bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel small molecule inhibitor targeting gp130 (CCR3P.217)
Authors: Heo Tae-Hwe , Park Kyung-Yeon , Park Yeon-Hwa 
Published: January 2023

Abstract
Interleukin-6 (IL-6) is a major causative factor of inflammatory disease. Although IL-6 and its signaling pathways are promising targets, orally available small molecule drugs specific for IL-6 have not been developed. To discover IL-6 antagonists we screened our in-house chemical library and identified a novel synthetic compound as a candidate IL-6 blocker. The activity, mechanism of action, and direct molecular targets of the compound were investigated. A reporter gene assay showed that the compound suppressed activation of signal transducer and activator of transcription-3 (STAT3) induced by IL-6, but not activation induced by leukemia inhibitory factor. In addition, the compound downregulated IL-6-stimulated phosphorylation of STAT3, gp130, and JAK2 protein in HepG2 cells and substantially inhibited IL-6-dependent TF-1 cell proliferation. The compound antagonized IL-6-induced TNF-α production in vivo, but did not affect IL-2-induced TNF-α production. In pathologic models, oral administration of the compound ameliorated collagen-induced arthritis and acute pancreatitis in mice. We subsequently demonstrated direct and specific interaction between the compound and gp130, which was measured by SPR analysis (KD = 7.4 μM) and was simulated by a docking study. Taken together, our data suggest that the compound is a novel synthetic IL-6 inhibitor that functions through direct binding to gp130.
Identifying novel mechanisms regulating expression of inflammatory chemokine CXCL2 (CCR3P.206)
Authors: Zhang Xian , Lou Ning , Lennard Richard Mara , Yu Jin , Kindy Mark 
Published: January 2023

Abstract
Mammalian cells produce inflammatory mediators in response to the innate immune signal and their expression is tightly regulated. Chemokine (C-X-C motif) ligand 2 (CXCL2), also known as MIP2-alpha, belongs to the CXC chemokine family. CXCL2 is chemotactic for neutrophils. Elevated expression of CXCL2 is associated with many inflammatory and autoimmune diseases. Recently we have found that the Fli-1 transcription factor is a novel regulator in modulating the expression of many inflammatory mediators. including MCP-1,CCL5) and IL-6. Since we found that lupus-prone mice with reduced expression of Fli-1 had significantly decreased neutrophil infiltration into the kidneys, we investigated if Fli-1 regulates the expression of CXCL2. We found that Fli-1 protein expression in endothelial cells transfected with Fli-1 specific siRNA was significantly decreased compared to the expression of Fli-1 in cells transfected with control siRNA after stimulation by Toll-like receptor (TLR) 4 ligands, and tumor necrosis factor (TNF) alpha. The production of CXCL2 in endothelial cells with LPS stimulation is dose-dependent. We found that Fli-1 binds to the CXCL2 promoter by Chromatin immunoprecipitation‎ (ChIP) assay. Transient transfection assays show that Fli-1 drives transcription from the CXCL2 promoter in a dose-dependent manner. These results indicate that Fli-1 is a novel, critical transcription factor in regulating the expression of the inflammatory chemokine CXCL2.
Interleukin-12 elicits various responses of splenocytes from different mouse strains (CCR3P.207)
Authors: Nguyen Khue , Koppolu Bhanu , Smith Sean , Ravindranathan Sruthi , Zaharoff David 
Published: January 2023

