bluuebunny/crossref_metadata_2025_split huggingface dataset:
BANK1 controls IL-6 secretion and a Type I interferon signature via a p38 and MNK1/2- eIF4E translation initiation pathway (P4060)
Authors: Wu Ying-Yu , Kumar Ramesh , Dozmorov Mikhail , Haque Mohammed , Wren Jonathan , Castillejo-López Casimiro , Alarcón-Riquelme Marta 
Published: January 2023

Abstract
BANK1, an adaptor protein expressed in B-cells, is genetically associated with human SLE and plays a little understood role in B-cell signaling. Because BANK1 contains an N-terminal putative Toll-interleukin 1 receptor (TIR) domain, we used murine Bank1-/- splenic B cells to test if BANK1 affects signaling induced by the TLR9 agonist CpG. Following CpG stimulation BANK1 deficiency reduced p38 phosphorylation without affecting that of ERK or JNK and to reduced IL-6 secretion. Bank1-/- cells showed reduced phosphorylation of MNK1/2, confirming that BANK1 affects p38 signaling, and expression microarray analysis showed a disregulated type I interferon response. Microarray analysis and reduced phosphorylation of eIF4E indicated general repression of the eIF4F translation initiation complex in Bank1-/- cells. Together these data establish previously unrecognized roles for BANK1 in CpG-induced responses by splenic B cells. BANK1 may regulate type I IFN responses to TLR9 stimulation by influencing p38 signaling and the eIF4F complex, and thereby influence the course of viral infections or autoimmunity.
Blk haploinsufficiency promotes proinflammatory cytokine production and accelerates onset of kidney disease in B6.lpr mice (P4064)
Authors: Hayes Sandra , Samuelson Elizabeth , Laird Renee , Papillion Amber , Tatum Arthur , Princiotta Michael 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a chronic, multisystem autoimmune disorder whose etiology remains unknown. There is strong evidence for a genetic basis to this disease, and many genetic alleles that predispose an individual to SLE have been identified from studies in patients with SLE and in mouse models of lupus. With recent advances, however, such as the completion of the Human Genome Project and the International HapMap Project, it is possible to perform GWAS to identify additional susceptibility genes in humans. Indeed, several groups using this experimental approach have identified over twenty-five new susceptibility loci in SLE patients of different ethnicity and race. BLK is one of the newly identified susceptibility genes, and risk alleles that map to the BLK locus result in reduced expression. BLK encodes B lymphoid kinase, a Src family kinase whose expression was initially reported to be restricted to the B cell lineage, but we show that it is also expressed in plasmacytoid dendritic cells and IL-17-producing γδ and CD4-CD8- αβ T cells. To investigate the contribution of BLK risk alleles to disease development, we reduced Blk expression in autoimmune-prone B6.lpr mice. We report that Blk+/-.lpr mice exhibit enhanced proinflammatory cytokine production and accelerated onset of lymphoproliferation and kidney disease compared to B6.lpr mice. These data suggest that BLK risk alleles promote disease through the dysregulation of a proinflammatory cytokine network.
Differential regulation of immune responses in MyD88-deficient mice by cytokine-expressing Salmonella typhimurium (P4046)
Authors: al-Ramadi Basel , Issac Jincy , El-Ojali Samia , Mohamed Yassir , Al-Ketbi Mai , Fernandez-Cabezudo Maria 
Published: January 2023

Abstract
MyD88 is a critical component of most TLR signaling pathways. Earlier we demonstrated that MyD88-deficient (MyD88-/-) mice are hypersusceptible to infection by a vaccine strain (aroA-/aroD-) of Salmonella typhimurium (designated BRD509). Curiously, infected mice exhibited a state of hypergammaglobulinemia, with high levels of anti-Salmonella Abs of all IgG isotypes. The dysregulated Ab response also lead to production of anti-dsDNA and anti-thyroglobulin autoantibodies and immune complex deposition in kidneys. Increased susceptibility of MyD88-/- mice has been linked to poor type-1 cytokine responses. Therefore, we assessed the response of these mice to infection with a S. typhimurium engineered to express murine IFN-γ (GIDIFN). Infection studies using different doses of BRD509 and GIDIFN strains revealed that the latter strain was &gt;100-fold less virulent in MyD88-/- mice than BRD509 and correlated with decreased bacterial loads in systemic organs and robust pathogen clearance starting at 7 days of infection. Although GIDIFN-infected mice exhibited high Ab titers, the degree of autoreactivity was significantly decreased compared to BRD509-infected animals. Our data suggest that TLR-MyD88 signaling pathway is critical for controlling Ab responses to bacterial antigens and for maintaining B cell self-tolerance and demonstrate that the two processes may be unlinked through the use of cytokine-expressing bacterial strains that modulate host immune responses.
A dominant CD4+ T cell response to Helicobacter pylori is associated with decreased risk of gastric diseases in human (P4328)
Authors: Zou Quanming 
Published: January 2023

