bluuebunny/crossref_metadata_2025_split huggingface dataset:
Investigation of humoral immune response towards persisting Epstein-Barr virus infections in multiple sclerosis and chronic fatigue syndrome using peptide microarrays (TECH1P.868)
Authors: Castro Aaron , Pawlowski Nikolaus , Reimer Ulf , Jansong Janina , Löbel Madlen , Ruprecht Klemens , Scheibenbogen Carmen 
Published: January 2023

Abstract
Most humans carry a considerable number of persisting viral infections, frequently in a latent state. Chronic infections may intermittently reactivate especially under immune suppression and play an important role as trigger or cofactor for autoimmune diseases and cancer. Correlation of such infections with diseases is difficult. An analysis of antibody repertoire in chronic infection may provide information about patterns of virus reactivation. Following primary infection the herpes virus 4 (EBV) results in persistent mostly asymptomatic latent infection. About 95% of the adult population are EBV seropositive. EBV infections are known as cofactors for various diseases such as lymphomas, multiple sclerosis and chronic fatigue syndrome. The detailed mapping of humoral immune response in human serum samples allows a high resolution analysis of the antibody repertoire against EBV antigens. We developed a peptide microarray platform with peptide libraries of up to 6900 members. Here we present data using a library of peptide scans through major EBV antigens from incubations with serum samples of healthy human donors, patients diagnosed with chronic fatigue syndrome or multiple sclerosis. Even though there are shared epitopes between the specimens and cohorts, different epitope patterns are observed for a number of antigens and antigen areas. The patterns of antibody responses could be of diagnostic value for multiple sclerosis and chronic fatigue syndrome.
Multiparametric phenotyping and cell health analysis of immune cell sub-populations on a novel microcapillary cytometer (TECH1P.870)
Authors: Gillis Katherine , Khan Asima , Clor Julie , Tyagarajan Kamala 
Published: January 2023

Abstract
Cellular health enumeration and assessment of heterogeneous cellular sample such as whole blood and PBMCs often require many colors to accurately characterize subpopulations and are critical in defining immune cell handling as well as in functional studies ranging from cytokine secretion to proliferation. Traditional multi-color flow cytometers have posed challenges in multi-parametric experiments, managing compensation, and analyzing results. Existing solutions are expensive, cumbersome to run and require extensive training or dedicated personnel. In this study, we will present data from simplified methods allowing identification of multiple subsets including T,B, NK, naïve and memory, monocytes, and granulocytes while simultaneously assessing cell health for studies with activation agents, cytotoxic treatments or culture conditions on a novel instrument platform, the guava easyCyte™ 12. The guava easyCyte™ 12 is a novel small footprint cytometer, with 3 lasers including blue, red and violet. This allows for data on up to 12 parameters in a microplate or tube-based format. The system is based on principles of microcapillary cytometery, provides absolute count and percentage data without external beads, uses low sample volume and uses flexible and intuitive software. The combination of multicolor antibody panel and cell health markers with the powerful instrument platform enables obtaining critical sub-population level information for PBMC and whole blood samples.
A novel method of exploring the interaction mechanism between small molecul Sudan I and antibody binding (TECH1P.871)
Authors: Xuan Jingxiu , Zhang Chongjie , Huang Jian , Wei Dapeng 
Published: January 2023

Abstract
Sudan I is artificial synthesized compound(MW is 248.28 Da). As an illegally food additive, it is one of the potential carcinogens to human health and safety. The molecular modeling techniques was applied to explore the interaction mechanism of the small molecule and relevant McAb and to analyze the McAb’s specificity. The hybridoma of anti-Sudan I was successfully produced. The 3D-structures of anti-Sudan I and Sudan I, Sudan II, Sudan III and Sudan IV were modeling by Gaussian. The binding free energies between anti-Sudan1 and antigen including Sudan I, Sudan II, Sudan III and Sudan IV were separately -7.04 kcal/mol, -7.48 kcal/mol, -5.02 kcal/mol and -4.25 kcal/mol by Autodock 4.1 . The hydrogen bonds of antigen-antibody binding were generated apart from Sudan IV. The pocket size of anti-Sudan I Ab was 5.23 Å×6.64 Å×10.36 Å, and its volume was 359.56 Å3, and Sudan I, Sudan II, Sudan III and Sudan IV volumes were 228.28 Å3, 261.12 Å3, 323.76 Å3 and 356.56 Å3, respectively. Comparing Sudan I, II, III and IV‘s paths, Sudan I, II were similar but Sudan III, IV couldn’t enter unless they induce the change of the antibody’s conformation. From all above results, the anti-Sudan 1 McAb is specific to Sudan I. It is a novel attempt to explore the interaction mechanism between the small molecule and its relevant monoclonal antibody.
Haploid genetic dissection of immune molecule trafficking (TECH1P.836)
Authors: Shen Jingshi , Davis Eric , Dinarello Charles 
Published: January 2023