Abstract
Interleukin - 12 (IL-12) is a pivotal linkage between innate and adaptive immunity via its pleiotropic functions on immune system. IL-12 has been shown to provide anti-tumor signals in mouse models including the ability to recruit immune cells, induce the production of interferon - γ (IFN-γ), tumor necrosis factor (TNF) and promote anti-angiogenesis. IL-12 bioactivity significantly varies between mouse strains. However, the mechanism resulting in various responses remains unclear. In this research, bioactivity of IL-12 was determined by quantifying IFN- γ production by splenocytes collected by three different mouse strains (C3H/HeJ, C57BL/6J, BALB/cByJ) in response to IL-12. Our data show that C3H/HeJ mouse strain produces increased levels of IFN-γ when compared with other mouse strains. IL-12 receptor (IL-12RB1) of splenocytes was quantified by fluorescence-activated cell sorting (FACS). Our studies show that the expression of IL-12RB1 by several key immune cell types including CD4 T-cells, CD8 T-cells, NK T-cells and B cells was significantly higher in C3H/HeJ splenocytes compared to splenocytes from C57BL/6J or BALB/cByJ. The higher expression of IL-12RB1 by C3H/HeJ mouse splenocytes leads to a significantly stronger response to IL-12 compared to other mouse strains. Our studies provide a valuable insight into variation of IL-12 bioactivity between different individuals. Further studies will be conducted to better understand in vivo bioactivities of IL-12 on mouse models.
The role of miR-204 in early B cell development. (HEM1P.219)
Authors: Medina Kay , Xu Zhiuhu , Gwin Kimberly 
Published: January 2023

Abstract
The generation of B lineage lymphocytes in bone marrow is contingent upon the combinatorial activities of transcription factors, signaling molecules, chromatin modifiers, and microRNAs. One transcription factor important for the generation of B cell precursors is the homeodomain protein HoxA9. Equally important as HoxA9 function in facilitating lymphoid lineage specification is silencing HoxA9 activity upon commitment to the B cell fate as sustained expression impairs later stage B cell differentiation. We previously showed that transcripts for the B cell specification factor EBF1 show an inverse expression pattern to HoxA9. Furthermore, EBF1 does not directly suppress HoxA9 transcription. Hox proteins are regulated by microRNAs in some developmental contexts. MicroRNA profiling of an EBF1-/- cell line stably expressing an inducible EBF1:ER fusion protein revealed a single miR induced by EBF1 activity, miR-204. Importantly, forced expression of miR-204 in EBF1-/- cells repressed transcription and protein levels of the HoxA9 co-factor, Meis1. We hypothesized that if Meis1 is a critical target of miR-204, then sustained expression of miR-204 should phenocopy conditional deletion of Meis1 or HoxA9. Indeed, we determined that forced expression of miR-204 in wildtype multipotent progenitors impaired B cell differentiation, similar to loss of HoxA9. These data suggest a novel role for EBF1 in B cell development, induction of miR-204 to silence a HoxA9-Meis1 transcriptional axis.
c-Myb coordinates survival and expression of key signaling components required for the pre-BCR checkpoint (HEM1P.220)
Authors: Bender Timothy , Daamen Andrea , Fahl Shawn 
Published: January 2023

Abstract
The c-Myb transcription factor is abundantly expressed during B cell development and is down regulated as the cells mature. We targeted Myb in mice with loxP sites (Mybf/f). By crossing these mice to other strains that direct Cre expression to different stages of B cell development (Rag1-cre, Mb1-cre, CD19-cre) we are able to assess subsequent B-lineage maturation. In Mybf/f Mb1-cre mice, c-Myb is required for B cell differentiation beyond the pre-pro-B cell stage and survival of CD19+ pro-B cells. c-Myb controls survival of pro-B cells by suppression of the proapoptotic factors Bmf and Bim. We find that IL-7 signaling represses the expression of Bmf and Bim. c-Myb controls the expression of CD127 but forced expression of CD127 is not sufficient to repress Bmf and Bim or rescue survival of c-Myb deficient pro-B cells. Examination of negative mediators of IL-7 signaling revealed increased SOCS3 mRNA in c-Myb deficient pro-B cells and forced expression of SOCS3 suppressed proliferation and survival of CD19+ pro-B cells in the presence of IL-7. Surprisingly, Mybf/f CD19-cre Bcl2-Tg mice contain severely reduced large and small pre-B cell compartments. Examination of mRNA expression in these cells revealed that c-Myb controls expression of key regulators of the pre-BCR checkpoint, including CD127, lambda-5, cyclin D3 and Cxcr4. c-Myb coordinates survival in pro-B cells with the expression of key components of the signaling pathways that drive the pro-B to pre-B cell transition.
Abnormal thymocyte development in Mbnl1 knockout mice (HEM2P.240)
Authors: Scotti Marina , Shin Jihae , Batra Ron , Finn Dustin , Swanson Maurice 
Published: January 2023

