced genes overlapped with NFAT1 occupancy and was CsA-sensitive. Thus, a substantial fraction of the rapid TCR-induced memory CD8 T cell responses is regulated by transcription elongation via calcineurin/NFAT signaling and recruitment of P-TEFb.
Autoregulatory CD8 T cells modulate CNS autoimmunity by targeting encephalitogenic CD4 T cells (P4130)
Authors: Ortega Sterling , Kashi Venkatesh , Tyler Andrew , Mendoza Jason , Cunnusamy Khrishen , Karandikar Nitin 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is an inflammatory demyelinating disease with a preponderance of CD8 T-cells in central nervous system (CNS) lesions. We have previously shown that myelin-specific autoreactive CD8 T-cells are capable of suppressing MOG35-55 peptide-induced experimental autoimmune encephalomyelitis (EAE), a murine model of MS. We now report that myelin-specific CD8 T-cells can suppress CD4 T-cell-mediated EAE disease. Whereas in CD8-/- mice, increased EAE severity was associated with increased autoreactivity and inflammatory cytokine production by myelin-specific CD4 T-cells, the adoptive transfer of MOG35-55-specific CD8 T-cells was found to be sufficient for disease reversal. These autoregulatory CD8 T-cells required MHC Class Ia (KbDb) presentation both for in vitro activation and in vivo disease suppression. Interestingly, MOG35-55-specific CD8 T-cells could suppress adoptive EAE using wild type MOG35-55-specific CD4 T-cells induced in KbDb-/- recipients mice, suggesting direct targeting of encephalitogenic CD4 T-cells. Mechanistically, autoregulatory CD8 T-cell-mediated suppression is IFN-γ and perforin dependent and can be clinically augmented with in vitro IL-12 modulation. These studies define a novel and unexpected immune regulatory function for tissue-reactive “autoregulatory” CD8+ T-cells that can be harnessed for the development of immunotherapeutic intervention in MS and other autoimmune diseases.
Higher susceptibility to experimental autoimmune encephalomyelitis in Muc1-deficient mice is associated with increased Th1/Th17 responses (P4134)
Authors: YEN Jui-Hung , Xu Shuyun , Ganea Doina , Kim Kwang Chul 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an autoimmune disease of the central nervous system in which dendritic cells (DC) play an important role in the development of inflammatory responses. Recently it has been shown that Muc1, a membrane tethered glycoprotein, has an ability to suppress inflammatory responses in cultured DC. The objective of this study was to investigate the possible involvement of Muc1 in the development of MS using experimental autoimmune encephalomyelitis (EAE) in mice, a widely used animal model of MS. Our results showed that: (1) Muc1-/- mice developed greater EAE severity compared with wild type (wt) mice, which correlated with increased numbers of Th1 and Th17 cells infiltrating into the CNS; (2) Upon stimulation, splenic DC from Muc1-/- mice produced greater amounts of IL-1β, IL-6, and IL-12 but less amounts of IL-10 compared with those from wt mice; and (3) The ability of splenic DC to antigen-specific CD4+ T cells into Th1 and Th17 cells was greater in Muc1-/- mice compared with wt mice. We conclude that Muc1 plays an anti-inflammatory role in EAE. This is the first report demonstrating the possible involvement of Muc1 in the development of MS and might provide a potential target for immunotherapy.
Alternative splicing of FOXP3 controls regulatory T cell function and Th17 differentiation (P4118)
Authors: Mailer Reiner , Joly Ann-Laure , Liu Sang , Andersson John 
Published: January 2023

Abstract
Eukaryotic cells diversify gene function through alternative splicing, i.e. differential assembly of exons to create distinct protein isoforms. Alternative splicing is common for human FOXP3, the key transcription factor of regulatory T (Treg) cells that is indispensable for maintaining immunological tolerance. Previous studies have demonstrated that Treg cells can exclude exon 7 of FOXP3, which encodes for a leucine zipper region that sustains the Treg cell phenotype. Here we demonstrate that patients suffering from Crohn’s disease (CD) have elevated levels of FOXP3 lacking exon 7, which correlates with disease severity. Moreover, by modulating the pattern of alternative splicing towards exclusion of FOXP3's seventh exon we were able to generate interleukin-2 (IL-2) producing Treg cells. FOXP3 is transiently expressed during Th17 differentiation and we found that exclusion of FOXP3's seventh exon promoted Th17 differentiation. Finally, we demonstrate that expression of FOXP3 lacking exon 7 correlates with increased expression of IL-17 in CD patients. In summary, alternative splicing of FOXP3 regulates both the regulatory T cell phenotype and Th17 differentiation.
iVAX: a sophisticated suite of online vaccine design tools (P3269)
Authors: De Groot Anne , Terry Frances , Spero Denise , Moise Leonard , Martin William 
Published: January 2023

