bluuebunny/crossref_metadata_2025_split huggingface dataset:
Multiple paths to immunodominance: dominant epitopes are captured by MHC II prior, or post antigen proteolysis, and are enriched by HLA-DM (APP2P.103)
Authors: Sadegh-Nasseri Scheherazade , Kim AeRyon , Hartman Isamu , Cuidad M. Teresa , Song Nianbin , Boronina Tatiana , Caspi Rachel , Jaraquemada Dolores 
Published: January 2023

Abstract
Immunodominance is defined as the responsiveness of T cells to few selected peptide epitopes from complex antigens. To understand the mechanism of epitope selection from antigenic proteins for helper T cells, we used a cell-free antigen processing system composed of five purified proteins: HLA-DR, cathepsins B, H, S, and HLA-DM. We find that immunodominant epitopes are selected differently if derived from self proteins or from pathogens. Sensitivity to cathepsins and resistance to DM-mediated dissociation are equally contributing factors. Self-derived epitopes appear to be less susceptible to cathepsin digestion, whereas pathogen-derived epitopes are sensitive. Cathepsin sensitivity necessitates capture of epitopes by MHC II prior to processing by cathepsins. Resistance to DM-mediated dissociation increases longer epitope survival. We propose that most peptides generated from proteins do not gain immunodominance because they are sensitive to both DM and enzymatic digestions. Thus, immunodominance is established differently for different antigens incorporating functions of DM, cathepsins and MHC II, altogether help to ascertain higher abundance of the immunodominant epitope. Supported by grants from NIAID, R01AI063764 and R21 AI101987 to SS-N.
Evaluation of the role of type I interferon to the development of type 1 diabetes. (BA3P.131)
Authors: Carrero Javier , Calderon Boris , Ferris Stephen , Unanue Emil 
Published: January 2023

Abstract
Our goal is to identify the earliest immunologically relevant events that trigger autoimmune diabetes. The first detectable inflammatory signature in the NOD mouse is the upregulation of type I interferons and an interferon-inducible gene signature. We reason that understanding the early interferon signature may provide targets for the prevention, treatment, or diagnosis of autoimmune diabetes. The two ubiquitous interferon receptors (IFNAR and IFNGR) upregulate hundreds genes, some shared and some unique to each receptor. We backcrossed the type I interferon receptor onto NOD background to determine the relative contribution of IFNAR-inducible transcripts in type I diabetes induction. In our specific pathogen-free colony, NOD.IFNAR-/- mice developed diabetes at a decreased rate and penetrance from littermate controls. This reduction in incidence was gender independent and NOD.IFNAR heterozygous mice developed diabetes normally. There was a reduction in the number of the immunological infiltrates in the islets of Langerhans of NOD.IFNAR-/- mice from 4-8 wks of age as detected by flow cytometry and immunofluorescence. Concomitantly, there was a reduction in inflammatory gene expression in NOD.IFNAR-/- when compared to controls. Additionally, analysis of the NOD.IFNGR-/- mice showed a decrease in penetrance and kinetics of type I diabetes. Based on this data, we hypothesize that IFNAR and IFNGR induce a common genetic signature that is required for type I diabetes to proceed.
Apics deficiency reveals a role for a long noncoding RNA in dendritic cell function and autoimmunity (BA3P.203)
Authors: Brodnicki Thomas , Ashton Michelle , Elso Colleen , Mackin Leanne , Chu Edward , Payne Natalie , Ford Sharon , Tan Iris , Papenfuss Anthony , Kitching A Richard , Bernard Claude , Shortman Ken , Morahan Grant , O'Keefe Meredith 
Published: January 2023

