bluuebunny/crossref_metadata_2025_split huggingface dataset:
Selenoprotein K is required for palmitoylation of CD36 in macrophages: implications in foam cell formation and atherogenesis (P1102)
Authors: Fredericks Gregory , Meiler Svenja , Baumer Yvonne , Huang Zhi , Hoffmann Fukun , Rose Aaron , Norton Robert , Boisvert William , Hoffmann Peter 
Published: January 2023

Abstract
Selenoprotein K (Selk) is an endoplasmic reticulum transmembrane protein important for calcium flux and macrophage activation, but its role in foam cell formation and atherosclerosis has not been evaluated. Bone marrow-derived macrophages (BMDM) from Selk-/- mice exhibited decreased uptake of modified low density lipoprotein and foam cell formation compared to wild-type controls, and the differences were eliminated with anti-CD36 blocking antibody. CD36 expression was decreased in TNFα-stimulated Selk-/- BMDM compared to wild-type controls. Fluorescence microscopy revealed TNFα-induced clustering of CD36 in wild-type BMDM indicative of lipid raft localization, which was absent in Selk-/- BMDM. Immunoprecipitation showed Selk-/- BMDM have decreased CD36 palmitoylation, which is crucial for both stabilizing CD36 expression and directing its localization to lipid rafts. To assess if this phenomenon had a role in atherogenesis, high-fat diet was fed to irradiated Ldlr-/- mice reconstituted with bone marrow from Selk-/- or wild-type mice. Selk was detected in aortic plaques of controls, particularly in macrophages. Selk deficiency in immune cells led to reduction in atherosclerotic lesion formation without affecting leukocyte migration into the arterial wall. These findings suggest that Selk is important for stable, localized expression of CD36 in macrophages during inflammation, thereby contributing to foam cell formation and atherogenesis.
Resveratrol ameliorates high fat diet-induced fatty liver with alteration of Kupffer cell subset in mice. (P1106)
Authors: Nishikawa Kahoko , Kinoshita Manabu , Iwaya Keiichi , Fujiwara Yoko , Sakamoto Toshihisa , Seki Shuji 
Published: January 2023

Abstract
Background: We gave considerable attention to improve a high-fat diet (HFD)-induced fatty liver by Resveratrol from a standpoint of lifestyle disease prevention. Subject: To determine which protein is strongly regulated by Resveratrol in HFD-induced fatty liver, we investigated the modulation of liver protein and Kupffer cell subset in the HFD mice. Material and Methods: Mice (♂C57BL/6, 7 weeks) were fed HFD, HFD supplemented 0.2% Resveratrol or control diet for 8 weeks. Protein extraction from the liver was analyzed by LCMS-based proteomics. Immunostaining was performed using F4/80, CD68, CD11b and adipophilin (ADFP) Abs, and the Kupffer cell subset was analyzed by flow cytometry. TNF production by Kupffer cells was also measured. Results: Resveratrol decreased liver weights and hepatic triglyceride (TG) levels in the HFD mice without changing food intake, and improved fatty liver. Lipid metabolism was strongly down-regulated by Resveratrol. The most down-regulated protein was the ADFP that plays a role of TG storage in lipid droplets and macrophage. Resveratrol reduced lipid droplets and ADFP in the liver of HFD mice. Concurrently, Resveratrol suppressed increased CD68-CD11b+ Kupffer cell subset with suppressing their TNF production in the HFD mice. Conclusion: Resveratrol altered lipid metabolism, especially ADFP, in HFD mice, and suppressed TNF-producing CD11b+ Kupffer cells, thereby ameliorating fatty liver.
Investigating how bacteriophages influence the human immune response to Propionibacterium acnes (P3148)
Authors: Marinelli Laura , Modlin Robert 
Published: January 2023

