bluuebunny/crossref_metadata_2025_split huggingface dataset:
FPLC in a pipette tip (P3297)
Authors: Mundodi Suparna , Rishi Porecha , Murray Anderson , Nicole Rapicavoli 
Published: January 2023

Abstract
Molecular characterization of the immune response and how this process is implemented requires the purification of native proteins and antibodies. There are numerous purification methods to isolate proteins of interest, but these methods usually require multiple steps to attain the level of purity required for most studies. The Rainin PureSpeed Protein Purification System utilizes pipette tips containing purification resin at their distal end interfaced with an E4 XLS pipette to simplify enrichment procedures for virtually all proteins. The E4 XLS pipette, when set up within a 96-deepwell plate, is able to carry out semi-automated purification of antibodies and other native or recombinant proteins. The pipette is able to drive up-and-down sample flow over the immobilized resin matrix for as many as 12 samples in parallel. The ability of the pipette to carry out liquid handling steps enhances laboratory productivity by allowing an investigator to step away from the apparatus until the next step in the protocol. This new, innovative technology from Rainin Instrument is available with pipette tips containing protein A, protein G, Ni-IMAC, or one of four different ion exchange resins, which allow for multiple options for protein purification. Finally, the PureSpeed System functions well in advanced applications such as protein and chromatin immunoprecipitation, offering speed and versatility relative to agarose and magnetic bead technologies.
ES-62 prevents development of lupus-like pathology in the MRL/Lpr mouse (P5210)
Authors: Rodgers David , McGrath Mairi , Pineda Miguel , Al-Riyami Lamyaa , Harnett William , Harnett Margaret 
Published: January 2023

Abstract
The prevalence of autoimmunity in developing countries is lower than that in the developed world and this inverse relationship correlates with the level of chronic parasitic infections. Co-evolution of parasites with the human immune system has resulted in evasion strategies that prevent the host clearing the parasite yet limit pathology. One mechanism involves the release of immunomodulators like ES-62, secreted by the filarial nematode Acanthocheilonema viteae: ES-62 exhibits broad anti-inflammatory activity and therapeutic potential in autoimmune diseases such as rheumatoid arthritis. Here we show that ES-62 protects against nephritis, as indicated by proteinuria levels, in the murine MRL/Lpr model of systemic lupus erythematosus (SLE). It has been proposed that the pro-inflammatory cytokine IL-17 is a primary driver of disease, in mouse models as well as in SLE patients, but our data do not support targeting of IL-17 as the major protective mechanism underlying ES-62 efficacy. Indeed, and consistent with this, we have shown that neutralising antibodies specific for IL-17 do not block development of proteinuria. ES-62 is a large immunogenic glycoprotein and therefore not suitable for therapeutic use. Thus, since the active moiety of ES-62 is phosphorylcholine (PC), we have recently developed small molecular analogues based around PC that mimic the immunomodulatory effects of ES-62 in autoimmune disorders, including SLE.
The filarial nematode product, ES-62 targets IL-22 to mediate protection in collagen-induced arthritis (P5212)
Authors: Pineda Miguel , Harnett William , Al-Riyami Lamyaa , Harnett Margarett 
Published: January 2023

Abstract
Parasitic worms skew immune responses to a Th2-like phenotype, inhibiting Th1/Th17 pathways and expanding regulatory cells such as Tregs. Worms can achieve such immunoregulation by secreting specific products like ES-62, a glycoprotein secreted by the filarial nematode A. viteae. We have previously shown ES-62 to be protective in Collagen-Induced Arthritis, a murine model of rheumatoid arthritis, and that the mechanism associated with protection involves a down-regulation of the cytokine IL-17. We have now analyzed the effects of ES-62 on other Th17-related cytokines focusing on IL-22, a cytokine that has been reported to exhibit both pro and anti-inflammatory actions in animal models of inflammatory disease. In CIA, administration of ES-62 appears to target the IL-17/IL-22 balance in lymph nodes and joints of animals resulting in modulation of the number and inflammatory phenotype of infiltrating and resident cells in the joint, and consequent attenuation of inflammation. Interestingly, using a combination of recombinant IL-22 and blocking antibody in in vivo studies, the precise type of effect of IL-22 on CIA progression was shown to depend largely both on the location of IL-22 action and also the stage at which the cytokine is produced during disease progression. These findings have allowed us to identify a novel mechanism used by the helminth product ES-62 to reduce autoimmune arthritis and they also contribute to our understanding of the complex biology of IL-22
Therapeutic effect of mesenchymal stem cells in combination with oncolytic adenoviruses for the treatment of solid tumors in an immunocompetent mouse model (P2020)
Authors: Rincon Esther , Kanojia Deepak , Auffinger Brenda , UIlya Ilya , Han Yu , Alemany Ramon , Ramírez Manuel , García-Castro Javier , Lesniak Maciej 
Published: January 2023