Abstract
Although in vitro observations indicate that long noncoding RNAs (lncRNAs) regulate innate immune responses, their effect upon immune tolerance in vivo has not been defined. We have identified a novel gene of unknown function for which sequence variation is associated with autoimmune diabetes in the nonobese diabetic (NOD) mouse strain. Bioinformatics and expression analyses indicate this gene encodes a lncRNA that is induced by Toll-like receptor (TLR) activation, localizes to the nucleus and cytoplasm of dendritic cells, and binds to proteins within these cellular compartments. Moreover, sequence variation for this gene is associated with altered TLR-mediated cytokine production. Hence this gene was named Apics for Attenuator of Pattern recognition receptor-Induced Cytokine Secretion. To further investigate the function of this lncRNA, we established a C57BL/6 (B6) knockout mouse strain for Apics and discovered that Apics-deficient dendritic cells exhibit enhanced TLR-mediated cytokine production. Apics-deficient B6 mice also exhibit increased susceptibility to autoimmunity in two disease models: experimental autoimmune encephalomyelitis and experimental anti-neutrophil cytoplasmic antibody-associated vasculitis. Our study suggests that lncRNAs, such as Apics, can serve as TLR-inducible repressors that regulate the magnitude of innate immune responses to reduce the risk for developing autoimmune disease.
Disease activity in lupus correlates with expression of the transcription factor ARID3a (BA2P.119)
Authors: Ward Julie , Rose Kira , Montgomery Courtney , Adrianto Indra , Smith Kenneth , James Judith , Merrill Joan , Webb Carol 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a complex autoimmune disease with striking clinical, immunologic and genetic heterogeneity making development of targeted treatment strategies challenging. Previously, we showed transgenic mice with constitutive expression of the DNA-binding protein ARID3a/Bright in all B lymphocytes spontaneously produced autoantibodies. Therefore, we asked if ARID3a expression differed in SLE versus healthy control B lymphocytes. Of 114 SLE patients, 43% had expanded numbers of ARID3a+ peripheral blood B cells. A longitudinal study revealed that numbers of ARID3a+ B cells fluctuated with time, but were highly associated with increased disease activity. ARID3a expression was not limited to B cell subsets that normally express it, but was also apparent in large numbers of naïve B cells in many SLE patients. Therefore, we hypothesize ARID3a+ naïve B cells are precursors of autoantibody producing B cells. Antibody repertoire analyses of ARID3a+ and ARID3a- naïve B cells are in progress. Together, these findings implicate ARID3a as a potential biomarker for disease activity in SLE.
A balance between BCR and inhibitory receptor signaling controls plasma cell differentiation by maintaining optimal Ets1 levels (BA2P.120)
Authors: Luo Wei , Mayeux Jessica , Gutierrez Toni , Getahun Andy , Müller Jennifer , Tedder Thomas , Chen Shu-Hsia , Rickert Robert , Nitschke Lars , Cambier John , Satterthwaite Anne , Garrett-Sinha Lee Ann 
Published: January 2023

