bluuebunny/crossref_metadata_2025_split huggingface dataset:
DEC-205-mediated antigen targeting to steady-state dendritic cells induces deletion of cognate CD8+ T cells independently of the PD-1 pathway (123.19)
Authors: Mukherjee Gayatri , Geliebter Ari , Sharpe Arlene , Freeman Gordon , DiLorenzo Teresa 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is an autoimmune disease characterized by T cell-mediated destruction of the pancreatic islet beta cells. Cytotoxic CD8+ T cells specific for beta cell antigens are a main contributor to T1D development. CD8+ T cells specific for insulin and islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) epitopes have been implicated in T1D development in both patients and NOD mice. Targeted delivery to steady-state dendritic cells (DCs) of a beta cell peptide mimotope linked to anti-DEC-205 leads to deletional tolerance of transferred cognate CD8+ T cells in NOD mice. Treatment with anti-DEC-205 linked to NRP-V7, a superagonist mimotope of IGRP206-214, selectively deleted endogenous IGRP206-214-specific T cells, a highly prevalent specificity in NOD mice, and showed promise in T1D prevention. Simultaneous treatments with anti-DEC-205/InsI9, a mimotope peptide of the CD8+ T cell insulin epitope InsB15-23, and anti DEC-205/NRP-V7 are being held to study the effect of targeting multiple CD8+ T cell specificities on T1D prevention. Investigation of the operative tolerance mechanisms revealed that antibody blockade of the PD-1 pathway, often implicated in DC- mediated induction of peripheral tolerance, could not inhibit the deletion of transferred T cells by DEC-205-mediated targeting of their antigen to steady- state DC. Involvement of the CTLA-4 or Fas pathways will be probed to further study the molecular mechanism of tolerance induction. .
Pre-B Cell Leukemia Homeobox 1is a novel susceptibility gene that regulate T cell functions. (123.2)
Authors: Sengupta Mayami , Liang Shujuan , Morel Laurence 
Published: January 2023

Abstract
Pre-leukemia homeobox (Pbx) 1 is a novel lupus susceptibility gene that regulates T cell activation and tolerance in both the NZM2410 lupus-prone mouse model and lupus patients. Lupus susceptibility is associated with the expression of a novel splice isoform Pbx1-d, which, when over expressed leads to an increased CD4+ T cell activation and a reduced induction of Tregs by retinoic acid. The exons encoding for the DNA binding domain and the Hox binding domain are deleted in Pbx-1d, predicting that this isoform functions as a dominant negative. To test this hypothesis, we used an osteogenesis model in which Pbx1 expression prevents differentiation. Pbx1-d expression in MC3T3 osteoblast progenitors mimicked the effect of shRNA Pbx1 in accelerating osteoblast differentiation. Furthermore, Pbx-1d binding to the SOX3 promoter was weaker than that of the full-length Pbx1-b isoform, and SOX3 expression was attenuated in Pbx1-d transfected cells. These results demonstrated that Pbx1-d functions as a dominant negative. C57BL/6 transgenic mice expressing Pbx1-d in their CD4+ T cells show an age-dependent increased in CD4+ T cell activation, with an expansion of CD44+ CD4+ memory cells, and the production of anti-nuclear autoantibodies. These results demonstrate that Pbx1 represents a novel regulator of T cell activation, whose impaired function leads to systemic autoimmunity.
T-regulatory cell (Treg)-extrinsic role of IL-2 in protection against Autoimmune Pancreatitis (123.31)
Authors: Sharma Rahul , Ju Shyr-Te 
Published: January 2023

