bluuebunny/crossref_metadata_2025_split huggingface dataset:
The orphan nuclear receptor estrogen-related receptor α negatively regulates Toll-like receptor-induced inflammation through A20 induction (IRM11P.759)
Authors: Kim Tae Sung , Yuk Jae-Min , Jo Eun-Kyeong 
Published: January 2023

Abstract
Estrogen-related receptor (ERR) α, an orphan nuclear receptor member, is a key metabolic regulator in various organs and tissues. However, its function in regulating innate immune responses has remained largely unknown. Herein we show that ERRα negatively regulates toll-like receptor (TLR)-induced inflammatory responses through induction of A20. ERRα-deficient (ERRα-/-) mice showed a higher susceptibility to endotoxin-induced septic shock, with a concurrent increase of proinflammatory responses in vivo and in vitro. Global gene expression analysis showed that ERRα regulated TLR-induced inflammation through induction of A20, a deubiquitinating enzyme in TLR signaling. Lack of ERRα in macrophages increased NF-κB signaling and IKK activation. In addition, treatment of macrophages with inverse agonist XCT-790 led to an increase of inflammatory responses induced by TLR4 stimulation. These findings identify a novel function of ERRα as an intrinsic negative regulator of inflammation through induction of A20.
The role of CD99 in T cell activation, signaling, and immunological synapse formation (IRM11P.760)
Authors: Nam Giri , Choi Eun Young 
Published: January 2023

Abstract
Cytotoxic T lymphocytes (CTLs) provides a defense against virus-infected and tumorigenic cells. The recognition of antigen-presenting cells (APCs) for T cells and the association with them are essential in the formation of the immunological synapse (IS) which triggers T cell activation. Although various proteins that stabilize the IS structure for delivering appropriate signals in T cells have been identified, the role of adhesion molecule CD99 in CTL activation and IS formation has not been investigated. Here we found that CD99 in CTLs participates in T cell activation, proliferation, and effector differentiation by inducing of cluster formation between T cells and APCs. Interestingly, it is shown that CD99 is co-localized more at the peripheral supramolecular activating cluster (pSMAC) region than the central supramolecular activating cluster (cSMAC) in the IS, indicating the involvement of CD99 in IS formation. The absence of CD99 in CD8+ T cells resulted in slower formation of T-APC conjugates than WT CTLs. In addition, Akt-mediated signaling, followed by activation, proliferation, and cytokine production, was delayed in CD99KO CTLs upon TCR stimulation. Results presented here demonstrate for the first time that CD99 is recruited to the IS during APC and T cell interaction and plays an important role in subsequent T cell activation.
CD28 induces mitochondrial respiration in long lived plasma cells for reactive oxygen species dependent survival and NFκB-mediated Blimp1 expression. (LYM6P.771)
Authors: Utley Adam , Rozanski Cheryl , Nair Jayakumar , Lee Kelvin 
Published: January 2023

Abstract
Sustained humoral immunity is dependent upon bone marrow resident long-lived plasma cells (LLPCs). We have shown that CD28 signaling in LLPCs is necessary for their survival and antibody production; however, the signaling pathways that define LLPC survival and function are not completely understood. In T cells, CD28 is known to induce glycolysis at the expense of mitochondrial respiration. To our great surprise, CD28 induces mitochondrial respiration-dependent reactive oxygen species (ROS) through PLCγ1 in LLPCs but not splenic short lived plasma cells (SLPCs). Paradoxically, inhibition of ROS kills LLPCs while promoting survival of SLPCs. In LLPCs, CD28-mitochondrial ROS activates NFκB, which in turn binds directly to the Blimp1 promoter, the master regulator of plasma cell identity. Furthermore, CD28 induces Blimp1 expression in LLPCs but not SLPCs. These data suggest a model wherein CD28-induced mitochondrial respiration drives survival of LLPCs through mitochondrial ROS-dependent activation of NFκB and subsequent Blimp1 upregulation. By understanding CD28 regulation of LLPC metabolism, new therapeutic targets arise for antibody-mediated autoimmune disease and vaccine design.
Differential regulation of pro-inflammatory cytokine expression by MAP kinases in macrophages in response to intestinal parasite infection (MPF3P.816)
Authors: Lim Mei Xing , Jiao Huipeng , Png Chin Wen , Tan Shyong Wei Kevin , Zhang Yongliang 
Published: January 2023