Abstract
Oncolytic adenoviruses (Ad) constitute a promising alternative for patients with solid tumors. Unfortunately, clinical trials have shown limited efficacy due to insufficient viral delivery to tumor and the ability of Ad to trigger antiviral immune response. We have previously documented enhanced Ad delivery in vivo using human mesenchymal stem cells (MSCs). However, a major limitation in the field is the lack of immune- and replication-competent models. In this study, we used syngeneic CMT64 cells as an immunocompetent model to analyze the antitumoral response followed by treatment with MSCs and oncolytic Ad ICOVIR5. First, we demonstrated that ICOVIR5 replicates and has cytopatic effect in CMT64 cells. To evaluate the in vivo efficacy of ICOVIR5 using MSCs as carriers, CMT64 cells were grown in mouse flanks. Once the tumors were established, mice were divided into 4 groups (n=5/group). Groups 1/2 were treated with saline and groups 3/4 with MSCs. Given the fact that mouse MSCs do not support the replication of human Ad, tumors were injected with saline (groups 1/2) or ICOVIR5 (groups 3/4) to mimic the carrier role of human MSCs. This therapy was performed weekly for 3 weeks and the tumor volumes were measured. A significant decrease in the tumor volume (p&lt;0.05) was found in the group treated with MSCs and ICOVIR5. These results establish a potential permissive model to study immune mechanisms involved in the antitumoral effects of MSCs as carriers for oncolytic Ad.
Viral infection alters subnuclear localization of PD-1 and CD62L loci in LCMV-specific CD8 T-cells (P6369)
Authors: Sacristan Catarina , Youngblood Ben , Xu Jean , Lu Peiyuan , McGary Katelyn , Hewitt Susannah , Skok Jane , Boss Jerry , Ahmed Rafi , Dustin Michael 
Published: January 2023

Abstract
During chronic viral infection, CD8 T-cell differentiation yields a unique transcriptional profile: T-cell exhaustion. The suboptimal effector function of exhausted cells is partly due to sustained expression of PD-1. Using the LCMV mouse model of infection, we examined pdcd1 (PD-1) and sell (CD62L) gene regulation at different stages of T-cell differentiation. We hypothesized that epigenetic programming and nuclear localization of these genes were coupled to viral persistence and T-cell differentiation. Changes in gene expression are linked to locus positioning in specific compartments such as nuclear lamina (silencing). Using DNA-FISH, we report a significant decrease in pdcd1 alleles associated to nuclear lamina in exhausted cells relative to other CD8 subsets. Dissociation from lamin was consistent with upregulated expression of PD-1 mRNA and protein, and with loss of PD-1 DNA methylation in exhausted cells. In contrast, we observed a significant increase in sell recruitment to lamin with exhaustion, mirroring changes in sell gene expression. CD8 T-cells from Blimp-1-deficient chronically infected mice exhibited an impaired ability to regulate pdcd1 and sell association to lamin, thus implicating Blimp-1 in modulating expression of both genes during chronic infection. Our findings contribute to the understanding of the molecular pathways mediating T-cell memory and exhaustion, and may provide insight into implementing therapeutic strategies to resolve chronic infections.
Longitudinal analyses of human sera reveal expansions of antibody titers specific to previously-encountered pandemic and seasonal influenza viruses (P6366)
Authors: Miller Matthew , Gardner Thomas , Krammer Florian , Aguado Lauren , Tortorella Domenico , Basler Christopher , Palese Peter 
Published: January 2023