Abstract
Transcriptional regulation has been shown to play a vital role in thymocyte development, however co-/post-transcriptional modifications are also important regulatory mechanisms for T cell development. Here, we demonstrate that loss of Mbnl1, a developmentally regulated alternative RNA processing factor, leads to dysregulation of thymocyte development. Mbnl1-/- mice develop thymic hyperplasia due to accumulation of CD4+CD8+ double positive thymocytes. Mbnl1-/- thymocytes are resistant to steroid-induced apoptosis in vitro and current studies are focused on characterizing Mbnl1-/- thymocyte proliferation defects as well as peripheral T cells in Mbnl1-/- mice. Whole thymic transcriptome comparisons between Mbnl1-/- and wild type mice indicate 476 mis-splicing events in &gt;300 genes, including those involved in T cell receptor assembly and signaling. Additionally, 688 differentially expressed genes identified between wild type and Mbnl1-/- (FDR &lt; 0.01) demonstrate enrichment for immune processes, such as lymphocyte activation and stimulus response. Since Mbnl1 regulates alternative splicing and polyadenylation during the postnatal period, this study suggests that fetal to adult RNA processing switches are critical for normal thymocyte development.
Divergence in the requirements for HEB factors in the generation and function of fetal-derived versus adult-derived γδ T cells (HEM2P.245)
Authors: Anderson Michele , In Tracy 
Published: January 2023

Abstract
Gamma-delta (γδ) T cells are involved in mucosal immunity, wound healing, and tumor immunosurveillance. γδ T cells are programmed in the thymus to produce either IL-17 or IFNγ, but the molecular basis of this programming is not well understood. Here we show that mice lacking the E protein transcription factor HEB are defective in their ability to generate Vγ4 γδ T cells in fetal thymic organ culture. Since the HEB null allele is embryonic lethal, we generated mice lacking HEB only in hematopoietic cells by crossing HEBfl/fl mice with Vav-Cre mice (HEBfl/fl;vav-cre). These mice show a partial block in αβ T cell development, as seen in the fetal thymus of HEB-/- mice. In stark contrast to fetal gd T cells, however, the frequency of Vγ4 γδ T cells in the thymus of the HEBfl/fl;vav-cre mice is the equivalent to that in the WT thymus. Nonetheless, there is a skewing toward the CD27+ γδ T cells and away from CCR6+ γδ T cells, in both Vγ4+ and Vγ4- populations, suggesting a biased programming towards an IFNγ producing fate and away from the IL-17 producing fate. Moreover, there is a selective expansion of CD27+ Vγ4+ γδ T cells in the spleen, at the expense of other γδ T cell subsets, and a loss of CCR6+ Vγ4 γδ T cells in the lungs of HEBfl/fl;vav-cre mice. These results reveal that fetal Vγ4 cells are HEB-dependent, while adult Vγ4 cells appear to not require HEB, and strongly suggests that HEB is required for the generation and/or maintenance of IL-17 producing γδ T cells.
HIV-1 promotes neutrophil infiltration and lung damage in humanized mice co-infected with Mycobacterium tuberculosis (HUM1P.266)
Authors: Nusbaum Rebecca , Huante Matthew , Sutjita Putri , Calderon Veronica , Vijayakumar Sudhamathi , Aronson Judith , Hunter Robert , Actor Jeffrey , Cirillo Jeffrey , Valbuena Gustavo , Endsley Janice 
Published: January 2023

