bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow cytometry based analysis for subsets of Plasmacytoid Dendritic Cells (pDC) from human PBMC (100.31)
Authors: Chitta Sriram , Quintel Payton , Lee Hyun-Ku , Wardle Lauren , Stampfl Jason , Singh Gita , Rosenberg Jonathan , Singh Sujay 
Published: January 2023

Abstract
Dendritic cells are a heterogeneous group of antigen presenting cells that link innate and adaptive immunity. In mice and humans, subsets of DC have been identified, based on origin, location, differentiation, phenotype and functions. Newly identified markers, availability of mAb(s) and advanced flow-cytometry techniques have helped in enumerating existence of such subsets. Plasmacytoid dendritic cells (pDC) are peripheral blood DC subsets, which produce IFN (Type I) and are vital for anti-viral immunity. pDC exist in small numbers (~0.1-0.5%) in peripheral blood and their phenotype analysis by flow, includes CD123 and HLADR expression on lineage negative populations of PBMC. Recently, pDC were shown to play important roles (besides anti-viral) in other human diseases such as lupus and psoriasis but it is not known if distinct subsets are associated. In the present study we used three markers viz. CD123, BDCA-2 and ILT7 (newly identified) to analyze potential subsets of pDC. As Type I IFN producers, pDC express intracellular toll-like receptors TLR7 and TLR9 and our analysis extends to include whether these subsets differ in TLR expression, particularly focusing on TLRs playing an important role in innate-IFN (Type I) production. In order to understand the role of pDC (subsets) in induction of anti-immune responses towards other microbes, expression of non-specific markers such as Dectin-2 and TLR 10 were also tested using newly available mAb(s).
Uptake and disposition of recombinant chimeric immunogens constructed from Streptococcus mutans SBR-CTA2/B at mucosal immune inductive sites (100.35)
Authors: Zhao Weiwei , Minderman Hans , O'Loughlin Kieran , Russell Michael 
Published: January 2023

Abstract
Purpose: To investigate cellular pathways of uptake and processing of the recombinant chimeric immunogen SBR-CTA2/B consisting of the saliva-binding region (SBR) of S. mutans AgI/II coupled to cholera toxin A2/B subunits, in comparison with SBR alone. Methods: Amnis ImageStream technology was used to characterize cells from lamina propria (LP), Peyer’s patch (PP), and mesenteric lymph nodes (MLN) of mice before and 2, 4, 16, and 48h after intragastric immunization with FITC-labeled proteins. Dendritic cells (DC) were stained for phenotypic markers and the migration receptor CCR7 using contrasting fluorochromes. Results: LP CD103+, CD11c+, and especially CD103/CD11c double+ DC took up SBR-CTA2/B through 4h, after which the proportion of antigen-containing DC stayed level until 16h, although the number of DCs continued to increase. DCs in PP captured SBR-CTA2/B through 16h. The proportion of LP DCs containing SBR peaked at 2h and declined by 16h. CCR7 expression by CD11c+ LP DCs taking up SBR-CTA2/B rose at 2h and dropped at 4h, whereas CCR7 expression by DC taking up SBR declined from 0h levels. In MLN the number of CD11c+/CD4+ cell doublets dramatically increased at 48 h after SBR-CTA2/B immunization. Conclusions: Compared to SBR alone, the chimeric protein SBR-CTA2/B enhanced the functional activation of DCs in LP and PP, in which most antigen uptake occurred within the first few hours. DCs loaded with protein may then migrate to MLN and intercommunicate with T cells.
Immunoproteasomes regulate gene expression in dendritic cells (100.36)
Authors: de Verteuil Danielle , Laperrière David , Mader Sylvie , Thibault Pierre , Perreault Claude 
Published: January 2023