Abstract
Blastocystis is a common enteric parasite that is associated with inflammatory bowel disease. However, the pathogenic status of Blastocystis remains unclear. In this study, we found that Blastocystis induced expression of pro-inflammatory cytokines including IL-1β, IL-6 and TNF-α in murine colon and macrophages. We showed that Blastocystis also induced the activation of the three major groups of MAP kinases (ERK, JNK, and p38) in macrophages. The use of MAPK-specific inhibitors in this study demonstrated differential regulation of pro-inflammatory cytokines in macrophages by MAPKs in response to Blastocystis. We found that ERK activation is important for the expression IL-6 and TNF-α at both mRNA and protein levels and the mRNA expression of IL-1β in response to Blastocystis stimulation. Inhibition of JNK significantly suppressed all three pro-inflammatory cytokines at both mRNA and protein levels. p38 inhibition only suppressed IL-6 secretion but not IL-1β and TNF-α. Furthermore, we found that serine proteases from Blastocystis are important for the activation of MAPKs and cytokine expression in macrophages. Our study demonstrated for the first time that Blastocystis could induce the expression of various pro-inflammatory cytokines via activation of MAP kinases, and infection with Blastocystis may contribute to the pathogenesis of inflammatory intestinal diseases through the activation of inflammatory pathways in host immune cells such as macrophages.
Nanogel-based PspA nasal vaccine induces microRNA-associated protective immunity in nonhuman primates (MUC4P.834)
Authors: Fukuyama Yoshiko , Yuki Yoshikazu , Katakai Yuko , Takahashi Haruko , Sawada Shinichi , Joo Sunyi , Park Eun Jeong , Akiyoshi Kazunari , Kiyono Hiroshi 
Published: January 2023

Abstract
We previously demonstrated a cationic type of cholesteryl group-bearing pullulan based nanogel containing pneumococcal surface protein A (PspA-nanogel) induced both protective Th2-mediated Ag-specific systemic and mucosal antibody (Ab) responses and Th17 cell-mediated immunity. In this study, we examined whether PspA-nanogel nasal vaccine could induce PspA-specific protective immunity and cytokines-related miRNA expression in nonhuman primates. When cynomolgus macaques were nasally immunized with 25 μg of PspA-nanogel/dose, increased levels of Th2 cell dependent PspA-specific serum and broncho alveolar lavage fluid (BALF) IgG, and nasal wash (NW) secretory IgA (SIgA) Ab responses with elevated Th17 cell immunity were seen when compared with control macaques nasally immunized with 25 μg of PspA alone or PBS. MicroRNA (miRNA) analysis of serum and nasal tissues from these PspA-nanogel immunized macaques revealed specific elevation of miR-181a and miR-326 which have been shown to corresponding to Th2 and Th17 responses, respectively. These results suggest that these two miRNAs are also associated with non-human primate Th2 and Th17 cell responses after nasal vaccination with PspA-nanogel vaccine which accounted for the induction of protective immunity against pneumonia.
Mechanism by which commensal bacteria limit inflammation (MUC4P.844)
Authors: Paynich Mallory , Jones Sara , Knight Katherine 
Published: January 2023