Abstract
Signaling through the B cell receptor (BCR) can drive B cell activation and contribute to B cell differentiation into antibody-secreting plasma cells. The positive BCR activation signal is counterbalanced by a number of membrane-localized inhibitory receptors that limit B cell activation and plasma cell differentiation. Deficiencies in these negative signaling pathways may cause autoantibody generation and autoimmune disease in both animal models and human patients. We have previously shown that the transcription factor Ets1 can restrain B cell differentiation into plasma cells. Here, we tested the roles of the BCR and inhibitory receptors in controlling the expression of Ets1 in B cells. We found that Ets1 is down regulated in B cells by BCR or TLR signaling through a pathway dependent on PI3 kinase, Btk, IKK2 and JNK. Deficiencies in inhibitory pathways, such as a loss of the tyrosine kinase Lyn, the phosphatase SHP1 or membrane receptors CD22 or Siglec-G, result in enhanced BCR signaling and decreased Ets1 expression. Restoring Ets1 expression in Lyn- or SHP1-deficient B cells inhibits their enhanced plasma cell differentiation. Our findings suggest that the maintenance of Ets1 expression is an important function of the inhibitory Lyn -- CD22/SiglecG -- SHP1 pathway in B cells.
Dosage of X-linked Toll-like receptor 8 determines gender differences in the development of systemic lupus erythematosus (BA4P.216)
Authors: Umiker Benjamin , Andersson Shauna , Fernandez Luis , Korgaokar Parimal , Larbi Amma , Pilichowska Monika , Weinkauf Craig , Wortis Henry , Kearney John , Imainshi-Kari Thereza 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease with a high incidence in females and a complex phenotype. Using 564Igi mice, a model of SLE with knock-in genes encoding an auto-reactive anti-RNA antibody, we investigated how expression of Toll-like receptors (TLRs) in B cells and neutrophils affects pathogenesis. We established that TLR signaling through MyD88 is necessary for disease. Autoantibody was produced in mice with single deletions of Tlr7, Tlr8 or Tlr9. Autoantibody was produced in female mice with the combined deletions of Tlr7 and 9, but not in male mice. Autoantibody was not produced in the combined absence of Tlr7 and 8 in either sex, indicating that TLR8 contributes to the break in tolerance in females. Furthermore, TLR8 was sufficient for the loss of B cell tolerance, the production of class-switched autoantibody, heightened granulopoiesis, and increased production of type I interferon (IFN-I) by neutrophils as well as glomerulonephritis and death. We show that dosage of X-linked Tlr8 plays a major role in the high incidence of disease in female mice. In addition, we show that the negative regulation of disease by TLR9 is exerted primarily on granulopoiesis and IFN-I production by neutrophils. Collectively, we suggest that individual TLRs play unique roles in the pathogenesis of SLE, suggesting new targets for treatment.
Rapid migration of lymphocytes via murine spleen (CAM1P.226)
Authors: Ganusov Vitaly 
Published: January 2023

Abstract
Spleen is the major secondary lymphoid organ in mammals playing an important role in initiation of adaptive immune response to blood-borne pathogens. Yet, the kinetics of lymphocyte recirculation via spleen in resting and inflammatory conditions remain poorly defined. By perfusing blood and lymphocytes via murine spleen in vitro Ford (Cell Tissue Kinetics 1969) measured accumulation of lymphocytes in circulation over the time of perfusion. I developed a novel mathematical model of lymphocytes recirculation via spleen and fitted that model to Ford's experimental data. The model explains well the majority of experimental data and predicts the average residence time of thoracic duct lymphocytes in the murine spleen of 3 hours. The model predicted that antigenic stimulation of the spleen induces increased migration into the spleen (or increased retention in the spleen) of blood lymphocytes, while cells that recently exited the spleen were nearly not affected. Interestingly, the model was not able to accurately explain the kinetics of lymphocyte migration via the irradiated spleen suggesting that irradiation impacts lymphocyte migration kinetics in a not easily predictable way. Taken together, our analysis provides important insights into kinetics of migration of lymphocytes via spleen.
IL-12p35 single chain subunit antagonizes IL-6 mediated STAT1 and STAT3 activation and prevents expansion of Th17 cells in experimental autoimmune uveitis (CCR1P.246)
Authors: Dambuza Ivy , Yu Cheng-Rong , Wang Ren-Xi , Kim Sung-Ye , Mahdi Rashid , Dolinska Monika , Sergeev Yuri , Wingfield Paul , Egwuagu Charles 
Published: January 2023

Abstract
The Interleukin 12 (IL-12) family of cytokines is comprised of heterodimeric proteins including IL-12 (p35/p40), IL-23 (p19/p40), IL-27 (IL27p28/Ebi3) and IL-35 (p35/Ebi3). IL-27p28 and IL-12p40 have recently been shown to antagonize IL-27 and IL-12 functions, respectively, suggesting that single chain IL-12 family members may act as dominant-negative inhibitors of IL-12 family heterodimeric cytokines. Here, we have genetically engineered IL-12p35 (rIL-12p35) and examined whether IL-12p35 has biological activities independent of its partner, IL-12p40 or EBI3. We show that rIL-12p35 blocked IL-6 mediated activation of STAT1 and STAT3 pathways of TCR-activated CD4+ T cells and inhibited the expansion of Th17 but not Th1 cells. Of particular significance, rIL-12p35 protected mice from autoimmune uveitis by inhibiting pathogenic Th17 cell expansion. Our study demonstrates that IL-12p35 can be used to modulate IL-6-mediated signaling pathways and treat a CNS autoimmune disease.
How can high school students combine computational biology tools with a laboratory model to analyze changes in the human gut microbiome? (EDU1P.247)
Authors: Cobb Andrea , Gillevet Patrick 
Published: January 2023