Abstract
Co-infection with human immunodeficiency virus (HIV) and Mycobacterium tuberculosis (Mtb) promotes aggressive disease through mechanisms that are poorly understood. A recent study suggests an association between peripheral blood neutrophil count and Mtb load in the sputum of human subjects with HIV co-infection. We used our humanized BLT mouse (HuMouse) model of HIV/Mtb co-infection as an investigative tool to study the role of neutrophils in co-infection pathobiology. Humanized mice were infected intravenously with HIV-1 for 3 weeks and then co-infected intranasally with Mtb. Compared to HuMice with Mtb or HIV mono-infections, increased neutrophil numbers were observed in the lungs of co-infected animals, as determined by detection of myeloperoxidase using immunohistochemistry. Consistent with the role of neutrophils to promote immune-mediated pathology in TB disease, greater tissue necrosis and increased mycobacterial burden was observed in the lungs of co-infected animals. In support of these findings, an increased pulmonary production of cytokines and chemokines associated with neutrophil recruitment including IL-8, CXCL5, fractalkine, and IL-17 were also observed in co-infected animals. These studies identify mechanisms for immune-mediated pathology due to HIV/Mtb co-infection that can be targeted to restore immune balance in the lung and complement antimycobacterials.
Oxidized low density lipoprotein immune complexes act as a priming signal for the NLRP3 inflammasome (HUM1P.253)
Authors: Rhoads Jillian , Wilhelm Ashley , Moore Jared , Major Amy 
Published: January 2023

Abstract
Antibodies to oxidized LDL (oxLDL) and resulting immune complexes (ICs) are a prominent feature of atherosclerosis and autoimmune diseases associated with increased atherosclerosis. Although levels of oxLDL-ICs correlate with disease severity, it is unclear whether these ICs play an active role in disease pathogenesis. We hypothesize that oxLDL-ICs exacerbate atherosclerosis by signaling through multiple receptors on dendritic cells (DCs) resulting in an inflammatory adaptive immune response. To test this hypothesis, bone marrow derived dendritic cells (BMDC) were incubated with oxLDL or oxLDL-IC and cytokine levels in culture supernatants were measured by ELISA. Results indicated a 10-fold increase in IL-1β production from BMDC treated with oxLDL-IC compared to oxLDL, suggesting a role for the inflammasome in oxLDL-IC activation of DC. Pre-treatment of BMDC with a caspase 1 inhibitor and use of NLRP3-/- BMDC abolished IL-1β secretion in response to oxLDL-IC, further implicating the inflammasome. Additional investigation using Fab fragments and a TLR4 inhibitor uncovered a role for both FcγR and TLR4 in oxLDL-IC mediated IL-1β production. Finally, BMDC pre-treated with oxLDL-IC, but not oxLDL, and incubated with CD4+ OT-II T cells in the presence of OVA peptide skewed T cells towards a Th17 phenotype. Taken together, these data suggest a novel and pathologic role of oxLDL-ICs in activating the NLRP3 inflammasome and promoting an inflammatory T cell phenotype.
Hyper-responsive TLR7 and TLR9 activation in NADPH oxidase-deficient human B cells (HUM1P.272)
Authors: McLetchie Shawna , Gardiner Gail , Blum Janice 
Published: January 2023

Abstract
Chronic Granulomatous Disease (CGD) is an inherited immunodeficiency linked with mutations in the multi-component leukocyte NADPH oxidase. Phagocytic cells deficient in NADPH oxidase fail to produce sufficient levels of the reactive oxygen species (ROS) necessary to clear engulfed pathogens. Though the hyperactive immune response associated with CGD is widely described in phagocytes, the effects of oxidase-deficiency on B cell functions has not been well studied. In B cells, recognition of microbial single-stranded RNA or unmethylated DNA by intracellular toll-like receptors (TLR) 7 and 9, respectively leads to NADPH oxidase derived ROS production. Here, studies demonstrate hyperactive responses to endosomal TLR stimulation in B cell lines derived from patients with mutations in either the p40phox or p47phox subunits of the NADPH oxidase which compromise its function. NADPH oxidase-deficient B cell lines expressed enhanced levels of TLR7 and TLR9 mRNA compared to cells reconstituted to restore oxidase activity. TLR7/9 stimulation of oxidase-deficient B cells resulted in greater cytokine production and p38 MAP kinase activation than oxidase-sufficient B cells. These data are consistent with the autoimmune predisposition associated with CGD and suggest that B cells may also contribute to hyperactive immune responses detected in individuals with CGD.
The role of indoxyl sulfate, a circulating uremic toxin, for inflammatory responses in the patients with end-stage renal disease (HUM1P.304)
Authors: Hwang Yuri , Jang Ji yeon , Chung Yeon-Ho , Kim Bonah , Kim Dong Hyun , Park Sungha , Chung Doo Hyun , Lee Won-Woo 
Published: January 2023