Abstract
It is known that early exposures with influenza viruses impact the outcome of subsequent encounters. However, how sequential exposures to antigenically variant viruses shape the humoral immune response remains very poorly understood. To address this, a longitudinal analysis of antibody titers against pandemic (Japan/57 H2N2, Hong Kong/68 H3N2 and USSR/77 H1N1) and seasonal (Alabama/81 H3N2 and Texas/91 H1N1) strains of influenza virus was performed on matched serological samples collected over a 20 year period (1987-2008). These samples were obtained from the Framingham Heart Study Offspring Cohort. Sustained increases in neutralizing antibody titers were observed against both the pandemic and seasonal isolates. Hemagglutinin stalk-reactive antibody titers rose modestly over the 20 year study period, even in the absence of major antigenic shifts which are known to boost these antibodies most effectively. Neutralizing antibody titers against cytomegalovirus were also investigated. In contrast to influenza virus, no sustained increase in neutralizing antibody titers specific to cytomegalovirus was observed. These findings represent the first detailed longitudinal analysis of the humoral immune response against influenza virus in humans, and suggest a unique role for sequential exposures to antigenic variants in the maintenance of neutralizing antibodies.
The tapasin-related protein TAPBPR is a new component of the MHC class I presentation pathway (P5000)
Authors: Boyle Louise , Hermann Clemens , Boname Jessica , Rhodes David , Lehner Paul , Trowsdale John 
Published: January 2023

Abstract
Tapasin is an integral component of the peptide-loading complex (PLC) important for efficient peptide loading onto MHC class I molecules. We investigated the function of the novel tapasin-related protein, TAPBPR. Like tapasin, TAPBPR is widely expressed, interferon-γ inducible and binds to MHC class I coupled with β2-microglobulin (β2m) in the endoplasmic reticulum. In contrast to tapasin, TAPBPR does not bind ERp57 or calreticulin and is not an integral component of the PLC. β2m is essential for the association between TAPBPR and MHC class I. However, the association between TAPBPR and MHC class I occurs in the absence of a functional PLC, suggesting peptide is not required. Expression of TAPBPR decreases the rate of MHC class I maturation through the secretory pathway and prolongs the association of MHC class I on the PLC. The TAPBPR:MHC class I complex trafficked through the Golgi apparatus, demonstrating a function of TAPBPR beyond the ER/cis-Golgi. The identification of TAPBPR as a novel component of the MHC class I antigen presentation pathway demonstrates that mechanisms controlling MHC class I expression remain incompletely understood.
Dipeptides enhance folding and peptide binding of “empty” MHC class I molecules (P5002)
Authors: Saini Sunil Kumar , Ostermeir Katja , Ramnarayan Venkat Raman , Zacharias Martin , Springer Sebastian 
Published: January 2023

Abstract
MHC (Major Histocompatibility Complex) class I molecules play a central role in the mammalian antiviral immune response by presenting the viral proteome at the cell surface for recognition by cytotoxic T lymphocytes. The folding of recombinant MHC class I molecules from denatured bacterial inclusion bodies and their assembly with the light chain beta-2 microglobulin largely depend on specific high-affinity peptides of usually eight or nine amino acids. To find the minimum peptide requirement for folding and stabilization of class I molecules, we have investigated the influence of dipeptides on the folding and peptide binding of recombinant HLA-A*02:01 (A2) and H-2Kb (Kb). In the presence of the dipeptide Gly-Leu (GL), both A2 and Kb folded well, even without a high-affinity peptide present, as measured by a tryptophan fluorescence assay. When folded in the presence of GL, they also bound much faster to their respective specific high-affinity peptides as when folded without GL. Kinetic analysis by fluorescence anisotropy revealed that the presence of GL enhances the on-rate of peptide binding to empty A2 and Kb by six to eight fold. Molecular dynamics simulation indicates that the dipeptide GL interacts with the C terminus of the peptide binding groove. In conclusion, our data demonstrate that the dipeptide GL improves the folding of A2 and Kb and induces a peptide-receptive conformation for enhanced peptide binding.
The role of B cells in neuropsychiatric disease in murine systemic lupus erythematosis (P4000)
Authors: Wen Jing , Stock Ariel , Gulinello Maria , Putterman Chaim , Wang Haowei , Shlomchik Mark 
Published: January 2023