Abstract
To perform their expected functions, immune molecules must be targeted to specific cellular compartments such as the plasma membrane. Improper protein targeting can lead to major forms of immune disease such as immunodeficiency and persistent inflammation. Our group developed a novel haploid genetics approach in order to unravel protein targeting on a genome-wide scale. We take advantage of human myeloid cell lines (e.g. KBM7 and HAP1) that exhibit a haploid karyotype. Retroviral insertional mutagenesis of these cells offers an efficient method to generate a library of mutant cells with complete disruption of specific genes. After selections of desired phenotypes (e.g., mislocalization of a protein), the retroviral insertion sites of the mutant clones can be mapped by deep sequencing. We have used this haploid genetics approach to characterize the plasma membrane protein CD59, a key player in the complement system. We identified ~20 genes involved in the biosynthesis, maturation, and trafficking of CD59. These genes are involved in GPI lipid synthesis, oligosaccharide modification, as well as protein trafficking at both the early and late stages of the secretory pathway. This haploid genetic screen provides the first genome-wide map of a complement protein. Many factors identified in the screen represent novel components of the complement system, and may serve as therapeutic targets for treating immune disorders.
ImmuSort, a database on dynamic gene expression and electronic sorting of immune cells (TECH1P.840)
Authors: Wang Pingzhang , Yang Yehong , Han Wenling , Ma Dalong 
Published: January 2023

Abstract
With abundance of accumulated public biological data, it is worthy of consideration about how to fully use these data for wet-experiments. In the current study, we built an immunological database, ImmuSort. ImmuSort provides a convenient way to view global differential gene expression across various experimental conditions or disease states in immune cells, and enables an electronic sorting of these cells in association with a certain expressional intensity. Meanwhile, gene plasticity, comparison of expression level, biomarker discovery and valuation, identification of cell specific transcripts, etc., can also be accessible in the database. Importantly, ImmuSort database can contribute to the discovery of potential novel functional populations of immune cells, e.g., the association of CCL20 bright myeloid populations with inflammation, and provide clues for the function of human and mouse genes in the immune system. ImmuSort is the first product of our immunological database development project. It can be freely accessed on the web at http://202.85.212.211/Account/ImmuSort.html.
High-throughput high-accuracy immune repertoire sequencing (TECH1P.841)
Authors: Jiang Ning , Wu Di , Qu Mingjuan 
Published: January 2023

Abstract
Using VDJ recombination, the human immune system contains hundreds of billions of B and T cells that each expresses a unique antigen receptor through a gene recombination mechanism. This “immune repertoire” forms the base of a safety net that protects each individuals against infections and cancers, with particular lymphocytes proliferating in response to antigenic stimulation. Recently developed immune repertoire sequencing technology has enabled us to quantitatively study the immune repertoire in response to vaccination. However, high rate of errors generated from sequencing the PCR amplification process imposed a challenge on accurate determining repertoire diversity and somatic hypermutations. Recently, we developed a RNA barcoding technology that significantly reduced the error rate from 10-3 to 10-5 nt. This new method greatly improved the over-all diversity and mutation estimates and enabled us to answer many questions in human natural infections.
Impact of IL-2 and IL-10 on human CD4/CD8 T cell composition and Vβ repertoire in a mouse xenogenic GVHD model (TRAN1P.873)
Authors: Abraham Sojan , Choi Jang Gi , Manjunath N. , Shankar Premlata 
Published: January 2023