Abstract
Emerging and re-emerging infectious diseases represent a significant challenge for next-generation vaccine design and bioterror preparedness. We have composed a suite of online immunoinformatics tools for accelerated design of genome-derived, epitope-driven vaccines generated from protein sequences. Using the Conservatrix algorithm, even the most mutable pathogenic genomes may be probed for highly conserved segments, which are then mapped for T cell epitopes and regions of high epitope density using EpiMatrix and ClustiMer. Pathogen sequences which could potentially elicit an undesired autoimmune or T-regulatory response due to homology with sequences encoded by the human genome are screened out using BlastiMer and JanusMatrix, an improved homology analysis tool examining pathogen/host sequence similarity with respect to the HLA and TCR faces of an epitope. Immunogenic Consensus Sequences are created by EpiAssembler, a tool which optimizes the balance between pathogen and population coverage. VaccineCAD links potential vaccine candidate epitopes into a string-of-beads design while minimizing non-specific junctional epitopes that may be created in the linking process. With proof of principle established in animal models for vaccines against Tularemia, Vaccinia and H. pylori, the iVAX toolkit exemplifies a rapid, efficient, easily accessible and broadly applicable solution to accelerate the development of critically important vaccines for human health and biodefense.
Production of fully human monoclonal antibodies to therapeutic self-antigenic targets (P3285)
Authors: Sutkowski Natalie , Sun Wei , Fernandes Daniel , Rubinchik Semyon 
Published: January 2023

Abstract
Not all autoreactive antibodies are harmful, despite their role in autoimmune disease. Autoantibodies targeting tumor antigens, growth factors and their receptors are currently used therapeutically for treatment of cancer and autoimmunity. Ordinarily, healthy subjects do not exhibit strong secondary antibody responses to self-proteins, because most autoreactive B cells do not receive sufficient T cell help required for the germinal center reaction and for differentiation into IgG secreting cells. Our data demonstrate that rare, potentially therapeutic autoreactive B cells can be efficiently rescued from tonsil with EBV immortalization, and manipulated in vitro to secrete IgG, by supplying factors that mimic antigen stimulation and T cell help. Because this process is performed wholly in vitro, it constitutes a rapid and cost effective technology, resulting in the isolation of fully human antibodies. In addition, antibodies can be generated against highly toxic antigens, since immunization is unnecessary. As proof of concept, we have developed a panel of human monoclonal antibodies targeting the self-tumor antigen nucleolin for use in cancer immunotherapy. Nucleolin is normally expressed in the nucleus of all cells, but it is over-expressed in both the cytoplasm and on the cell surface of tumor cells in the most common forms of breast cancer, making nucleolin a new type of tumor antigen. The antibodies are currently undergoing pre-clinical testing against breast cancer.
Deep-sequencing reveals WHO class-specific urinary microRNAs in human lupus nephritis (P3245)
Authors: Goilav Beatrice , Ben-Dov Iddo , Loudig Olivier , Blanco Irene , Wahezi Dawn , Putterman Chaim 
Published: January 2023

Abstract
Lupus nephritis (LN) is associated with significant morbidity and mortality. microRNAs (miRs) are small, non-coding RNAs that regulate translation. Previous, studies report changes in miR expression in kidney tissue, urine and PBMC that correlate with LN disease activity. However, LN WHO class-specific miRs have not been described. Using deep-sequencing, we aimed to identify WHO class-specific miRs in urine from adult and pediatric patients with biopsy-proven LN. Cell-free urine from adult (n=25) and pediatric (n=8) female patients with WHO class IV and V LN were obtained at time of active disease and during remission. Total RNA was used to prepare small RNA cDNA libraries for sequencing. Multiplexing through sample-specific 3' adapters was applied. Sequence reads were mapped to the human genome and small RNA databases. miRs were quantified by relative read abundance. We obtained reproducible profiles of miRs. In a paired-sample analysis comparing miR abundance in urine of adult and pediatric patients with active class IV versus class V LN, we found significant changes in 6 miRs, including up-regulation of miR-193a-5p, -423, 501-3p, and -874 by 400-1,000x in WHO class IV LN. In conclusion, we detected higher counts of specific miRs in urine of adult and pediatric patients with LN class IV, versus class V. Given the worse prognosis of class IV LN identifying miRs associated with this LN class is an important step in biomarker discovery of this particulary aggressive disease.