Abstract
Neuropsychiatric disease in systemic lupus erythematosus (NPSLE) is a common and dangerous disease manifestation. However, the mechanisms underlying NPSLE are not fully understood, and the optimal treatment is not known. One postulated mechanism involves lupus autoantibodies with affinity for brain tissue, which due to a compromised blood brain barrier that allows for the passage from the serum into the brain can deposit in the CNS to induce cell damage and neuropsychiatric abnormalities. Therefore, B-cell depletion would be expected to be an effective intervention for NPSLE. To address this question experimentally, we studied B-cell depletion in a murine lupus model with prominent neuropsychiatric manifestations, the MRL-lpr/lpr strain (JHD-MRL/lpr). Comprehensive neurobehavioral tests including forced swim, anhedonia, open field, object recognition, object placement, and social preference were employed to evaluate the neuropsychiatric manifestations in B cell depleted JHD-MRL/lpr as compared to B cell sufficient MRL/lpr mice. Unexpectedly, we found that complete B cell depletion did not ameliorate major NPSLE features, including depression-like behavior and cognitive dysfunction. However, improved motor activity was observed in JHD-MRL/lpr mice. Thus, the absence of B cells and autoantibodies is not sufficient to ameliorate the expression of NPSLE in MRL/lpr mice. Whether therapeutic depletion or conditional B cell knockout would display similar effects is not yet known.
Autoantibody profiling for Systemic Lupus Erythematosus diagnosis using the Immunarray CHIP™ (P4018)
Authors: Putterman Chaim , Blanco Irene , Jordan Nicole , Daniel-Carmi Vered , Belanis-Meirovich Liron , Sorek Rachel , Cohen-Gindi Ornit , Dromi Nir , Lerner Miriam , Sahar Eli , Batty D. Scott , Tamir Idan , Cohen Irun 
Published: January 2023

Abstract
SLE is a chronic, recurrent, potentially fatal multisystem inflammatory disorder mainly affecting women. SLE patients produce antibodies to many different self-antigens. Yet, currently used serological markers for SLE are lacking in specificity and/or sensitivity. The aim of this study was to develop an improved diagnostic test by measuring and multiplexing specific autoantibody reactivities in SLE patients. Autoantibody reactivity profiles in serum samples collected from 97 SLE patients within three years of the diagnosis were compared with those of 56 healthy controls. Autoantibody profiles were determined using the ImmunArray iCHIP™ - a proprietary protein microarray technology that allows the display of antigens representing a wide range of SLE-associated biochemical pathways on a single chip. Using this novel platform, SLE patients could be differentiated from healthy subjects by a relatively small subset of auto-antigens and Epstein Barr Virus (EBV) antigens. The autoantibody reactivity profile that allowed SLE diagnosis with high sensitivity and specificity displayed differential response to known SLE-specific antigens, such as single-stranded DNA and EBV, and to several novel ones. Conclusion: An antibody profile for SLE diagnosis using a single multiplexed chip was successfully developed. This profile may differentiate between SLE patients and healthy individuals. Validation of this profile in additional patient samples is ongoing.
Modulation of autoimmune diabetes by antibodies specific for N-acetyl-D-glucosamine (P4026)
Authors: New James , Dizon Brian , Kearney John 
Published: January 2023

Abstract
The increasing incidence of autoimmune disease in developed societies has been linked with decreased exposure to environmental antigens, implying that antigen exposure modifies immune system development. Vaccination with Group A Streptococci (GAS) produces a strong antibody response to N-acetyl-D-glucosamine (GlcNAc). This GlcNAc moiety is conserved in mammals, and GlcNAcylated proteins are enriched in pancreatic β-cells. Anti-GlcNAc antibodies generated against GAS bind GlcNAc epitopes in human and murine β-cells. Developmental remodeling of the pancreas is accompanied by significant β-cell apoptosis, which may serve as an initial source of autoantigen priming in Type 1 Diabetes (T1D). We therefore hypothesize that anti-GlcNAc IgM generated during GAS infection mediates non-inflammatory clearance of apoptotic β-cell antigens. Using the Min6 insulinoma cells, we demonstrate that β-cell apoptosis results in surface exposure of GlcNAc residues reactive with anti-GAS Abs. During DC priming with irradiated β-cells anti-GlcNAc Abs suppressed CD4 T cell activation and cytokine production. We show that neonatal immunization with GAS, but not Group C Streptococci, reduces the incidence of diabetes in female NOD-mice. Furthermore, passive transfer of anti-GlcNAc Abs protects NOD.Rag1ko mice from diabetes onset following adoptive transfer of diabetogenic BDC2.5 T-cells. Our data suggest that anti-GlcNAc Abs generated by vaccination with GAS can suppress development of T1D.
The IgM receptor, Toso/FAIM3, is critical for inflammation and diabetes induction. (P4010)
Authors: Lin Gloria Hoi Ying , Marques Ana , Tusche Michael , Brustle Anne , Brenner Dirk , Luk Cynthia , Schroer Stephanie , Winer Daniel , Woo Minna , Mak Tak 
Published: January 2023