Abstract
BACKGROUND and AIMS: Immunodominance has been well-demonstrated as a key feature in antiviral, antitumoral and antibacterial cellular immune responses but much less so in the immune responses against Helicobacter pylori (H. pylori). Antigen-specific CD4+ T cells have been reported to play a pivotal role in protecting against H. pylori in murine systems. We investigated the immunodominant CD4+ T cell response to HpaA (also called neuraminyllactose-binding hemagglutinin), a conserved, H. pylori-specific colonization factor which has been considered as a promising vaccine candidate. METHODS: HpaA-specific CD4+ T cells were expanded with autologous peripheral blood mononuclear cells (PBMCs) loaded with recombinant HpaA and further characterized with overlapping synthetic peptides. The percentage of HpaA88-100-specific CD4+ T cells restricted to HLA-DRB1*1501 from 59 H. pylori-infected subjects with different diseases were compared. RESULTS: Several novel, immunodominant CD4+ T cell epitopes were identified and characterized. The immunodominant CD4+ T cell responses specific to HpaA88-100 were observed in most of the H. pylori-infected individuals who expressed HLA-DRB1*1501 and they were significantly more abundant in patients with less severe diseases (P&lt;0.05). CONCLUSIONS: The HLA-DRB1*1501-restricted immunodominant CD4+ T cell response specific to HpaA88-100 is associated with decreased risk of severe gastric diseases. Further study of these and other similar immunodominant CD4+ T cell responses to H. pylori will help the field to gain critical insight into the mechanism of protective immunity against H. pylori in humans and the knowledge gained will improve potential vaccine design.
Tracking the CD4+ T cell response to BCR-ABL-induced leukemia (P4330)
Authors: Berquam-Vrieze Katherine , Farrar Michael 
Published: January 2023

Abstract
Patients with BCR-ABL+ acute lymphoblastic leukemia develop T cell responses to the BCR-ABL fusion peptide (referred to as BAp). However, these responses are not sustained and loss of the BAp-specific T cell response correlates with relapse. A major gap in our knowledge is why such epitopes do not elicit immune responses that eliminate BCR-ABL+ leukemia. This knowledge gap exists because of an inability to track the fates of endogenous BAp:I-Ab-specific T cells in vivo. To address this question we plan to generate peptide:MHCII tetramers that will allow us to track BAp:I-Ab-specific T cell responses. We have demonstrated that CD4+ T cells exist that recognize the BCR-ABL fusion junction in a mouse model of BCR-ABL+ leukemia. We are generating BAp:I-Ab tetramers that will allow us to track these cells in vivo. Using this reagent we will determine whether T cell responses are impaired due to deletion, immune deviation, Treg-mediated suppression or anergy. We will then assess whether enhancing antigen presentation or inhibiting anergy improves BAp:I-Ab-specific T cell responses. In addition, we will determine if altering the cytokine mileu can improve the CD4+ T cell response to BCR-ABL+ leukemia using both knock-out mouse mice and in vivo delivery of exogenous cytokines.
Use of mRNA for vaccination or gene therapy: delivery route determines the verdict (P4316)
Authors: Pollard Charlotte , Vandermeulen Gaëlle , Vanham Guido , Grooten Johan , Préat Véronique , De Koker Stefaan 
Published: January 2023

Abstract
The use of DNA and viral vector vaccines for the induction of cellular immune responses is increasingly gaining interest. However, concerns have been raised regarding the safety of these immunization strategies. Due to their lack of genome integration, mRNA-based vaccines have emerged as a promising alternative. In this study we evaluated the potency of antigen-encoding mRNA either delivered via complexation with the cationic lipid DOTAP/DOPE, or delivered via in vivo electroporation as a novel vaccination approach. By using luciferase-encoding mRNA we showed that in vivo electroporation of mRNA induces high levels of luciferase at the injection site, while protein expression in mice injected with DOTAP/DOPE complexed mRNA was barely above background levels. Despite the high levels of protein expression, in vivo electroporation of mRNA failed to elicit immune responses against the encoded antigen. In contrast, lipid-based delivery of mRNA induced antigen-specific, functional T cell responses resulting in specific killing of peptide-pulsed cells and the induction of humoral responses. In summary, in vivo electroporation of antigen-encoding mRNA induces high levels of protein expression but no immune responses, thus presenting an ideal tool for mRNA-based gene therapy, while lipid-based delivery offers advantages for vaccination purposes due to the induction of low protein levels but strong antigen-specific cellular immunity.
An effective cell-based approach for the generation and identification of functional monoclonal antibodies to cell surface targets (P4320)
Authors: Zhang Chonghui , Chan Nancy , Kikuchi Lena , Voegeli Karin 
Published: January 2023