Abstract
In the ESRD patients, a major cause of death is cardiovascular disease (CVD) and its pathologic processes have been suggested to link to uremia-related chronic inflammation. Although concept of ESRD-related immune dysfunction is well-accepted, little is known about how uremic toxins affect cellular immunity involved with pathogenesis of CVD in the patients. Thus, we investigated phenotypic and functional features of CD4 T cells and monocytes in the ESRD patients and their immune responses mediated by indoxyl sulfate (IS), a key uremic toxin in order to explore the pathogenic roles of these cells for vascular endothelial cells (VEC). In ESRD patients, CD4+CD28null T cells and CD16+ monocytes were expanded as compared with HC. To explore how uremic milieu affects immune responses, monocytes were stimulated with IS. These monocytes produced a large amount of TNF-α through aryl hydrocarbon receptor. TNF-α stimulated VEC greatly produced CX3CL1, a ligand of CX3CR1 which is overexpressed by CD4+CD28null T cells and CD16+ monocytes. CD4+CD28null T cells are preferentially recruited by CX3CL1 and moreover, CD4+CD28null T cells have cytotoxic capability allowing for induced apoptosis of VEC in response to TCR stimulation. Our findings suggest that IS-mediated immune dysfunction may play a critical role for development and accelerated progression of CVD through VEC damage in the ESRD patients.
The MTA1-TG2 is involved in self-limitation of monosodium urate crystal-induced inflammation by upregulating the levels of active TGFβ1 leading to inhibition of JAK2 signaling (HUM1P.305)
Authors: Tsay Gregory , Yen Jia-Hau , Lin Ling Chung , Leong Pui Ying , Chen Jiunn-Horng , Szondy Zsuzsa 
Published: January 2023

Abstract
Abstract Introduction: Transglutaminase 2 (TG2), a protein crosslinking enzyme with multiple biochemical functions, has been connected to various inflammatory processes. In this study, the involvement of TG2 in monosodium urate (MSU) crystal-induced inflammation was studied. Conclusions: These findings reveal an inherent regulatory role of the MTA1-TG2 pathway in the self-limitation of MSU crystal-induced inflammation via positively regulating the levels of active TGF-b in macrophages that opposes the MSU crystal- induced JAK2 -dependent pro-inflammatory cytokine formation.
Loss of NLRX1 results in increased intestinal pathology and T cell responses in mice with inflammatory bowel disease (HUM1P.312)
Authors: Eden Kristin , Hontecillas Raquel , Viladomiu Monica , Washington Philipson Casandra , Carbo Adria , Leber Andrew , Philipson Noah , Tattoli Ivan , Girardin Stephen , Allen Irving , Bassaganya-Riera Josep 
Published: January 2023