Abstract
A constitutive form of the proteasome (CP) is expressed in all eukaryotic cells and is involved in many cellular processes. Immune and cytokine-activated cells also express a second type of proteasome named the immunoproteasome (IP). Since its discovery, it has been thought that the main role of IPs is to produce MHC I-associated peptides. However, an increasing number of studies report peptide-independent effects of IPs. In line with this, we recently showed using microarrays that knocking-out two IP-subunits (LMP7 and MECL1) in bone marrow-derived dendritic cells alters the expression of 171 genes mostly associated with immune functions, with enriched clusters on chromosomes. We now show by qPCR experiments that the disruption of both LMP7 and MECL1 is necessary to alter gene expression. Bioinformatic predictions of transcription factor binding sites suggest that this effect of IPs hinges mainly on transcriptional regulation. Comparing the abundance of unspliced and mature mRNAs in WT and IP knock-out cells shows that IPs also influence the maturation of certain transcripts. However, IPs do not affect mRNA stability overtime when blocking mRNA synthesis. The control of gene expression by IPs might explain aberrant T and B cell phenotypes previously described in IP-subunit knock-out mice. A better comprehension of how IPs regulate gene expression could help us understand the selective pressure that allowed the conservation of more than one type of proteasome in vertebrates.
Targeting Antigens to Human Dendritic Cells via Dectin-1 Elicits Potent Antigen-specific Th17 CD4+ T Cells That Express Mucosal Homing Receptors (100.39)
Authors: Ni Ling , Li Xiaohua , Duluc Dorothee , Joo HyeMee , Kluca Peter , Agouna-Deciat Olivier , Zurawski Sandy , Banchereau Jacques , Zurawski Gerard , Oh SangKon 
Published: January 2023

Abstract
Emerging evidence indicates important roles of IL-17-producing (Th17) CD4+ T cells in protective immunity against microbial pathogens, including influenza viruses. Thus we tested the magnitudes and spectrum of influenza viral hemagglutinin subunit 1 (HA1)-specific Th17 CD4+ T cells in healthy donors by targeting HA1 to dendritic cells (DCs) via Dectin-1 using recombinant fusion proteins of agonistic anti-Dectin-1 fused to HA1 (anti-Dectin-1-HA1). Our data show that healthy individuals maintain broad ranges of influenza HA1-specific Th17 CD4+ T cells expressing CCR4 and CCR6. Interestingly, HA1-specific Th17 CD4+ T cells were distinguished from other Th17 CD4+ T cells by expressing increased levels of CCR9 and decreased levels of CD161. We further found that DCs activated through hDectin-1 upregulated other mucosal homing receptors, beta7 integrin, CCR4, and CCR9 on the surface of HA1-specific Th17 CD4+ T cells. Such HA1-specific Th17 CD4+ T cell responses were greatly enhanced by targeting DCs with anti-Dectin-1-HA1 in the presence of TLR2 ligands. This enhancement was mainly due to the expansion of pre-existing HA1-specific memory Th17 CD4+ T cells that express IL-1R1, but not naïve CD4+ T cells which do not. Induction of IL-1R1 on naïve CD4+ T cells required for priming HA1-specific Th17 CD4+ T cells. Taken together, we conclude that antigen targeting to DCs via Dectin-1 could mount potent protective immunity against microbial pathogens, including influenza viruses.
Insulin peptide specific CD4+ T cells escape negative selection and cause diabetes in NOD mice (100.44)
Authors: Mohan James , Calderon Boris , Petzold Shirley , Herzog Jeremy , Unanue Emil 
Published: January 2023

Abstract
The importance of insulin as an autoantigen in type 1 diabetes pathogenesis has been extensively associated with the development of disease. Within the NOD mouse the majority but not all of autoreactive T cells that recognize insulin are deleted during development in the thymus. We found a set of autoreactive CD4+ T cells specific for an insulin peptide, incapable of recognizing the insulin protein when processed by the antigen presenting cells (APC). These unique autoreactive cells escape thymic control, infiltrate the islets of Langerhans in pre-diabetic mice and are pathogenic. The mechanism by which these peptide specific T cells recognize antigen results from the formation of a unique state of the peptide-MHC complex (pMHC) that differs from the pMHC generated from the processed protein. Differences in pMHC states of the same peptide bound to the MHC molecule may arise from the particular vesicle within the APC were exogenous peptide vs. processed protein is loaded. We have also found that APC situated in close contact with the beta cells in the islets of Langerhans bear vesicles with the antigenic insulin peptides and activate the peptide-specific T cells. The importance of these T cells is that they readily escaped negative selection when the insulin protein was expressed by the thymic APC even though they can be activated in the periphery by islet APC in the absence of inflammation.
Analysis of several CD4+ T cell clones reveals a &gt;10,000 fold range in 2D affinity of antigen recognition (100.45)
Authors: Hood Jennifer , Sabatino Joseph , Rosenthal Kristen , Zhu Cheng , Evavold Brian 
Published: January 2023