Abstract
Monoclonal antibodies (mAbs) have long been remarkable tool reagents for discovery research, but have also recently emerged as a significant class of therapeutic biologics. The development of therapeutic mAbs, however, has been hindered by the challenges of purifying distinct antigens like cell membrane-bound proteins and by the lack of effective methods for identifying mAbs that recognize native epitopes of the antigen. Here we developed an effective cell-based approach for generating mAbs against cell surface targets. Targeted antigens of interest were molecularly cloned and stably expressed on the cell surface of a mouse pre-B cell line. The transfected cells were then used for immunization of a syngeneic strain of mice and for subsequent hybridoma selection by high-throughput flow cytometry. Panels of mAbs were raised against a large number of antigens ranging from cell surface proteins to viral targets. The mAbs generated by this approach were predominantly IgG isotypes and had minimal binding affinities of a nanomolar range. Most of the mAbs recognized antigens in their native conformation and were diversified in their Ig sequences. More importantly, many of these mAbs were found to have functional activity, such as receptor antagonism, protein kinase inhibition or neutralizing activity. This cell-based approach for immunization and mAb selection offers an effective means to generate high-quality mAbs for the needs of discovery research and therapeutic development.
Rewiring of the Bcr/Abl and PKCβII signal transduction pathways (P4322)
Authors: Hoekstra David , Carlson Louise , Lee Kelvin 
Published: January 2023

Abstract
The ability to differentiate leukemia cells directly into antigen presenting cells (APCs) in vivo has long been sought after as a method of cancer immunotherapy. While also being less proliferative, these newly differentiated APCs may present new antigens to the immune system and could potentiate a robust immune response. We have previously shown that canonical activation of protein kinase C isoform βII (PKCβII) drives dendritic cell differentiation and apoptosis in leukemic cells. With this as our pro-drug, we have rewired its signaling to be activated by the oncogenic kinase Bcr/Abl, which is essential for chronic myeloid leukemia cell survival. To do this, the abl kinase target motif was inserted into a specific regulatory sequence in PKCβII based off of molecular modeling techniques. Using ImageStream technology, we have found that the rewired PKCβII constructs are activated in a Bcr/Abl+ leukemia cell line, but not in Bcr/Abl- cells; introduction of Bcr/Abl into non-expressing cells allows for activation of the rewired PKCβII. Additionally, this activation was blocked by the Bcr/Abl inhibitor imatinib in leukemia cells sensitive to imatinib but persisted in resistant cells. Activation of PKCβII by Bcr/Abl led to a decrease in viability and an increase in apoptosis in Bcr/Abl+ leukemic cells. This novel strategy of activating differentiation machinery in leukemic cells in vivo provides a new method of treating patients who do not respond to traditional chemotherapy.
mTORC1 inhibition blocks the gut-homing phenotype induced by retinoic acid in naturally-occurring Treg but not in conventional CD4+ T cells (P6260)
Authors: Chen Leo , Rosborough Brian , Thomson Angus , Raimondi Giorgio 
Published: January 2023

Abstract
The vitamin A metabolite all-trans retinoic acid (ATRA) is known to induce a gut-homing phenotype in T cells by upregulating the integrin α4β7 and CCR9. We report that, unlike conventional CD4+ T cells, only about half of naturally-occurring Treg (nTreg) upregulate CCR9 in response to ATRA. The bimodal distribution of CCR9+ versus CCR9- Treg subsets was not due to differences in the extent of proliferation, intensity level of FoxP3, or expression of the nTreg/iTreg discriminator Helios. Furthermore, we found that addition of the mammalian target of rapamycin (mTOR) inhibitor rapamycin prevented the appearance of the CCR9+ subset of nTreg, but did not prevent CCR9 upregulation by T cells, suggesting nTreg-specific involvement of mTOR in regulation of the expression of CCR9. mTOR involvement was independent of Akt activity, as inhibition of Akt or its downstream target GSK-3β did not prevent CCR9 expression. Additionally, Rictor-/- Treg (which lack mTORC2) showed no diminution of the CCR9+ subset compared to wild-type controls, demonstrating selective participation of mTORC1. These results suggest that nTreg are a heterogeneous population with two subsets: one that does not upregulate CCR9 in response to ATRA, and one that does upregulate CCR9 in an mTORC1-dependent fashion. These findings reveal a novel difference between nTreg and conventional T cells, and provide a framework for which the migratory behavior of nTreg versus T cells may be differentially influenced.
Effects of osteopontin on B cells (P6285)
Authors: Dsilva Sehba , Rothstein Thomas 
Published: January 2023