Abstract
Autoimmune pancreatitis (AIP) is a chronic inflammation of pancreas marked by high serum immunoglobulins and leukocytic infiltration. While the Il2-/- mice, which have reduced Treg levels, develop severe pancreatitis, the Foxp3-mutant Scurfy (Sf) mice, with a complete Treg-deficiency, are resistant to pancreatitis though they have a fatal multiorgan inflammation. Rendering Sf mice deficient for IL-2 (Sf.Il2-/-) abolished this resistance to AIP. We have shown that IL-2 is critical for the Th2 and not the Th1 response in the Sf mice. Thus, we hypothesized that a strong Th2 response in the Sf mice protects against AIP while a poor Th2 response due to IL-2 deficiency fails to suppress the disease. Indeed, Sf mice lacking IL-4 (Sf.Il4-/-) or STAT6 (Sf.Stat6-/-) developed severe AIP, while IFN-γ -deficient Sf mice (Sf.Ifng-/-) remained free from pancreatitis. Interestingly, all these mice had hyper-gammaglobulinemia and serum reactivity to carbonic anhydrase II. Our data further shows that lack of IL-2 or inadequate Th2 expression in the Sf mice causes an increase in the IFN-γ levels leading to upregulation of CXCR5, CXCR3 and CCR5 on the CD4+ T-cells and an increase in IP-10 and RANTES levels locally in the pancreas. The CD4+ T-cells then infiltrate the pancreas and form tertiary lymphoid follicles leading to AIP. We propose that although Treg-deficiency contributes to AIP, IL-2 regulates clinical manifestation of the disease in a Th2 dependent manner.
Cell mediated injury in autoimmune anti-glomerular basement membrane disease is defined by a critical T cell epitope in humanized mice (123.33)
Authors: Ooi Joshua , Chang Janet , Hudson Billy , Fugger Lars , Holdsworth Stephen , Kitching A Richard 
Published: January 2023

Abstract
Human anti-glomerular basement membrane (GBM) disease has strong associations with HLA DRB1*1501. The target autoantigen is the non-collagenous domain of the α3 chain of Type IV collagen, α3(IV)NC1. T cell epitopes of α3(IV)NC1 were identified by immunizing mouse MHC-/-, HLA DRB1*1501 transgenic (1501tg) mice with overlapping 20mers. They defined a HLA DRB1*1501 restricted epitope (aa128-147) not reactive in HLA DRB1*0101 transgenic (0101tg) or C57BL/6 mice. 1501tg mice were immunized with α1(IV)NC1, α3(IV)NC1 or α1/α3 chimeric molecules, made by inserting the α3 aa17-31 (C2) or aa127-141 (C6) into the non-immunogenic α1 backbone. 1501tg mice immunized with α3(IV)NC1 or C6 responded to aa128-147 but not to aa8-27, but α1(IV)NC1 or C2-immunized mice did not respond to either peptide. CD4+ T cell clones, made from 1501tg mice and transferred into naive 1501tg mice, induced disease with necrotizing and crescentic glomeruli, albuminuria and renal failure, and glomerular CD4+ T cell and macrophage infiltrates. To determine if immunization with the restricted epitope causes disease, 1501tg or 0101tg mice were crossed with FcγRIIB-/- mice (Fcγ receptors are implicated in disease susceptibility). Immunization with either whole α3(IV)NC1 or the identified restricted epitope induced glomerulonephritis in 1501tg.FcγRIIB-/- mice but not in 0101tg.FcγRIIB-/- mice. These studies demonstrate that T cell responses in anti-GBM disease are defined by a HLA DRB1*1501 restricted T cell epitope.
CD28-ITK signals regulate autoreactive T cell trafficking into tissues (123.41)
Authors: Jain Nitya , Miu Bing , Jiang Jian-kang , Prince Amanda , Greiner Dale , Thomas Craig , Sanderson Michael , Berg Leslie , Kang Joonsoo 
Published: January 2023