Abstract
High level of IL-10 is present in sera of patients with Graft-versus-host disease (GVHD) following allogenic bone marrow transplantation. Our studies with NSG mouse model of GVHD induced by transfer of human PBMCs also revealed high levels of IL-10 in the sera, particularly in presence of IL-2, which accelerates the disease. Blocking antibody to IL-10 was unable to protect the animals from GVHD progression. To directly test the effect of IL-10, GVHD was induced after hydrodynamic injection of plasmids encoding human IL-10 singly or in combination with IL-2. When administered alone, IL-10 treatment resulted in more severe GVHD manifestation. Further, in IL-2 accelerated GVHD, IL-10 was unable to ameliorate the course of the disease. There was a marked difference in the composition of the repopulating human T cells in blood and spleen of mice treated with IL-2 or IL-10 alone compared to a combination of the two cytokines. Singly, both cytokines induced expansion of CD4+ as well as CD8+ T cells with a relatively diverse repertoire but in combination, they induced large oligoclonal expansions of CD4+ T cells with very limited Vβ repertoire and near complete absence of CD8+ T cells. Taken together, our results demonstrate for the first time that IL-10 directly contributes to GVHD pathogenesis. Further, when used in combination with IL-2, IL-10 has a major impact on the repertoire of the responding T cells.
Small peptides elicit anti-tumor effects in CT26 model through blocking PD-L1/PD-1 (TUM2P.900)
Authors: Gao Yanfeng , Liu Beiyuan , Chang Haonan , Li Wenwen , Zhou Yang , Li Guodong , Qi Yuanming , Liu Lei 
Published: January 2023

Abstract
It is reported that PD-L1 plays an important role in the immune tolerance of tumor, which through its interaction with PD-1 to induce the T cell anergy and/or exhaustion. Blockade of PD-1/PD-L1 interaction by monoclonal antibody has achieved great success during clinical trial against cancer. Thus, development of novel compounds targeting PD-1/PD-L1 is considered to be very useful. In the present study, a series of small peptides which could bind to PD-L1 were firstly screened out by using phage display peptide library. Several peptides showed potent binding affinity towards PD-L1 and could compete with PD-1 protein to bind PD-L1 expressed on 293T cell surface. In the mouse CT26 colon carcinoma model, these peptides elicited potent anti-tumor activity when injected intratumorly. They could also prolong the survival rate with less effect on bodyweight loss compared to chemotheraputic agent 5-FU. Whether these peptides elicit anti-tumor effects through restoring the T cell function are now being investigated. These peptides could be used as novel anti-tumor agents combined with chemotherapy and radiotherapy.
Modulation of macrophage responses by aberrant lipoproteins in chronic inflammatory diseases. (HUM3P.241)
Authors: Smith Carolyne , Vivekanandan-Giri Anuradha , Playford Martin , Berthier Celine , Kretzler Matthias , Mehta Nehal , Pennathur Subramaniam , Kaplan Mariana 
Published: January 2023

Abstract
Lupus (SLE) patients are fifty times more likely to experience a cardiovascular event compared to age- and sex-matched controls. High levels of oxidized HDL (oxHDL) in SLE patients may contribute to this association. Under normal conditions, unoxidized HDL induces an anti-inflammatory response within macrophages (MØ). We addressed whether lupus oxHDL modifies this response and putative mechanisms associated with enhanced inflammation. Methods: Affymetrix gene expression analysis was performed on control MØs exposed to control or lupus HDL before an LPS challenge. Confirmation at the mRNA and protein level was performed. Signaling pathways involved in gene induction abnormalities were explored using molecular approaches. Results: MØs exposed to SLE HDL up-regulate IL-6, IL-12B and TNF-α at the mRNA and protein levels, compared to MØs exposed to healthy HDL. SLE HDL showed impaired induction of ATF3 mRNA and nuclear translocation. ATF3 activity was restored when the oxHDL binding protein LOX-1 receptor was inhibited with blocking antibodies. SLE HDL also showed a decrease activation of kinases upstream of ATF3 activation. Conclusion: Lupus HDL induces inflammatory responses in macrophages which may promote phenotypic proatherogenic changes and increase cardiovascular risk in this disease. Modifying the HDL proteome in SLE may decrease this risk.
Dysregulation of micro-RNAs involved in TLR pathway in peripheral blood mononuclear cells and synovial fluid mononuclear cells of patients with enthesitis related arthritis form of juvenile idiopathic arthritis (HUM3P.254)
Authors: SINGH SUSHMA , Aggarwal Amita 
Published: January 2023