Abstract
Osteopontin (OPN) is a protein responsible for diverse processes. OPN expression is linked to autoimmunity and cancer. Both innate and adaptive immune cells like macrophages, dendritic cells,T and B cells express OPN. OPN has many effects on non-B immune cells, the source of these effects is either the OPN produced by the cells themselves or OPN made by other cells. With regard to B cells, it’s been suggested that high levels of OPN contribute to chronic and sustained polyclonal B cell activation. However, the specific effects of OPN on B cell survival, migration and stimulated proliferation are unknown. Our objective is to identify these specific effects of OPN on B cells and determine the contribution to disease activity. For survival studies, purified B cells from WT mice-untreated and treated with exogenous OPN were used along with B cells from OPN transgenic mice. Increased survival was seen in both OPN treated B cells and B cells from transgenic mice. Thymidine incorporation and CFSE assays were used to examine proliferation of B cells from WT and OPN transgenic mice in response to various stimuli. The results indicate enhanced proliferation in B cells from the transgenic mice compared to those from WT mice. Using transwell migration assay, preliminary experiments also indicate that OPN induces migration of B cells. These developments suggest that OPN affects B cell survival, proliferation and migration, which could in turn contribute to propagation of disease activity.
The RNA-binding protein HuR coordinately regulates GATA-3 and Th2 cytokine gene expression in dose dependent manner (P6262)
Authors: Atasoy Ulus , Gubin Matthew , Calaluce Robert , Magee Joseph , Martindale Jennifer , Gorospe Myriam , Casolaro Vincenzo , Stellato Cristiana 
Published: January 2023

Abstract
Posttranscriptional control by RNA binding proteins (RBPs) is poorly understood. Th2 mediated diseases such as asthma, are driven by GATA-3, IL-4 and IL-13. The RBP, HuR, regulates IL-4 and IL-13 by increasing mRNA stability. We identified GATA-3, IL-4 and IL-13 as HuR targets and hypothesized HuR may be coordinately regulating Th2 differentiation by controlling mRNA stability and translatability. We made a HuR CD4+T transgenic mouse, and a HuR conditional knock out mouse (HuRfl/fl) to ablate HuR in T cells. HuR over-expression in transgenic CD4+ T cells stabilized GATA-3, IL-4 and IL-13 mRNAs, and increased mRNA, protein and Th2 polarization. HuR knock-down resulted in decreases in GATA-3, IL-4 and IL-13 expression. Findings were confirmed in human naïve and memory T cells. We defined HuR binding sites in GATA-3. Th2 cells with low HuR levels (26%) from HuRfl/+ mice, had decreases in IL-4, IL-13 and GATA-3 mRNA but not protein. Surprisingly, CD4+ T cells from HuRfl/fl mice (93% knockdown) showed increased IL-2, IL-4, IL-13 mRNA and protein. IL-4 transcription increased but mRNA stability remained unchanged. IL-2 and IL-13 transcription were unaltered but there were increases in mRNA stability. Polysomal gradient analysis revealed similar translation. We analyzed gene expression in KO vs. control CD4+ T cells to determine HuR targets. Ova challenge model of asthma corroborated ex vivo results. These data suggest HuR levels regulate Th2 cytokines and Th2 differentiation.
The TL1A-IL13 axis: a novel pathway for Type 2 immunity mediated by innate lymphocytes independent of helminth infection (P6264)
Authors: Meylan Francoise , Hawley Eric , Barron Luke , Barlow Jillian , Penumetcha Pallavi , Richard Arianne , Chen Xi , Paul William , Wynn Thomas , McKenzie Andrew , Siegel Richard 
Published: January 2023