Abstract
Autoimmune diseases such as Type 1 diabetes arise from the activation of self- reactive T cells and their trafficking to target tissues, ultimately leading to impaired organ function. Immune-based therapies for autoimmunity including corticosteroid regimens, modulation of inflammatory effectors and tolerance induction by inhibition of T cell activation fail to target self-reactive T cells specifically, resulting in compromised general immunity. A recent focused approach aims at regulating self-reactive T cell migration to organs to limit immune pathology. The signaling pathways regulating migration of activated tissue-antigen specific T cells versus activated foreign-antigen specific T cells are unknown. Here we demonstrate that the B7-CD28 costimulatory pathway of T cell activation is an important regulator of the trafficking of self-reactive T cells. The Tec-kinase ITK is a downstream mediator of the CD28 signal controlling autoreactive T cell migration. Using the Ctla4-/- mouse as a model of multi-organ autoimmunity, we show that genetic and pharmacological inhibition of ITK prevents the migration of activated T cells into target tissues. Furthermore, small molecule ITK inhibitors prevent pancreatic islet infiltration by diabetogenic T cells in mouse models of type I diabetes. Our results suggest that pharmacological disruption of ITK function should be considered as a treatment of human T cell mediated organ-specific autoimmune diseases.
IL27p28 inhibits induction and expression of experimental autoimmune uveitis by concurrently antagonizing autoaggressive Th1 and Th17 responses. (123.43)
Authors: Chong Wai Po , Horai Reiko , Silver Phyllis , Mattapallil Mary , Chen Jun , Caspi Rachel 
Published: January 2023

Abstract
Human autoimmune uveitis is accompanied by elevated Th1 and Th17 responses. In experimental autoimmune uveitis (EAU) induced in mice with the retinal antigen IRBP, both responses are present and each effector lineage can mediate disease independently. A therapeutic approach that targets both Th1 and Th17 would therefore be attractive. IL27p28 is an antagonist of gp130, which is required for IL-6 and for IL-27 signaling. Since IL-6 and IL 27 promote Th17 and Th1 responses respectively, we hypothesized that IL27p28 would regulate EAU by affecting both responses concurrently. Overexpression of IL27p28 reduced IFN-γ and IL-17A production from eye-infiltrating CD4+ T cells and ameliorated EAU induced by active immunization. Mechanistic studies revealed that IL27p28 suppressed IL6-mediated STAT3 and IL27-mediated STAT1 phosphorylation during Th17 and Th1 polarization, respectively. Furthermore, overexpression of IL27p28 ameliorated EAU induced by passive transfer of uveitogenic effector Th1 or Th17 cells obtained from donors expressing an IRBP-specific TCR. Donor TCR-transgenic T cells recovered from these mice expressed reduced levels of Th17 and Th1 signature cytokines, IL-17A and IFN-γ, and of their corresponding transcription factors, RORγt and Tbet. The ability of IL27p28 to inhibit both Th1 and Th17 responses and to ameliorate not only the induction, but also the effector phase of EAU, bodes well for clinical usefulness of IL27p28 augmentation to control autoimmunity.
CDK2 regulates Foxp3 activity through protein destabilization (123.7)
Authors: Morawski Peter , Mehra Parul , Chunder Neelanjana , Wells Andrew 
Published: January 2023

Abstract
Foxp3 is a transcription factor that directs development of the regulatory T cell (Treg) lineage. While the signals that regulate expression of the foxp3 gene are established, it remains unclear how the activity of the Foxp3 protein is controlled. Examination of the primary structure of Foxp3 revealed four putative cyclin-dependent kinase (CDK) substrate motifs: S/T-P. In vitro kinase and mass spectrometry analyses showed that Foxp3 can be phosphorylated on at least three of these motifs by the E-type cyclin-dependent kinase, CDK2. Ser/Ala and Thr/Ala mutations within these motifs (S/T&gt;A, or STAble) resulted in an increase in the stability, measured through a CHX assay, and steady-state level of Foxp3 protein within the cell. Conventional CD4 cells transduced with the STAble mutant of Foxp3 exhibited increased CD25 expression, repression of IL-2, and suppressive function compared to CD4 cells transduced with wild type Foxp3, suggesting that phosphorylation of Foxp3 by CDK2 may negatively regulate its function. Consistent with this, natural Foxp3+ Tregs from mice genetically deficient for CDK2 likewise exhibited a gain of suppressive function compared to wild type Tregs. Based on these data, we hypothesize that mitogenic signaling through CDK2 negatively regulates Foxp3 activity and Treg function through phosphorylation-dependent protein destabilization.
Direct Effects of Increased Sugar Concentration on Proliferation and Activation of Jurkat T-cells. (123.8)
Authors: Snitsarev Vladislav 
Published: January 2023