Abstract
Numerous lines of evidence implicate the Gram-positive skin commensal, Propionibacterium acnes, in the pathogenesis of acne vulgaris, in part by eliciting a host inflammatory response. The emergence of antibiotic resistance in up to 60% of P. acnes isolates, along with the adverse side effects of current treatments has highlighted the need for the development of safer and more effective anti-acne therapeutics. We have investigated the bacteriophages that infect and kill P. acnes and found that they have limited genetic diversity and broad host ranges against clinical isolates, pointing to the potential utility of a phage-based acne treatment. Here, we explored whether these phages, by lysing P. acnes, influence the human immune response to the bacteria. Human monocytes were cultured in the presence of phage-killed or live bacteria, and cytokine responses were measured. We found that monocytes cultured with phage-killed bacteria secrete lower levels of pro-inflammatory cytokines, such as TNFα, IL-1β and IL-12p40, as compared to those infected with live bacteria; phage alone did not stimulate a response. This pattern was also observed when phage and bacteria were added concurrently, that is, during active phage infection, and in all cases, this was consistent across multiple donors. These data suggest that bacteriophage-mediated killing of P. acnes bacteria could help to ameliorate the inflammation associated with acne as a therapeutic modality.
Modulation of mucosal immune responses to Clostridium difficile by probiotic bacteria (P3073)
Authors: Viladomiu Pujol Monica , Hontecillas Raquel , Pedragosa Mireia , Philipson Cassandra , Bassaganya-Riera Josep 
Published: January 2023

Abstract
Commensal bacteria play a critical role in maintaining intestinal epithelium and mucosal immune homeostasis. Alterations in intestinal microflora predispose mammalian hosts to infection with pathogens such as Clostridium difficile, which has recently re-emerged as a pathogen causing nosocomial diarrhea, colitis and death. We investigated the mechanisms by which the probiotic VSL#3 ameliorates C. difficile-associated disease (CDAD) in mice. Oral VSL#3 administration reduced disease activity, body weight loss, and colonic inflammatory lesions after infection. Probiotic treatment also decreased Th17 and IL-17-producing TCRγδ cell populations at the colonic lamina propria as well as production of IL-17 and IL-1β in the colonic mucosa. Increased numbers of regulatory T cells (Treg) were seen in spleens of VSL#3-treated mice, which correlated to increased peroxisome proliferator-activated receptor γ (PPARγ) activity. PPARγ is a transcription factor and nuclear receptor that modulates immune responses and whose activation is essential for the maintenance of innate antimicrobial immunity in the colon. Mice treated with VSL#3 had an upregulation of calprotectin antimicrobial peptides S100A8 and S100A9 in the colon correlating to decreased fecal clostrial burden at day 4 post-infection. VSL#3 probiotic bacteria regulate colitis by increasing PPARγ activity and decreasing IL-17 responses and increase clostridial clearance by upregulating antimicrobial peptides in mice with CDAD.
CD73-generated extracellular adenosine regulates Toxoplasma gondii infection via NKT cell cytokine production. (P3057)
Authors: Toussaint Leon , Mahamed Deeqa , Bynoe Margaret 
Published: January 2023

Abstract
Toxoplasma gondii is an apicomplexa, obligate intracellular pathogen that evades the host immune system-mediated clearance by undergoing stage differentiation to persist indefinitely in susceptible hosts. We have previously shown that mice deficient in the ecto-enzyme CD73, which generates extracellular adenosine, were significantly resistant to acute and chronic toxoplasmosis after oral infection with the T. gondii type II ME49 strain. Resistance in CD73KO mice was due to a defect in parasite differentiation in the central nervous system (CNS). In contrast to oral infection, intraperitoneal infected CD73KO mice were highly susceptible to immune-mediated pathology, with significantly increased infiltration of neutrophils and T cells into the peritoneal cavity. The adenosine receptor agonist NECA offered protection against immunopathology in infected CD73-/- mice, suggesting the absence of CD73-generated adenosine led to increased susceptibility in these mice. Peritoneal exudate cells from infected CD73KO mice generated higher levels of the inflammatory mediators nitric oxide, TNFα, IL1β and IFNγ. Interestingly, despite the high IFNγ in CD73-/- mice, they produced little to no IL-12. We propose that the IFNγ production independent of IL-12 in these mice during T. gondii infection may be produced by NKT cells (proficient producers of IFNγ). Thus extracellular adenosine is a key molecule that regulates the immune response to an intracellular pathogen and promotes host survival.
Vav1 cooperate with Themis to modulate regulatory T cell suppressive functions (P1217)
Authors: Pedros Christophe , Chabod Marianne , Dejean Anne S , Bernard Isabelle , Lagrange Dominique , Andreoletti Olivier , Fournié Gilbert J , Saoudi Abdelhadi 
Published: January 2023