Abstract
Toll like receptor (TLR) pathway plays a major role in inflammatory arthritis. MicroRNA (miR)-146a, miR-155, miR-21 regulate downstream molecules of TLR signaling like IRAK-1 and TRAF-6. TLR pathway genes are generally upregulated in ERA-JIA. We studied expression of miR-146a, miR-155, miR-21, miR-210 and miR-26a in PBMC and SFMC of patients with ERA. RNA was isolated from PBMC (n=21) & SFMC (n=11) of ERA patients and PBMC of controls (n=11). The miRNA and target gene expressions were analysed by TaqMan qRT-PCR assays. Among 21 patients, 20 were boys. Mean disease onset age was 10.3Y & disease duration was 6Y. All had arthritis, 14 had enthesitis, 4 had uveitis, 7 had IBP and 2 had family history of HLAB27 related disease. No expression difference was seen in miR-146a, miR-155, miR-210 and miR-26a in PBMC of patients and controls but miR-21 was increased in patients. To check inflammatory site specific expression, comparison of SFMC and PBMC was done showing downregulation of miR146a and upregulation of miR-155, miR-21 and miR-210 in SFMC. Expression of miR-26a did not show any change. MiR-146a target genes i.e. IRAK-1 and TRAF-6 were upregulated in SFMC compared to PBMC. There is dysregulation of miRNAs regulating TLR pathway in mononuclear cells of patients with ERA-JIA especially at the chronic inflammation site. Down regulation of miR-146a, a key regulator may lead to over-expression of TLR downstream molecules like IRAK-1 and TRAF-6 thus perpetuating inflammation in ERA.
Expansions of salivary gland CD4+ T cells from Sjögren’s syndrome patients: single-cell repertoire analysis and correlation with clinical measures of disease (HUM3P.255)
Authors: Joachims Michelle , Leehan Kerry , Lawrence Christina , Pan Zijian , Radfar Lida , Rasmussen Astrid , Scofield R. , Sivils Kathy , Thompson Linda , Farris A. 
Published: January 2023

Abstract
Sjögren’s syndrome is a systemic rheumatic disorder characterized by dry eyes, dry mouth and T cell infiltration of exocrine gland tissue. To determine the extent of salivary gland (SG) CD4+ memory T cell clonal expansion and whether these expansions are as frequent in peripheral blood (PB), a multiplex PCR method was used to amplify both the α and β T cell receptor (TCR) sequences from memory CD4+ T cells sorted from SG lip biopsy tissue and PB of 10 primary Sjögren’s syndrome patients. Over 3,000 TCR sequences were obtained from 50-115 (median 91) individual SG and 75-121 (median 104) individual PB T cells per patient. The percentage of cells that were part of clonal expansions was significantly higher in SG (median 11%, range 0-28%) compared to PB (median 0.9%, range 0-7%, p=0.003). Sequence analysis revealed: 1) highly homologous CDR3s among different expanded SG T cell clones within single patients, suggesting antigen-driven expansion and 2) unique cases of convergent recombination among unrelated patients, where different V segments/additions/deletions were utilized to make identical CDR3 amino acid sequences. The percentages of clonally expanded SG T cells correlated significantly with degree of SG fibrosis and reduced saliva production but not with systemic features of disease. SG clonal expansions detected in this study likely identify T cells involved in recognition of common antigen(s) and glandular dysfunction.
A combined genome-wide DNA methylation and transcription analysis identified genes related with pathogenesis of systemic sclerosis (HUM3P.257)
Authors: Li Quanzhen , Zhu Honglin , Luo Hui , Zuo Xiaoxia , Chen Tao , Raman Indu 
Published: January 2023

Abstract
Systemic sclerosis (SSc) is an autoimmune disease with unknown etiology. Studies have revealed epigenetic modification can contribute to susceptibility and pathogenic process in systemic autoimmune diseases by regulating gene activation or silencing. In this study, a genome-wide DNA methylation and transcription analysis was performed on PBMC from 20 SSc patients and 23 matched healthy controls (HC) using the Illumina Human Methy450 and HumanHT-12 Beadchips. Out of 485,000 CpG sites tested across the genome, 4848 hypomethylated and 216 hypermethylated CpG sites were identified from SSc as compared with HC, representing 2488 hypomethylated and 134 hypermethylated genes. Transcriptional microarray analysis identified 1023 significantly upregulated genes in SSc, of which 405 genes were hypomethylated, indicating a negative regulation of DNA methylation on gene expression. The hypomethylated genes in SSC include established autoimmune-related and novel genes, such as AIF1, ARID3A, IFI44L, PARP9, RSAD2, EPSTI1, EIF2AK2 and CYFIP2. Pathway analysis revealed enrichment of genes involved in wnt signaling pathway, leukocyte transendothelial migration, cell adhesion molecules and chemokine signaling pathway. The Wnt signaling pathway was represented by hypomethylation of Wnt5b, Wnt7a, Smad2, and Mapk9. The hypomethylation of genes involved in chemokine signaling pathway were correlated with the increased level of chemokines such as eotaxin, MCP-1 and RANTES in sera of SSc patients.
Obesity-induced inflammatory profile changes in mice and humans (HUM3P.263)
Authors: Mirsoian Annie , Erdembileg Annurad , Sckisel Gail , Berglund Lars , Murphy William 
Published: January 2023