Abstract
High-fructose corn syrup has been used with increasing commonality in recent decades. Parallel with its use, an increased occurrence of autoimmune disease has been reported suggesting a potential correlation. We hypothesized that increased sugar concentration may activate T-cells. Using Jurkat T-cells as a model, we found that increased glucose and fructose concentration decreased Jurkat T-cell proliferation and death rates. This effect was significant at pH 7.4, while at pH 7.2 the above effects did not reach significance. Cells grown in high glucose and fructose concentration formed clusters that were unbreakable by standard trituration techniques indicating possible expression of high-affinity adhesion molecules characteristic of T-cell activation; this effect was more prominent in fructose than glucose formulations. We conclude that exposure of T-cells to sustained higher sugar concentration can lead to cells aggregation indicating possible activation in an antigen-independent manner, and can potentially contribute to the development of autoimmune disease.
Rapid Methods for Cell Health Assessment of Peripheral Blood Mononuclear Cells (124.15)
Authors: Clor Julie , Gillis Katherine , Khan Asima , Tyagarajan Kamala 
Published: January 2023

Abstract
Immunological studies evaluating phenotypic, mechanistic and activation characteristics of peripheral blood mononuclear cells (PBMCs) involve multiple samples, long treatment times, and laborious methods. The initial screening and assessment of PBMC quality for parameters such as count, viability and apoptosis is critical in determining the success of downstream immunological studies. Traditionally, such screening has been limited since it has required access to complicated and expensive platforms. In this study, we present results from a simplified method for the characterization of fresh or cryopreserved PBMC’s on the novel Muse Cell Analyzer. The methods utilize simple mix and read assays, provide quick assessment of count and viability measurements and the apoptotic status of PBMC’s. Our results indicate that good accuracy and linearity data on count and viability can be obtained for PBMC’s in a wide concentration range can be obtained using sample sizes as low as 20 µL. Precise results were obtained for all assays tested as shown by the low coefficient of variation (CV&lt;10%). The simplified assay format allows for low operator to operator variability with inter-operator CV’s of &lt;10%. Data on impact of PBMC viability and apoptosis and correlation with downstream performance will be presented. The combination of rapid methods with a novel accessible platform will empower researchers to easily select optimal PBMC samples for downstream experiments.
Cytokine responses and ADCC activity in cryopreserved PBMC (124.6)
Authors: Bahunde Faith , Awoyode Rotimi , Przybylek Andrew , Harwood William , Meixell Ron , Anekella Bharathi 
Published: January 2023

Abstract
Cryopreserved peripheral blood mononuclear cells (PBMC) are a good alternative to fresh PBMC in clinical trials when samples are collected at multiple sites and processed later. Optimal retention of cryopreserved PBMC functionality depends on cell isolation, freezing and storage. SeraCare developed validated PBMC cryopreservation procedures to ensure functionality is maintained on cryopreservation. Two assays were performed to demonstrate the equivalence of cryopreserved vs. fresh PBMCs. Intracellular cytokine staining (ICS) quantitatively measures antigen specific T cell cytokine production at the single cell level. Using flow cytometry ICS analyzes multiple parameters on a single sample to obtain the precise cytokine producing T cell phenotype. Our data demonstrates production of IFN-γ, IL-2, TNF-α and IL-4 from cryopreserved PBMC suggesting full retention of PBMC functionality after cryopreservation. The antibody dependent cellular cytotoxicity (ADCC) assay measures natural killer cell mediated destruction of antibody targeted cells. ADCC is a powerful tool to measure immune responses in HIV and cancer immunotherapy. Most studies used fresh PBMC in ADCC assays, noting that cryopreserved PBMC result in lower activity. We show cryopreserved PBMCs have comparable ADCC activity to fresh PBMC. ADCC activity appeared donor specific and was not affected by cryopreservation. Our data suggest that cryopreservation has no adverse affect on PBMC functionality in immunological assays.
Analysis of T and B cell phenotype and effector memory T cell response in a mouse model of tree nut allergy (125.10)
Authors: Gangur Venugopal , Gonipeta Babu , Parvataneni Sitaram , Duriancik David , Kim Eunjung , Gardner Elizabeth 
Published: January 2023