Abstract
The human gut microbiome is an organ of the immune system. Early in life, an infant and its microbiome interact in poorly understood ways to develop gut-associated lymphoid tissues and to distinguish self from non-self. The gut microbiome produces metabolites used by the host and protects the host from colonization by pathogens. Gut mucosal microbiome composition is fairly constant, but lumen microbiome varies with environmental input. As an AAI Teacher Fellow in Dr. Gillevet’s lab, I used computational tools to determine correlations in alcohol intake, liver health and microbiome composition with changes in inflammatory cytokines and chemokines. How might my students engage in microbiome research? They will construct a gut microbiome model to generate data for computational analysis using Winogradski columns. The columns are simple containers containing wet soil, sulfur and carbon sources. With incubation, a bacterial gradient is established modeling a gut cross section -from lumen to mucosa-column bottom to top -low [O2] and high [S-2]. Students will add an environmental change into columns (for example, alcohol, fiber, fats or antibiotics). After column equilibration, students will collect visual data, then remove column samples to isolate sample DNA and perform 16SrRNA LHPCR and sequencing. Students will use computational analysis of the LHPCR data and (if available) sequence data to generate hypotheses relating environmental input and observed microbiome changes.
STAT5 antagonizes the pre-BCR-dependent NFκB and FOXO tumor suppressor pathways in acute lymphoblastic leukemia (HEM2P.261)
Authors: Katerndahl Casey , Willette Mark , Heltemes Lynn , Ramsey Laura , Koeuth Thearith , Gillespie Emily , Farrar Michael 
Published: January 2023

Abstract
Mice expressing a constitutively active form of STAT5b (Stat5b-CA mice) develop pre-B leukemia with low frequency (~1-2%). Microarray analysis of these leukemias shows decreased expression of genes involved in pre-BCR signaling. Stat5b-CA mice crossed to mice with defects in a coherent signaling pathway downstream of the pre-BCR involving BLNK, BTK, or PKCβ developed pre-B leukemia in &gt; 75% of progeny. These leukemias exhibited reduced expression of a subset of NFκB target genes, which was important for transformation as Stat5b-CA x Nfkb1-/- mice also developed pre-B leukemia. Our results point to an unexpected role for NFκB1 as a tumor suppressor in pre-B cells. Microarrary analysis revealed that STAT5 activation resulted in reduced expression of several NFκB and FOXO target genes, thereby antagonizing these tumor suppressor pathways. To examine this regulation in further detail, we performed ChIP-seq analysis, which showed that STAT5 binds 38% of NFκB target genes that are expressed in pre-B cells. We also observed binding of STAT5 to several FOXO targets including: Foxo1, Cdkn1a, Cdkn2d, Blnk, Vegfa, Cxcr4, Bim, and Dgka. We have validated STAT5 binding to the FOXO targets Foxo1 and Cdkn2d and the NFκB targets Nfkb1, Rel, Irf4, Irf8, Ikzf1, and Ikzf3 by ChIP-qPCR. Our findings demonstrate that STAT5 activation antagonizes the pre-BCR-dependent NFκB and FOXO tumor suppressor pathways that limit pre-B cell proliferation and promote pre-B cell differentiation.
The transcriptional repressor NKAP is required for iNKT cell development and maturation (HEM3P.274)
Authors: Thapa Puspa , Seaburg Lauren , Chen Meibo , McWilliams Douglas , Tangen Sarah , Heibert Scott , Sant' Angelo Derek , Shapiro Virginia 
Published: January 2023