Abstract
Using a micropipette adhesion frequency assay, we are able to determine the affinity of polyclonal and monoclonal T cells without needing to isolate the TCR. This assay allows for the direct measuring of adhesion between a T cell and a surrogate APC, which is a human red blood cell that has been coated with peptide:MHC monomers. Utilizing this two dimensional measure of affinity, we determined the effective affinities for the following CD4+ TCR transgenic clones: 2D2, BDC2.5, and SMARTA for their cognate antigens. For the 2D2 cells, we assessed both MOG38-49:IAb and NF-M18-30:IAb monomers since it has been shown that 2D2 cells can cross-react with both of these CNS antigens. Our results demonstrate a wide range of affinities for these various transgenic cells. The hierarchy of the affinities from highest to lowest for the clones (clone:peptide) was; BDC2.5:1040-63, SMARTA:GP-61, 2D2:NFM18-30 and 2D2:MOG38-49 and roughly correlated with their responsiveness to antigen. The affinities ranged from 10-4 µm4 to less than 10-8 µm4, which is below the limits of detection for this technique. These data demonstrate that there is a wide range of affinities for CD4+ T cells that are capable of responding to antigen.
Modulation of MHC class I antigen presentation by the adenovirus E3-19K protein (100.8)
Authors: Fu Jie , Li Lenong , Bouvier Marlene 
Published: January 2023

Abstract
The presentation of viral antigens by MHC I to CTLs is vital for elimination of infected cells. In turn, viruses have evolved various strategies to abrogate antiviral cellular immune responses. For example, the adenovirus (Ad) E3-19K protein retains MHC I in the endoplasmic reticulum, dramatically suppressing the cell-surface presentation of viral antigens. Many aspects of how E3-19K exerts its function towards MHC I are poorly understood. This knowledge is important as the E3-19K/MHC I interaction is thought to play a role in enabling Ads to cause persistent infections. We characterized biophysically and cellularly interaction between E3-19K proteins of different Ad serotypes (Ad 7 and 35, subgroup B; Ad 2 and Ad 5, subgroup C; Ad 37, subgroup D; and Ad 4, subgroup E) and HLA-A, -B, and -C molecules. Our results show that E3-19K proteins from Ad serotypes of the same subgroup display more similar binding properties for a given MHC I than those of different subgroups. Furthermore, E3-19K proteins exhibited allele- and locus-specificity; higher avidity for HLA-A relative to -B molecules, and no interaction with HLA-C molecules. FACS analysis showed that binding affinities correlated (in a negative way) with levels of MHC I expression on infected cells. Finally, we propose the first model of interaction between E3-19K and MHC I. Overall, our studies provide novel insights into the structure/function relationship of E3-19K and the molecular cell biology of antigen presentation.
Serologic analysis of MHC class I folding intermediates (100.9)
Authors: Simone Laura , Tuli Amit , Wang Xiaojian , Sharma Mahak , Simone Peter , Naslavsky Naava , Caplan Steve , Solheim Joyce 
Published: January 2023

Abstract
During the process of peptide selection, MHC class I molecules transition from an open, peptide-free conformer to a folded, peptide-bound state. Thus, MHC class I molecules have been proposed to adopt an intermediate conformation during peptide binding; however, the molecular nature of this transition state is unknown. Here, we report evidence that the 34-1-2 antibody, which recognizes several murine MHC class I molecules, is able to distinguish an intermediately folded conformer. The majority of 34-1-2+ Ld molecules were found to represent a distinct pool of MHC class I molecules that were not recognized by the anti-folded Ld antibody 30-5-7. Moreover, 34-1-2+ Ld molecules were found to display characteristics indicative of incomplete folding, including increased tapasin association and endoplasmic reticulum (ER) retention. Additionally, our data suggest that tapasin associates with sub-optimally loaded 34-1-2+ Kd molecules and may facilitate the retrieval of these molecules to the ER. We have designated this MHC class I conformation as 34-1-2-INT (34-1-2+, intermediately folded). Together with prior knowledge regarding the 34-1-2 epitope, our analysis supports a model whereby binding of the peptide N-terminus permits MHC class I molecules to adopt an intermediately folded conformation which subsequently converts to the fully folded form upon optimization of C-terminal interactions. Thus, the 34-1-2 antibody is an important tool in understanding MHC class I peptide loading.
TCR tuning by homeostatic cytokines — implication for autoimmunity (101.16)
Authors: Deshpande Pratima , Singh Karnail , Weyand Cornelia , Goronzy Jorg 
Published: January 2023