Abstract
Tree nuts cause potentially fatal anaphylactic reactions for reasons that are not well understood. We previously reported an adjuvant free mouse model of hazelnut allergy. Here we analyzed the T & B cell phenotype and effector memory responses. Balb/c mice were transdermally sensitized with hazelnut. Hazelnut allergy was confirmed by measuring hazelnut specific IgE antibodies and systemic anaphylaxis (hypothermia) upon challenge. Using flow cytometry T cell and B cell phenotypes were studied. Hazelnut allergic mice exhibited a significant increase in the number of CD4+ CD62L - T cells in the bone marrow and the spleen. Among B cells, there was increase in CD80+ cells in BM, spleen and MLN and CD62L- cells in BM and spleen of allergic mice. Among CD80+ B cells a significant proportion were CD73- in the MLN of allergic mice. Effector memory Type-2 cytokine responses were analyzed using spleen cell cultures. Allergic mice exhibited a robust IL-4, IL-5 and IL-13 response that was blocked by anti-CD4 mAb. The STAT6 KO mice exhibited IL-5 and IL-13 but not IL-4 memory response. These data demonstrate that hazelnut elicits: (i) significant expansion of CD4+CD62L- T cells in both the BM and the periphery; (ii) a modest increase in CD80+ and CD62L- B cells in BM and the periphery; and a significant down regulation of CD73 expression on a subset of CD80+ B cells in the MLN; and (iii) STAT6 and CD4 dependent IL-4 memory; and STAT6 independent, CD4 dependent IL-5/IL-13 memory.
LTB4 driven neutrophil recruitment to the skin is essential for allergic skin inflammation (125.17)
Authors: Oyoshi Michiko , He Rui , Li Yitang , Yoon Juhan , Afshar Roshi , Chen Mei , Lee David , Luo Hongbo , Luster Andrew , Larson Allison , Murphy George , Geha Raif 
Published: January 2023

Abstract
Scratching triggers skin flares in atopic dermatitis (AD), a disease marked by itching and allergic skin inflammation. Scratching of human skin, and tape stripping of mouse skin, caused a neutrophil influx. This influx in mice was largely dependent on the generation of leukotriene B4 (LTB4) by neutrophils and their expression of its receptor BLT1. Allergic skin inflammation in response to epicutaneous (EC) application of ovalbumin to tape-stripped skin was severely impaired in BLT1-/- mice, and required the expression of BLT1 on both T cells and non-T cells. Co-transfer of WT neutrophils, but not neutrophils deficient in BLT1 or the LTB4 synthesizing enzyme LTA4H, restored the ability of WT CD4+ effector T cells to transfer allergic skin inflammation to BLT1-/- recipients. Pharmacologic blockade of LTB4 synthesis inhibited allergic skin inflammation elicited by cutaneous antigen challenge in previously EC-sensitized mice. Blockade of the LTB4-BLT1 axis might provide a potential therapy for AD.
Role of carbohydrate moieties in recognition and uptake of allergens by innate immune cells (125.2)
Authors: Ghaem Maghami Amir , Al-Ghouleh Abeer , Harrington Helen , Shakib Farouk 
Published: January 2023