Abstract
Natural Foxp3+ regulatory T cells (Treg) are of paramount importance for establishing self-tolerance, controlling inflammatory responses and maintaining immune homeostasis. The identification of new molecules controlling Treg functions is nowadays an important challenge. We previously reported the implication of Themis, a new actor of TCR signaling, in Treg suppressive capacities. Indeed, Treg from Themis deficient BN rats were defective, leading to the development of an inflammatory bowel disease. In the present study, we show that Themis deficiency does not impair Treg suppressive functions and does not induce any pathological manifestations in LEW rats. These findings suggest that an interaction with other(s) gene(s) may contribute to the Treg defect and disease development in Themis deficient BN rats. To test this hypothesis and identify the involved gene(s), we performed a genetic dissection using different congenic lines. We show that the impact of Themis deficiency on Treg suppressive functions requires the expression of a constitutively active Vav1 variant. Together, these studies highlight the importance of Vav1 and Themis signaling hub in regulating the suppressive functions of Treg. Understanding the underlying molecular mechanisms may lead to advances in Treg biology and favor the identification of new pharmacogenetic markers and therapeutic targets in immune mediated diseases.
T cell-derived inducible nitric oxide synthase switches off Th17 cell differentiation (P1225)
Authors: Zhang Ruihua , Yang Jianjun , Xiong Huabao 
Published: January 2023

Abstract
Although it is clear that RORgt is necessary for the generation of Th17 cells, the molecular mechanisms for the regulation of Th17 cell. here, we show that activation of CD4+ T cells results in the expression of inducible nitric oxide synthase (iNOS), but not endothelial NOS (eNOS) or neuronal NOS (nNOS). iNOS deficient mice displayed enhanced Th17 cell differentiation but without major effects on either Th1 or Th2 cell lineages, while eNOS or nNOS mutant mice showed comparable Th17 cell differentiation compared to wild type control mice. Addition of dihydrochloride (L-NIL), the iNOS inhibitor, significantly enhanced Th17 cell differentiation and S-nitroso-N-acetylpenicillamine (SNAP), the NO donor, dose-dependently reduced the percentage of IL-17-producing CD4+ T cells. NO mediates nitration of tyrosine residues in RORgt leading to the suppression of RORgt-induced IL-17 promoter activation, indicating that NO regulates IL-17 expression at the transcriptional level. Finally, studies of an experimental model of colitis showed that iNOS deficiency results in more severe inflammation with an enhanced Th17 phenotype. These results suggest that NO derived from iNOS by activated T cells plays a negative role in the regulation of Th17 cell differentiation and highlight the important of intrinsic programs for the control of Th17 immune responses.
Impact of age and body fat on increased toxicity following systemic immunotherapy (P1235)
Authors: Mirsoian Annie , Bouchlaka Myriam , Sckisel Gail , Monjazeb Arta , Murphy William 
Published: January 2023

Abstract
Both aging and obesity are associated with increased distribution of visceral fat and the presence of a chronic pro-inflammatory state. Recent studies have shown that strong systemic immune stimulation with combined aCD40 and IL-2 immunotherapy (IT) of aged (&gt;15 mos) mice results in rapid lethality within two days of treatment as opposed to young mice whom are able to tolerate the entire regimen. Aged mice exhibited lethal toxicities via increased pathological levels of serum proinflammatory cytokines (TNFa, IL-6), and increased multi-organ damage to the liver, lung and gut. Liver damage was further evidenced by heightened serum ALT. Given that aging is characterized by an increase in lipid deposition, we assessed the impact of fat content upon IT induced cytokine storm and tissue damage by comparing young obese (ob/ob) mice with aged caloric-restricted (CR) and age-matched control (WT) mice. Young obese mice showed greater serum cytokine levels of TNFa and IL-6 after immune stimulation, similar to aged mice, in comparison to age-matched controls but did not incur multi-organ pathology as seen in the aged cohort. These data demonstrate the intricate relationship between age and body fat suggesting that fat may be responsible for the exacerbated cytokine storm associated with aging but that aging itself may play a unique and distinct role in cytokine induced acute multi-organ toxicities, possibly through differential cytokine receptor expression in comparison to young.
Soluble IP4 limits NK cell effector functions by controlling PI3K signaling (P1198)
Authors: Sauer Karsten , Park Eugene , Siegemund Sabine , French Anthony , Wahle Joseph , Sternberg Luise , Rigaud Stephanie , Jonsson A. , Yokoyama Wayne , Huang Yina 
Published: January 2023