Abstract
The mechanisms of type-1 and type-2 diabetes induction are different, yet the two diseases are tightly linked to excessive inflammation. Studies with mice lacking soluble IgM have implicated a protective role of IgM against inflammation and autoimmune progression. Recently, both human and mouse studies have demonstrated Toso/FAIM3, a type one transmembrane protein, to be the receptor for IgM. In our study, we investigated the role of Toso in both type I and type 2 diabetes models in mice. Toso has only been reported to be expressed on B cells and activated T cells. In a type 1 diabetes model, we found that Toso on dendritic cells is required for breaking T cell tolerance and diabetes induction. In a type 2 diabetes model which is induced by 12-14 weeks of high fat diet feeding, WT mice became obese and developed glucose intolerance and insulin resistance. Interestingly, although Toso-deficient mice had similar weight gain, they had improved glucose tolerance and insulin sensitivity compared to the WT mice. Furthermore, Toso-/- mice were found to have decreased B cell number and proinflammatory macrophages in the adipose tissues. In contrast to the protective role of IgM in autoimmune disease progressions, our findings demonstrate a novel and pathogenic role of Toso in both type 1 and type 2 diabetes induction, and suggest that Toso respresents a potential therapeutic target against both autoimmune and type 2 diabetes.
Phospholipid-reactive T cells negatively regulate the function and expansion of glycolipid-reactive T cells (P4012)
Authors: Halder Ramesh , Tran Cynthia , Okorogu Rita , Valera Isela , Prasad Priti , Kim Peter , Singh Ram 
Published: January 2023

Abstract
Glycolipid antigens such as αGalCer and phospholipid (PL) antigens such as phosphatidyl choline bind CD1d and activate T cells. Extensive work on αGalCer-reactive T cells has identified their pathogenic or protective roles in inflammation, infection and cancer. Little is known about the biology and functions of CD1d-restricted PL-reactive T (PL-T) cells. Here, we identified T cells that recognize a battery of PL antigens loaded onto mouse CD1d in various lymphoid organs. While CD1d/PL-T cells were fewer than CD1d/αGalCer-reactive T cells in the liver and spleen, PL-T cells were relatively more abundant than glycolipid-reactive T cells in organs such as mouse decidua. The CD1d/PL-T cells secreted IFN-γ upon in vivo priming with PL antigens in an antigen-specific manner. The PL-T cells did not stain with CD1d/αGalCer tetramers and did not respond to glycolipid antigen αGalCer. Intriguingly, activation of CD1d/PL-T cells markedly reduced the proliferation of αGalCer-reactive T cells in vitro, ex vivo, and in vivo. Thus, PL-T cells are a subset of CD1d-restricted T cells that are phenotypically and functionally distinct from glycolipid-reactive T cells. PL-T cells also negatively regulate the function of glycolipid-reactive T cells. Ongoing studies will investigate whether PL-T cells will modulate the pathogenic and protective roles of glycolipid-reactive T cells in various immune-mediated conditions.
Development of novel Sjogren's Syndrome susceptible mouse lines expressing floxed COX-2 within the major Aec2 disease susceptibility region in chromosome 1 (P4016)
Authors: Mongini Patricia , Haque Shabirul , Chiorazzi Nicholas , Ishikawa Tomo-o , Herschman Harvey , Esposito Donna 
Published: January 2023

Abstract
SS patients exhibit impaired exocrine gland function and high risk for B cell lymphoma. IgG autoAbs to acetylcholine receptors, detectable in humans, impair salivation in SS-afflicted mice with NOD Aec2 and Aec1 susceptibility loci. COX-2 is a potential target for reducing SS pathogenic IgG Ab and B cell lymphoma, since (a) within in vitro cultures B cell-expressed COX-2 and PGE2 augment DNA-modifying AID and downstream IgG switching and (b) mice deficient in COX-2 have reduced IgG. If B cell specific COX-2 function is critical for SS disease, cell-targeted delivery of inhibitors might circumvent adverse side effects. With a downstream goal of examining whether B cell COX-2 contributes to autoimmunity, we introduced a floxed COX-2 gene and a CD19Cre gene independently onto the NOD.B10-H2b SS-susceptible background. To our surprise, early congenic lines HET for either CD19Cre or COX-2flox failed to show aging-related hyposalivation (xerostomia), in contrast to wild-type NOD.B10. Reduced disease in CD19cre HET mice might reflect haplodiploid CD19 or flanking non-NOD genes. Extended backcrossing of COX-2flox onto NOD.B10 yielded two congenic lines that are comparable to wild-type NOD.B10 in developing xerostomia upon aging. Fine genetic mapping of the chr 1 Aec2 region within the novel COX-2flox lines showed that (a) variations in COX-2 do not explain the observed diversity and (b) unidentified genes within a presently more narrowed Aec2 interval are important for xerostomia.