Abstract
NLRX1 is a mitochondrial-associated NOD-like receptor that modulates antiviral immunity, cellular stress, autophagy, and reactive oxygen species (ROS). The role of NLRX1 in inflammatory bowel disease remains unknown. This study aimed to characterize NLRX1-mediated mechanisms of protection from IBD. We investigated the ability of NLRX1 to modulate gut pathology, inflammation and immunity by using DSS and CD4+CD45RBhigh transfer colitis models. Colons, spleens, and mesenteric lymph nodes were excised for characterizing immune cell subsets, histological analyses, cytokine and autophagy expression, NF-κB activity, and ROS production. The loss of NLRX1 increased severity of disease and colonic histopathology in both models of IBD. Colons of NLRX1-/- mice had significantly increased epithelial ulceration and leukocyte infiltration in the DSS model, while recipients of NLRX1-/- CD4+ T cells had increased leukocytic infiltration, proliferation, fibrosis, and crypt abscessation in both colon and ileum. The loss of NLRX1 increased numbers of Th1, Th17, and Treg in the spleen, increased colonic NF-κB activity, upregulation of IL-17, IFNγ and TNF-α production, and increased ROS production. NLRX1 ameliorates intestinal pathology during IBD by acting as an internal thermostat that modulates the balance of effector versus regulatory CD4+ T cell responses, and suppressing colonic NF-κB activity, inflammatory cytokine expression, ROS production, and autophagy.
VDJServer: a web-accessible analysis portal for immune repertoire sequence data (HUM1P.317)
Authors: Cowell Lindsay , Fonner John , Jordan Christopher , Levin Mikhail , Mock Stephen , Monson Nancy , Rounds William , Salinas Edward , Scarborough Walter , Scheuermann Richard 
Published: January 2023

Abstract
Despite the importance and widespread application of detailed antigen receptor repertoire profiling via high-throughput sequencing, there is currently no suite of software tools for seamless, reproducible analysis of repertoire sequence data. Tools exist for only a subset of analysis tasks. They do not function together and are difficult to reuse without modification that requires bioinformatics expertise. This leaves researchers to perform repetitive and error-prone tasks by hand, to develop internal, idiosyncratic algorithms not easily generalizable or systematically applied, and to expend significant manual labor reformatting primary and derived data for passing between tools. This approach is not only error-prone and time- and labor-intensive, but it comes at the expense of reproducibility, both within and between research groups. We have addressed this problem by developing VDJServer, a suite of repertoire analysis tools accessible via a web interface that allows users to upload a set of antigen receptor repertoire sequences and pass them through a seamless, customizable workflow that executes all steps in the analysis and generates analysis reports complete with data summary tables, statistical analyses, figures, and workflow logs. VDJServer runs on the high-performance computing resources at the Texas Advanced Computing Center, thus enabling users to process extremely large data sets. VDJServer is funded by the NIAID and is freely available.
Mouse models for immunology research available from the Jackson Laboratory Repository (HUM1P.318)
Authors: Valenzuela Alicia , Fancher Karen , Rockwood Stephen , Sasner Michael 
Published: January 2023

Abstract
The Jackson Laboratory Repository serves as a centralized facility for the development, distribution and cryopreservation of mouse models of human biology and disease. Hundreds of new strains are added annually to one of the largest collections of characterized mouse strains available. This poster presents our newest knockout (KO) strains involving genes associated with cell-mediated and humoral immunity. Lag3 (lymphocyte activation 3) KO mice allow further characterization of Treg suppressive function. Pvr (CD155)-null mice carry a lacZ reporter and exhibit inefficient IgA/IgE responses in response to challenge. The loxP-flanked Bcl6 mice carry a conditional allele involved in the differentiation of CD4 T cells into follicular Th cells. Two strains provide reporter/KO mutants of genes involved in the complement pathway, C1qa and Cfb. In addition, the hematopoietic cytokine, Il3, joins our extensive collection of interleukin mutants. New research tools include the following: a strain with EGFP and cre expression from the Zbtb16 (PLZF) promoter, in which expression is directed to the NKT cell lineage; a cre knockin (KI) into the Cx3cr1 promoter region for fate-mapping studies of the monocyte/macrophage compartment; and a ROSA26 KI that features a two-color fluorescent Cre reporter (widespread red fluorescence prior to cre exposure and localized green fluorescence following cre exposure). Researchers are encouraged to deposit mice to the Repository: www.jax.org/donate-a-mouse.