Abstract
The TNF-family cytokine TL1A costimulates T cells through its receptor DR3 and is required for diverse autoimmune disease models, but its role in innate lymphocyte biology has not been explored. Transgenic mice chronically expressing TL1A have increased activated T cells and develop small intestinal pathology characterized by high levels of IL-13, muscular and goblet cell hyperplasia, and infiltration with immune cells, all of which depend on DR3. Genetically eliminating T cells reduces T cell hyperactivation and gut infiltration with lymphocytes but not IL-13 production or mucosal hyperplasia. These data suggest that TL1A can induce another cell type to produce IL-13 and induce gastrointestinal pathology. Using a DsRED reporter mouse for IL-13, we find that the major IL-13 producing cells in TL1A transgenic mice lack T and B cell lineage markers and are phenotypically similar to ’type 2’ innate lymphocytes (ILC2) which expand and produce IL-13 in response to IL-25, IL-33 or parasitic infection. TL1A directly promotes IL-13 production by ILC2 ex-vivo. However, DR3 deficient mice mount a vigorous response to the intestinal nematode Nippostrongylus brasiliensis, and IL-13 producing innate lymphocytes can be expanded by IL-25 in the absence of DR3. Thus, TL1A can directly induce IL-13 production by innate lymphocytes through mechanisms distinct from parasitic infection. TL1A may be an attractive target for immunotherapy of diverse diseases associated with this cytokine.
Interleukin 32 promotes angiogenesis: a new role for a versatile cytokine. (P6266)
Authors: Rudloff Ina , Zepp Jarod , Dinkel Hannah , Palmer Brent , Farkas Laszlo , Cool Carlyne , Taraseviciene-Stewart Laima , Kim Soo-Hyun , Dinarello Charles , Voelkel Norbert , Nold Claudia , Nold Marcel 
Published: January 2023

Abstract
IL-32 is a multi-faceted cytokine with roles in infection, autoimmunity and cancer. It aggravates inflammation, inhibits virus propagation and acts as a critical regulator of endothelial cell (EC) function. We now discovered that IL-32 also possesses angiogenic properties. The hyperproliferative EC of pulmonary hypertension and glioblastoma multiforme exhibited a markedly increased abundance of IL-32, and the cytokine colocalized with integrin αVβ3. In vitro, VEGF receptor blockade resulted in hyperproliferation of EC and a 3.4-fold increase in IL-32. When IL-32 was silenced by siRNA, the proliferation of adult and fetal EC was reduced (change in cell number +58% vs -15%). Mechanistic studies revealed that siIL-32-transfected EC produced 3-fold more activin A and angiopoietin 2, but 61% less NO, whereas neither apoptosis nor the abundance of VEGF or TGF-β were affected. Interestingly, pre-treatment with IFNγ was required to render EC responsive to exogenous IL-32γ. To obtain in vivo evidence for these angiogenic effects, we injected mice with matrigel plugs loaded with IL-32γ, VEGF or vehicle, and performed CD31- and H&E-staining after 14d. As anticipated, 25 ng/ml of VEGF increased neocapillarization 8-fold, but unexpectedly the effect of 100 ng/ml IL-32γ was even stronger (8.5-fold). In summary, we add VEGF-independent angiogenic properties to the portfolio of IL-32, implicating a role in pulmonary hypertension and neoplastic diseases that is possibly exerted via αVβ3.
Dendritic cell derived exosomes need to activate both T and B cells to induce antitumor immunity (P4250)
Authors: Näslund Tanja , Gehrmann Ulf , Qazi Khaleda , Karlsson Mikael , Gabrielsson Susanne 
Published: January 2023

Abstract
Exosomes are secreted membrane nanovesicles of endosomal origin and are considered as potential cancer vaccine vectors. Phase I clinical trials have been successfully conducted with tumor peptide-loaded exosomes derived from dendritic cells (dexosomes) and a phase II clinical trial is currently ongoing. However, much is still unknown regarding the in vivo role of dexosomes and if the immunogenicity of dexosomes can be enhanced. We have previously reported that dexosomes induce CD4+ T cell responses in a B cell dependent manner, suggesting that immunizing with dexosomes only carrying T cell peptides induce suboptimal immune responses. Here we show that CD8+ T cell responses were induced in vivo when mice were immunized with protein, but not peptide, loaded dexosomes. We could also show that the cytotoxic T cell response was totally dependent on CD4+ T cells and, interestingly, also on B cells. Mice deficient in complement activation and antigen shuttling by B cells have lower responses to protein-loaded dexosomes showing involvement of these B cell mediated mechanisms. Finally, protein-loaded dexosomes were superior in protecting against tumor growth. In conclusion, proper activation of CD4+ T and B cells need to be considered when designing cancer vaccines to ensure full potential of the treatment.