Abstract
Obesity is increasingly becoming a pandemic affecting populations of all ages. Hallmark to obesity is a self-sustaining chronic proinflammatory response characterized by increased immune infiltrates into adipose tissue. However, most studies focus on the impact of metabolic factors upon disease risk. There is currently a paucity of data characterizing the activation and exhaustive phenotypes of immune infiltrates as impacted by obesity. Additionally, there is currently little to no data examining the immunological phenotype of young obese populations, which may provide therapeutic targeting for early disease prevention. In this study, we examined the inflammatory phenotype of lean versus obese mice as well as humans across increasing ages. We observed that increased acute-phase protein levels in young obese cohorts are similar in level to aged cohorts. Our results demonstrate for the first time that immunological changes induced by aging are similarly induced in young obese cohorts. Cytokine analyses demonstrate changes in magnitude of systemic IL-6 levels and PD-1 and Lag-3 expression on CD4 and CD8 T cell populations among young lean, obese, and aged. Importantly, we demonstrate that characterizing immunological systemic changes may provide a better scoring system over metabolic or vascular markers to account for early disease development, and may also provide therapeutic targets for early prevention of age-associated diseases.
CD40 expression on CD4+ T cells (Th40 cells) as a biomarker for multiple sclerosis (HUM3P.265)
Authors: Wagner David , Vaitaitis Gisela , Waid Dan 
Published: January 2023

Abstract
Multiple Sclerosis is a neurodegenerative disease with autoimmune components. Diagnosis is invasive and time consuming. The etiology is unclear as are clues for disease progression. The autoimmune components have largely been defined in the EAE mouse model. The problem has been defining a reliable biomarker for pathogenic cells. We described a unique T cell subset, Th40 cells that is highly auto-aggressive. We compared Th40 levels in clinically isolated syndrome (CIS), relapsing/remitting (RRMS), secondary progressive (SPMS) and primary progressive (PPMS) patients to non-autoimmune controls. MS subjects have significantly increased percentages of Th40 cells compared to controls. In CIS and RRMS Th40 cell levels cover a broad range while in SPMS and PPMS the range is centralized. Longitudinal studies show that Th40 cell levels remain elevated. Th40 cells produce IFNg, TNFa and IL-17. HLA haplotypes were diverse with only 50% of patients expressing the DR2 (DR15) haplotype or DR3 and DR4. Th40 cells are significantly elevated in MS regardless of HLA haplotype and responded to CNS antigens creating antigen signatures. The ratio of Th40 to or to Tr1 cells is 7 to 1. Th40 cells have a memory phenotype. The chemokine receptors CCR5 and CXCR3 were detected. Samples from patients on disease-modifying-therapy (DMT) demonstrated lymphopenia, but the Th40 percentage was not altered. Th40 cells provide a unique diagnostic tool for MS with significant improvement over current options.
Nedd4 family interacting protein 1 limits effector CD4 T cell abundance, cytokine production and inflammation. (HUM3P.248)
Authors: Layman Awo Akosua , Oliver Paula 
Published: January 2023

Abstract
When effector T cells fail to appropriately terminate signaling responses, autoimmune diseases such as multiple sclerosis and inflammatory bowel disease may occur. Signaling in a cell may be terminated by the ubiquitylation and degradation of key signaling transducers. The ubiquitylation cascade is a well studied sequence of orchestrated reactions by which ubiquitin is added on to a substrate protein to, among other things, target that protein for degradation. Ndfip1, or Nedd4 family interacting protein 1, activates the function of E3 ligases, which play important roles in the ubiquitylation cascade. Ndfip1 works with Itch and Nedd4-2 E3 ligases to degrade the IL-4 transcription factor, JunB. Thus in mice that lack Ndfip1, there is unrestricted IL-4 secretion, which leads to an inflammatory phenotype. However, this fails to explain why human SNPs in Ndfip1 have been linked to two autoimmune conditions where TH1 and/or TH17 effector function is unrestrained, namely, multiple sclerosis and inflammatory bowel disease. Mice lacking Ndfip1 show increased overall frequencies of activated T cells and of IFNγ+ and IL-17A+ CD4 T cells, which suggests a general defect in limiting effector T cell abundance. We present data indicating that Ndfip1 limits inflammation in two models of inflammatory bowel disease. Understanding how Ndfip1 limits effector T cell function could clarify how human diseases such as inflammatory bowel disease and multiple sclerosis develop.