Abstract
Allergens are initiators of both innate and adaptive immune responses. They are recognised at the site of entry by epithelial and dendritic cells (DCs), both of which activate innate inflammatory circuits that can collectively induce Th2 immune responses. In an attempt to have a better understanding of the role of carbohydrates in the recognition and uptake of allergens by the innate immune system, we defined common glycosylation patterns in major allergens like Der p 1, Fel d 1, Ara h 1, Ber e 1, Der p 2, Bla g 2 and Can f 1. Our data showed that all these allergens are glycosylated and that the main dominant sugars on allergens are 1-2, 1-3 and 1-6 mannose. We then investigated uptake and recognition of different allergen glycofroms (i.e. natural, hyper- and a-glycosylated) by DCs and lung epithelia. The deglycosylated preparation of Der p 1 exhibited minimal uptake by DCs compared to the natural and hyperglycosylated recombinant counterparts, with the latter being taken up more readily than the other preparations. Interestingly, our data also showed an increase in Thymic stromal lymphopoietin (TSLP) (known to drive DC maturation in support of Th2 responses) secretion by lung epithelia upon stimulation with natural Der p 1 which was carbohydrate dependent. Collectively, our data indicate that carbohydrate moieties on allergens play a vital role in their recognition by innate immune cells, implicating them in downstream deleterious Th2 cell activation and IgE production.
Tolerance induction using novel antigen-coupled carriers in a model of allergic asthma (125.5)
Authors: Smarr Charles , Yap Woon Teck , McCarthy Derrick , Bryce Paul , Shea Lonnie , Miller Stephen 
Published: January 2023

Abstract
Current immunotherapy for Th2-associated allergic disease is inefficient and has a significant risk of inducing anaphylaxis. There is significant need for new treatments to safely, efficiently induce antigen-specific tolerance in allergic disease. Antigen-coupled cells (Ag-SP) have been used by our lab to prevent and treat Th1/Th17-mediated models of autoimmune disease and, more recently, Th2-mediated models of allergic asthma and food allergy. Ag-SP treatment is currently entering a Phase II clinical trial in multiple sclerosis patients in Germany. Due to the technical complications of such treatment, however, we have begun to explore new antigen carriers for the induction of tolerance in autoimmune and allergic diseases. Initial work with antigen coupled to inert polystyrene and biodegradable PLGA microparticles (Ag-MP) identified these carriers as effective at inducing tolerance in EAE. We have extended these findings to a common ovalbumin (OVA)-alum induced mouse model of allergic asthma. Prophylactically, OVA- MP induce protective tolerance - inhibiting production of OVA-specific IgE antibodies and lung inflammation and preventing OVA-specific proliferation and Th2 cytokine production by restimulated lymphocytes. Given post-sensitization, OVA-MP inhibit Th2 responses in restimulation cultures but do not, in the short frame examined, inhibit other parameters of disease. This work identifies Ag-MP as promising inducers of tolerance for allergic disease.
Retinoic acid downregulates IL-4-induced IgE production through retinoic acid receptor (125.8)
Authors: Seo Goo-Young , Lee Jeong-Min , Yoo Hana , Nagler Cathryn , Kim Pyeung-Hyeun 
Published: January 2023

Abstract
Retinoic acid (RA) has been reported to downregulate IgE Ab synthesis. In the present study, we assessed the mechanisms by which RA suppresses IL-4-induced IgE expression by murine splenic B cells. We found that AM80, a RA receptor (RAR) agonist, inhibits the production of IL-4-induced IgE but not IgA or IgM. Addition of the RAR antagonist LE540 to these cultures abrogated the RA mediated inhibition of IgE, as well as the expression of germ-line IgE transcripts (GLTϵ). In addition RA decreased IL-4/αCD40-induced GLϵ promoter reporter activity while increasing GLα promoter activity. Finally, serum IgE was substantially elevated in vitamin A-deficient (VAD) mice. Taken together, these results suggest that RA downregulates IgE isotype switching through a RAR-dependent pathway in vivo.