Abstract
NK cells have important functions in cancer immunosurveillance, bone marrow allograft rejection, fighting infections and reproduction. NK cell-based therapies are promising blood cancer treatments. Overcoming their currently limited efficacy requires a better understanding of the molecular mechanisms controlling NK cell development and dampening their effector functions. NK cells recognize pathogen-infected or tumor cells through invariant NK cell receptors (NKR), and then kill such stressed cells. Two second-messenger pathways downstream of NKRs are required for NK cell maturation and effector responses: PIP3-generation by PI3K, and generation of DAG and IP3 by PLCγ. IP3 plays a key signaling role by mobilizing Calcium, but can also be converted into soluble inositol(1,3,4,5)tetrakisphosphate (IP4). We and others previously showed that IP4 can act as a soluble analog of the PI3K lipid-product phosphatidylinositol(3,4,5) trisphosphate (PIP3) and control PIP3-mediated signaling protein membrane recruitment and activation in thymocytes and granulocytes. Here, we show data which suggest a novel IP4 function in promoting NK cell terminal differentiation and acquisition of a mature NKR repertoire. However, in mature NK cells, IP4 limits NKR induced IFNγ secretion, granule exocytosis and target-cell clearance, in part by inhibiting the PIP3 effector-kinase Akt. This identifies IP4 as an important novel regulator of NK cell development and function, and expands our understanding of the therapeutically important mechanisms dampening NK cell responses. Our results further suggest that PI3K regulation by soluble IP4 is a broadly important signaling paradigm.
STAT4 is dispensable for Th17 generation but essential for accumulation of pathogenic CD4 T cells in the CNS during EAE. (P4114)
Authors: McWilliams Ian , Harrington Laurie 
Published: January 2023

Abstract
Multiple Sclerosis is a demyelinating autoimmune disease of the central nervous system (CNS) and is studied using the mouse model experimental autoimmune encephalomyelitis (EAE). Studies utilizing EAE have shown critical pathogenic roles for Th1 and Th17 CD4 T cell subsets. STAT4 is an important Th1 transcription factor that, when activated by IL-12, results in the production of IFNγ, the hallmark Th1 cytokine. Deletion of STAT4 protects mice from EAE; in contrast, deletion of IL-12 and/or IFNγ does not ameliorate disease, revealing gaps in our current understanding of STAT4 biology during EAE. Given that STAT4-/- mice are resistant to EAE, we utilize bone marrow (BM) chimeric mice to provide a physiological environment in which to study the intrinsic role of STAT4 in CD4 T cells during EAE. Interestingly, we find that during EAE the in vivo frequency of STAT4-/- CD4 T cells is greatly reduced in the CNS of BM chimeric mice compared to the wild-type (WT) CD4 T cells. In addition, these STAT4-/- CD4 T cells are able to produce the IL-17 (the hallmark Th17 cytokine) and activate STAT3 via IL-23 signaling (a critical Th17 pathway associated with disease). Although the “pathogenic” Th17 phenotype is intact in STAT4-/- CD4 T cells, this is not sufficient to drive EAE in STAT4-/- mice, therefore, we posit that the defect in CD4 T cell accumulation resulting from either proliferation, cell death, or migration is the critical STAT4 mechanism driving EAE pathogenesis.
In vivo multi-photon molecular imaging technique visualizes immune and inflammatory cell cross-talks in metabolic diseases (P3231)
Authors: Nishimura Satoshi , Nagasaki Mika 
Published: January 2023