Abstract
In animal models, lymphopenia and the associated function of homeostatic cytokines (HC) set the stage for autoimmunity. This model is relevant for rheumatoid arthritis (RA) which has defects in T cell homeostasis due to a failure in DNA repair and which responds to JAK inhibition. We have previously shown that RA T cells have a hyperreactive ERK response which impairs the induction of anergy. Here, we examined whether HC tune TCR activation thresholds by calibrating the ERK rheostat. RA T cells exhibited increased levels of phosphorylated STAT3 and STAT5 indicating in vivo activity of IL-21/IL6 and IL-7/IL-15. Incubation of healthy donor T cells with IL-7, IL15 or IL21 prior to anti-CD3/CD28 stimulation augmented ERK activation and expression of activation markers (CD69, CD40L) and cytokines (TNFα, IFNγ and IL-17). Preincubation with IL-1β or TNFα did not exert an effect. In DR4 healthy donors, HC priming facilitated responses to self-antigens such as citrullinated vimentin peptide. The effect of HC lasted for less than 3 hours and was sensitive to PI3K inhibition suggesting a non-transcriptional PI3K mediated priming of the ERK pathway. HC dose titration showed a bimodal distribution in pERK consistent with an on-off switch characteristic of a positive feedback loop. Allosteric binding of RAS-GTP to SOS is known to induce such an on-off switch. Indeed, HC incubation increased active RAS. We propose that HC lower the threshold for autoimmune responses by priming of SOS.
Expansion of IL-7Ralow memory CD8 T cells in human lupus (101.19)
Authors: Lee Naeun , Shah Kamini , Shin Min Sun , Kim Sang Hyun , Craft Joe , Kang Insoo 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an inflammatory autoimmune disorder that is characterized by production of autoantibodies and immune complex deposition. In the immunopathogenesis of SLE, CD4+ T cells play as critical drivers of B cell dependent autoantibody response, but little is known about the role of CD8+ T cells in the development of SLE. Recent studies have shown that the expansion of memory CD8+ T cells occurs in SLE patients. We have measured the expression of IL-7 receptor alpha (IL-7Ra, CD127) on memory CD8+ T cells in SLE patients, since IL-7 and IL-7R are involved in homeostasis of CD8+ T cells. As previous reported, we found two subsets expressing high and low levels of IL-7Ra on memory CD8+ T cells in SLE patients and healthy controls. In this study, we showed that SLE patients have an increased frequency of IL-7Ralow memory CD8+ T cells compared to healthy controls. This cell subset expresses high levels of cytotoxic molecule perforin with the capacity to damage tissue. Furthermore, the frequency of IL-7Ralow memory CD8+ T cells correlated with disease activity as measured by SLE disease activity index (SLEDAI). Although T cell receptor triggering barely induced IL-7Ralow cell proliferation, the homeostatic cytokine IL-15 substantially induced the proliferation of this cell subset. Our findings suggest a potential role for IL-7Ralow memory CD8+ T cells in the development of SLE.
Activation of human macrophages by the endogenous toll-like receptor 4 ligand high molecular weight hyaluronan (101.20)
Authors: Wallet Mark , Guiulfo Giorgio , Wallet Shannon , Sleasman John , Goodenow Maureen 
Published: January 2023

Abstract
Rheumatoid arthritis [RA] is caused in part by cytokines secreted by classically activated synovial macrophages. We reported that IFNγ, typically produced by CD4 T cells within synovitis, primes macrophages for activation by high molecular weight hyaluronan [HMW-HA], a major component of joint synovial fluid. HMW-HA is a ligand for both toll-like receptor 4 [TLR4] and CD44. The purpose of this study was to compare the molecular mechanism of HMW-HA (endogenous TLR ligand) versus LPS (exogenous TLR ligand) induced macrophage activation. Transcriptome bioprofiles were determined by cDNA arrays, and the roles of CD44 and TLR4 were determined using blocking antibodies and cytokine measurement by Milliplex®. Significant differences in gene expression profiles were revealed when comparing IFNγ priming versus mock priming under both LPS and HMW-HA activating conditions, although nearly identical gene expression profiles were observed when comparing HMW-HA to LPS stimulation under both mock and IFNγ priming conditions. Blockade of CD44 enhanced inflammatory cytokine secretion while blockade of TLR4 inhibited inflammatory cytokine secretion in response to either HMW-HA or LPS. HMW-HA provides the stimulus required for classical macrophage activation when cells are primed with IFNγ in a response that is essentially identical to the LPS response. Type 1 T cells in proximity to endogenous TLR ligands may provide sufficient signals to initiate classical macrophage activation in RA.
Pivotal role of dermal γδ T Cells in the pathogenesis of psoriasis (101.24)
Authors: Cai Yihua , Yan Jun 
Published: January 2023