Abstract
Invariant Natural Killer T (iNKT) cells are an innate lineage of lymphocytes that can produce copious amount of cytokines within hours of stimulation. In the thymus, positive selection into the iNKT cell lineage occurs at the double positive (DP) stage, before maturation through various stages (stage 0-3) to complete development. NKAP, a transcriptional repressor, is required for conventional T lymphocyte development and maturation. NKAP interacts with Hdac3 for its repressor function. Previously, using CD4-cre NKAP cKO mice, we have shown that NKAP is required for positive selection into the iNKT cell lineage. To study the role of NKAP in later developmental stages, we generated PLZF-cre NKAP cKO mice with NKAP deletion occurring after entry into the iNKT lineage at stage 0. In the PLZF-cre NKAP cKO mice we observed a significant decrease in the absolute number of iNKTs with a 20-fold reduction at stage 2 and 200-fold reduction at stage 3. This defect is cell intrinsic and cannot be rescued by over-expression of Bcl-XL. Our preliminary results indicate that there may be a block in iNKT cell maturation, as expression of transcription factors Tbet, Gata3, and ROR-γt that define NKT1, NKT2 and NKT17 lineages respectively, are decreased in PLZF-cre NKAP cKO. PLZF-cre Hdac3 cKO mice have a similar defect in iNKT development. Thus, NKAP together with Hdac3 may regulate iNKT cell development and maturation.
CCR2+ monocytes contribute to hematoma clearance and long-term functional recovery after experimental intracerebral hemorrhage (HUM1P.324)
Authors: Hammond Matthew , Taylor Roslyn , Ai Youxi , Sansing Lauren 
Published: January 2023

Abstract
CCR2+ inflammatory monocytes worsen early neurological injury in mouse models of intracerebral hemorrhage (ICH), but their potential roles during recovery are unclear. The purpose of this work was to determine how CCR2+ monocytes influence repair and recovery after ICH. All mice in this study were subjected to the blood injection ICH model, whereby 25 μl of blood were injected into the striatum, a brain region involved in motor coordination. To first determine if monocytes have the capacity to phagocytose debris in the brain, we added 1-μm carboxylate beads to the injected blood. Using an ImageStreamX MarkII imaging flow cytometer, we found that CCR2+ monocytes engulf more beads than resident microglia at day 1 (4.35 ± 2.23 vs. 0.23 ± 0.45 beads, p&lt;0.0001), suggesting they are phagocytically active in the brain after ICH. Next, Ccr2-/- bone marrow was transplanted into CD45.1 hosts to create chimeras with normal microglia, but few circulating monocytes. Controls received C57BL6 marrow. Ccr2-/- chimeric brains contained significantly more residual hemoglobin 7 days after ICH than controls (8.92 ± 1.03 vs. 3.96 ± 2.80 μl, p&lt;0.05), suggesting a role for CCR2+ monocytes in ICH clearance. Importantly, Ccr2-/- bone marrow chimeras exhibit severe motor deficits following an LPS challenge 14 days after ICH, whereas controls exhibit minimal deficits. Together, these data suggest CCR2+ monocytes are important regulators of hematoma clearance and are necessary for functional recovery.
Thy1 (CD90) regulates adipogenesis and is decreased by the environmental obesogen tetrabromobisphenol-A (HUM2P.339)
Authors: Flores ELissa , Woeller Collynn , Phipps Richard 
Published: January 2023