Abstract
To elucidate the underlying mechanisms of adult common diseases based on chronic inflammation, it is vital to examine the immune and inflammatory cell kinetics in living animals. Therefore, we developed in vivo imaging technique based on single- and multi-photon microscopy, and we assessed dynamic cellular interplay in diseased conditions. We found that large numbers of CD8+ effector T cells infiltrated into obese adipose, and these cells were essential for the initiation and development of adipose inflammation. In addition, we identified a novel B cell subset that is abundant in adipose tissue. Immunological or genetic B cell depletion, and B cell-specific Il10 deletion enhanced CD8+ T cell activation and adipose inflammation, and exacerbated insulin resistance in obesity. Adipose environmental factors supported B cell survival and IL-10 production. Adipose B cells also exhibited unique surface phenotypes, distinct from those of known regulatory B cell subsets. Our findings indicate that adipose B cells are a naturally occurring regulatory B cell subset that is essential for negative regulation of diet-induced adipose inflammation and maintenance of homeostasis within adipose tissue, and that B cell dysfunction pivotally contributes to the progression of inflammatory processes. Our results also clearly demonstrated the power of our imaging technique to analyze complex cellular interplays in inflammatory diseases, especially parenchymal and stromal cell cross talks, and to evaluate new therapeutic interventions against them.
Global miRNA expression profiles uncover serum miRNAs as novel biomarkers for diabetes staging in NOD mice (P3267)
Authors: Mi Qing-Sheng , Weiland Metthew , Qi Ruiqun , Zhou Li 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is an autoimmune diseases resulting from T cell-mediated pancreatic beta cell destruction. New biomarkers are urgently needed for earlier T1D risk prediction and progression. MicroRNAs (miRNAs) regulate beta cell and immune cell development and function, and are involved in T1D development. Recently, it has been discovered that serum contains large amounts of stable miRNAs derived from immune cells and other tissues/cells, and that serum miRNAs might serve as biomarkers for disease prediction and progression. Yet little is known about serum miRNAs in T1D. We hypothesize that serum miRNAs could be a novel class of blood-based biomarker for diabetes staging. Serum RNAs were isolated from nonobese diabetic (NOD) mice at different stages (3-4, 7-8, 16-19 weeks, and newly diagnosed diabetes) during diabetes development. Serum miRNA expression profiles were performed by real-time RT-PCR MicroRNA Arrays. Twenty-three miRNAs appeared to have specific expression patterns during diabetes development. Single qRT-PCR further confirmed that serum miR-150, miR-146b, and miR-215 are significantly upregulated starting at the stage of insulitis, destructive insulitis and diabetes, respectively. Our data highly suggest that serum miRNAs are potential novel biomarkers for diabetes prediction and staging, and that miR-150, miR-146b and miR-215 could be used as serum biomarkers for diabetes staging or therapy response in the NOD mouse.
Dual analysis of autoantibody and cytokine profiles improves the predictive power of biomarker discovery in lupus (P3277)
Authors: Bourdon David , Van Le Suzy , Liu Xiangdong , Thio Guene , Meng Lihao , Ramachandran Niro 
Published: January 2023

Abstract
We’ve developed ProtoPlex™, a Luminex® xMAP® bead-based assay capable of measuring human autoantibodies to our nearly 9,500 already purified proteins. Combining autoantibody and cytokine profiles yields statistically powerful data sets for disease relevant biomarker discovery efforts. Biomarker discovery workflows fall into two important areas: discovery and validation; yet, no single well-established discovery-to-validation workflow exists. There's growing demand among translational investigators for predictive markers of diseases such as cancer and autoimmune disorders. Clinical research aims to identify proteomic biomarkers leading to early detection and patient stratification, as well as provide prognostic information for guidance and monitoring of treatment efficacy. Given the importance of cytokine signaling in regulation of the immune system, and that aberrant signaling may lead to development of autoimmune diseases, we hypothesized that there is synergistic value in measuring both halves of the proteome in human serum samples: antigens and autoantibodies. Here we describe this novel approach, utilizing the ProtoPlex™Autoantigen Panel and Novex® inflammatory cytokine panels, to identify distinct marker sets for the chronic autoimmune disease Systemic Lupus Erythematosus.
A sensitive immuno-PCR assay to measure a pro-inflammatory cytokine in human patient serum (P3243)
Authors: Khalfin Yana , Asundi Aarati , Delarosa Donnie , DeForge Laura , Nelson Chris , Vandlan Richard , Meng Gloria , Loyet Kelly 
Published: January 2023

Abstract
Pro-inflammatory cytokines may be useful biomarkers for certain therapies in relevant patient samples. However, detection of low abundance cytokines in patient serum has been difficult for some cytokines, thus there is a need to develop a more sensitive assay than the traditional ELISA. Herein we developed an immuno-PCR assay that is in the low to sub pg/ml range to detect levels of a pro-inflammatory cytokine in human serum samples. Immuno-PCR combines the amplification power of PCR with the versatility of ELISA, resulting in improved sensitivity. Biotinylated anti-analyte bound to Streptavidin-coated tubes was used to capture analyte. The captured analyte was detected by anti-analyte conjugated to DNA, in combination with real-time PCR. This immuno-PCR method was found to be at least ~17-fold more sensitive than ELISA, with a lower limit of quantification of ~2 pg/ml in human serum. While this is a promising improvement, a greater fold-improvement over the ELISA detection limit may still be possible. Therefore, we are currently investigating conditions and variables to further improve the sensitivity of this immuno-PCR assay. This assay strategy may be helpful, not only for biomarker quantification, but also in numerous applications where a very sensitive assay is needed.