Abstract
Interleukin (IL)-23 and CD4+ Th17 cells are thought to be critical in the development of psoriasis. Here, we report that IL-23 predominantly stimulates innate dermal γδ T cells to produce IL-17 that leads to disease progression. Dermal γδ T cells constitutively express high levels of IL-23R, transcriptional factor RORγt, and various chemokine receptors. However, dermal γδ T cells are CD27 negative and do not produce IFN-γ. IL-17 production from dermal γδ T cells is independent of αβ T cells. The epidermal hyperplasia and inflammation induced by IL-23 is significantly decreased in TCR δKO and IL-17RA KO mice but occurs normally in TCR αKO mice. The clinical manifestations including epidermal thickening and neutrophil infiltration in an Imiquimod (IMQ)-induced mouse model of psoriasis are also significantly decreased in TCR δKO mice. Perhaps further promoting disease progression, IL-23 preferentially stimulates dermal γδ T cell expansion in vitro. In human psoriasis patients, γδ T cells are also greatly increased in affected skin and similarly produce large amounts of IL-17. Thus, IL-23-responsive dermal γδ T cells are the major IL-17 producers in the skin and may represent a novel target for the treatment of psoriasis.
Eosinophils promote myocarditis and dilated cardiomyopathy by enhancing Th17 responses (101.27)
Authors: Baldeviano Geral , Barin Jobert , Wu Lei , Talor Monica , Ferrer Patricia , Chen Ping , Legault Jillian , Ong Sufey , Zheng Dongfeng , Bedja Djahida , Gabrielson Kathleen , Chatzidimitriou Dimitris , Rose Noel , Cihakova Daniela 
Published: January 2023

Abstract
Eosinophilic myocarditis is a rare but potentially fatal form of myocarditis that is occasionally associated with systemic eosinophilia. It is not known how eosinophils localize to the myocardium or which are the mechanisms underlying their involvement in inflammatory heart diseases. Here, we assessed the role of eosinophils using a murine model of experimental autoimmune myocarditis (EAM). We have previously observed that eosinophils are recruited to the inflamed heart. Eosinophil-deficient mice (ΔdblGATA1) had decreased EAM and did not develop dilated cardiomyopathy (DCM) compared to wild-type BALB/c control mice. In contrast, IL-5 transgenic mice (IL-5Cd3) developed peripheral eosinophilia, severe EAM, increased myocardial fibrosis and accelerated DCM. Transfer of bone marrow-derived eosinophils increased EAM severity in ΔdblGATA1 mice, suggesting a direct pathogenic role for eosinophils in myocarditis. The trafficking of eosinophils to the heart was independent of IL-5 and CCR3 but partially dependent on the cytosolic phospholipase-2. Flow cytometry analysis revealed that the frequency of IL-17A-secreting T cells tracked with the number of eosinophils; it was significantly increased in IL-5Cd3 and decreased in ΔdblGATA1 mice. This study provides the first direct evidence for a pathogenic role in eosinophils in myocarditis and DCM. Additionally, our findings support the hypothesis that eosinophils may promote Th17 responses during myocarditis.
Opposing roles for Fas ligand and IL-10 in regulating insulitis initiation (101.32)
Authors: Hamad Abdel Rahim , Uddin Sophia , Marshall Andrew , Nakata Chiaki 
Published: January 2023

Abstract
Type 1 diabetes (TID), leading autoimmune disease of childhood, is caused by destruction of beta-cells by autoreactive T cells. Despite daily insulin injections, patients often develop cardiovascular complications and there is a need for identifying therapeutic targets to prevent the disease without impinging on host defense. Fas ligand (FasL) can be such a potential target. Fas/FasL interactions primarily regulate T cell homeostasis but not activation. Nevertheless, mutation of FasL (gld) prevents autoimmune diabetes in NOD mice, the widely used model for T1D. Although homozygous gld mutations cause age-dependent lymphoproliferation, limiting the gld mutation to one allele (NOD-gld/+) or treating NOD-wt mice with FasL-neutralizing mAb inhibits insulitis and prevents disease development without causing lymphoproliferation or immunosuppression. Here we show that the gld mutation did not inhibit proliferation and expansion of diabetogenic T cells in pancreatic lymph nodes but inhibited their accumulation in the pancreata. A significant proportion of B cells found in pancreata of NOD-gld/+ mice surface-expressed CD5 and produced IL-10. Neutralization of IL-10 allowed CD4+ T cell accumulation in the pancreas and development of insulitis in NOD-gld/+ mice. The results provide novel mechanistic insights into the disease pathogenesis with potential implications for developing novel T1D therapy.