Abstract
Obesity has risen drastically over the last 30 years and occurs through excessive adipogenesis (adipocyte formation) or increases in adipocyte size. Adipocytes produce cytokines such as IL-6 that promotes inflammatory disease. Environmental toxicants termed obesogens such as Tetrabromobisphenol-A (TBBPA) disrupt the endocrine system and increase adipogenesis; however the mechanisms remain unknown. Thy1 (CD90), a cell surface protein, is a member of the immunoglobulin family. We have previously shown that fibroblasts are heterogeneous for Thy1 and only Thy1-/low fibroblasts form adipocytes. We hypothesized that TBBPA promotes adipogenesis by decreasing Thy1 expression. We used mouse 3T3-L1 pre-adipocytes as a model to determine if TBBPA decreases the expression of Thy1 to promote fat cell formation. We measured Thy1 mRNA levels by qPCR and Thy1 protein levels by western blot. Adipogenesis was measured by inspection of lipid droplets and expression of the key adipocyte marker, FABP4. Our data show that TBBPA decreased the expression of Thy1 in 3T3-L1 cells. Furthermore, TBBPA treated cells also expressed higher levels of FABP4, supporting the concept that loss of Thy1 alters cell fate. TBBPA exposure for one week before adipogenic induction resulted in a further reduction of Thy1 and additional increases in FABP4. Thus, prolonged or developmental exposure to TBBPA may affect Thy1 levels and alter physiology long after exposure, which may contribute to the obesity epidemic.
Transgenic mice whose γ 1 and κ genes have been replaced by human ε and κ genes (HYP3P.402)
Authors: Lu Chien-Sheng , Hung Fu-Hsin , Lin Chien-Jen , Chen Jiun-Bo , Chen Christina , Shiung Yu-Yu , Tsai Ching-Yen , Chang Tse-Wen 
Published: January 2023

Abstract
IgE is a primary mediator in most allergic diseases. Human IgE specific for various allergens are very important reagents in diagnostic assays for the management of allergic diseases. However, allergen-specific human IgEs are present in patients’ blood at very low levels and hence extremely difficult to prepare. We have now constructed a transgenic homozygous mouse strain, referred to as HεκKI strain, whose γ1 constant region gene segment has been replaced by human ε gene constant region segment and whose κ constant gene replaced by human κ constant gene. In those HεκKI mice, the human IgE levels are about 10 times of mouse IgE. Upon immunization with an allergen papain, both human IgE and mouse IgE were increased, with human IgE increased to about 30 folds of mouse IgE. We have prepared a number of hybridomas secreting monoclonal human IgE (the variable regions being mouse) specific for papain. Cells of a basophilic leukemic line pulsed with such human IgE could be activated by papain to undergo degranulation process. These results indicate that the transgenic mouse strain HεκKI could be employed to prepare polyclonal and monoclonal human IgE specific for various allergens.
Role of mast cells in a chronic murine asthma model (HYP3P.348)
Authors: Song Chang Ho , Zhang Xin , Chai Ok Hee , Shon Dong-Hwa , Lim Jeong Min , Kim Ik Seong , Han Eui-Hyeog , Kim Hyoung Tae 
Published: January 2023

Abstract
Mast cells are thought to be important in the cause of allergic diseases, but the role of mast cells in the airway remodeling of chronic long-term asthma remains controversial. In this study, using the mast cell “nock in” strategy to get the mast cell-deficient WBB6F1-W/Wv mice (W/Wv) selectively reconstituted bone marrow-cultured mast cells (BMCMCs) from normal congenic wild-type mice (+/+) (+/+ BMCMCs→W/Wv mice), mice were employed to study the roles of mast cell in a murine asthma model. The mice were sensitized with ovalbumin (OVA) and challenged with OVA for 14 weeks. The mice were assessed airway reactivity by PenH. The lungs and trachea were stained with H-E for pathologic alteration, toluidine blue for mast cell, congo red for eosinophil, PAS for goblet cell and Masson’s-trichrome for fibrosis. Mast cells were not observed in the lungs of control and OVA-sensitized and challenged W/WV mice, but OVA-sensitization and challenge increased mast cell number in +/+ mice and +/+BMCMCs→W/Wv mice. OVA-sensitization and challenge could enhance airway reactivity, pathologic alteration, eosinophil infiltration, goblet cell hyperplasia and fibrosis in +/+ mice. The pathologic changes of OVA treated W/Wv mice were less than that of +/+ mice, but +/+BMCMCs→W/Wv mice could restore the reaction to OVA-sensitization and challenge of +/+ mice. These results indicate that the mast cell play an important role in the OVA-induced chronic murine asthma model.