Abstract
Trillions of bacteria live in homeostasis within the gastrointestinal tract and provide a variety of benefits to the host immune system. One beneficial function of the healthy microbiota is to protect the host from enteric pathogens; however, little is known about the mechanisms by which this occurs. We utilize a mouse model in which a single dose of the commensal bacterium, Bacillus subtilis, protects mice from disease induced by the enteric pathogen Citrobacter rodentium. Our goal is to elucidate the mechanism by which B. subtilis protects from inflammation caused by infection with C. rodentium. We identified exopolysaccharides (EPS) to be the active molecule of B. subtilis, and a single i.p. dose of EPS protects mice from infection. Because protection occurs within a few days, we hypothesized that EPS elicits a protective response by modulating the innate immune system. EPS binds F4/80+CD11b+ peritoneal macrophages, and after i.p. injection of EPS, we identified F4/80+CD11b+ cells that are MHCII+CD206+, indicative of alternatively activated macrophages. We further show that peritoneal macrophages from EPS-treated mice suppress T cell proliferation in vitro, suggesting that these cells contribute to EPS mediated protection from inflammation. We are testing if these cells differentiate in the peritoneum following EPS treatment and whether they mediate protection from C. rodentium induced inflammation.
Immune modulatory mechanisms of Oncostemum bojerianum extracts during Clostridium difficile infection in mice (MUC4P.845)
Authors: Viladomiu Monica , Hontecillas-Magarzo Raquel , Kingston David , Eaton Alex , Zhang Xiaoying , Philipson Casandra , Schiff Eric , Eden Kristin , Lu Pinyi , Carbo Adria , Bassaganya-Riera Josep 
Published: January 2023

Abstract
Activation of Peroxisome Proliferator-Activated Receptor γ has proven beneficial for the treatment of several inflammatory diseases. However, current drugs targeting PPAR result in significant side effects. Therefore, the development of safer anti-inflammatory therapies is important. This project aimed to discover new natural constituents from ethnomedicinal plants that target PPARγ and elucidate the mechanisms of action underlying their anti-inflammatory activities. From a screen of 200 plant extracts, we identified more than 50 that activate PPARγ, with a high predominance in Oncostenum species. The anti-inflammatory properties of two O. bojerianum extracts were further tested in a mouse model of Clostridium difficile infection, which resulted in a detectable improvement in disease activity, weight loss, colonic inflammatory lesions, and increased levels of colonic IL-10-producing CD4+ T cells. Oral treatment with Oncostemum extracts increased PPARγ and PPARα activity and elevated transcriptional expression of their responsive genes. Bioassay-guided fractionation studies in combination with in silico molecular screening, identified the resorcinol family as the PPARγ-activating compounds contained in the extracts. Characterizing the modulation of cellular and molecular signatures by resorcinols at the sites of inflammation will translate natural products discovery into the development of safer and more efficacious therapeutics for infectious and immune-mediated diseases.
Segmented filamentous bacteria induce systemic IgA responses to commensal bacteria (MUC4P.854)
Authors: Wilmore Joel , Sonnenberg Gregory , Artis David , Allman David 
Published: January 2023

Abstract
IgA is a component of the mucosal barrier surface and plays an important part in maintaining separation between the host and the commensal microbiota. Mucosal IgA responses are considered short-lived and restricted to mucosal sites with no systemic dissemination. However, there is little known about the regulation and maintenance of IgA secreting plasma cells (PCs) and how they compare to bone marrow PCs. Interestingly, we find that IgA secreting PCs make up a large proportion of the bone marrow PC pool in mice colonized with segmented filamentous bacteria (SFB), but not in germ-free or conventional SFB free mice. The increase in IgA PCs in the bone marrow correlates with a large increase in serum IgA. The co-housing of SFB free mice with SFB colonized mice results in significantly increased levels of IgA in the serum and higher numbers of IgA+ PCs in the bone marrow. This process is dependent upon T cells, because SFB colonized TCRβ-/-δ-/- mice have similar levels of IgA bone marrow PCs as wild-type SFB free mice. The SFB associated increase of IgA antibody titers coincides with the ability to detect commensal bacteria specific IgA in the serum. These data suggest that the induction of systemic IgA is influenced by the composition of the commensal microbiota. These results highlight the need for greater understanding of the role commensal bacteria play in systemic immunity, which could impact oral vaccine development and autoimmunity.
Phosphoinositide-3-kinase p110δ isoform promotes influenza pathogenicity (MUC5P.858)
Authors: Hancock Aidan , Boesteanu Alina , Chroneos Zissis , Turner Martin , Katsikis Peter 
Published: January 2023

Abstract
We have found that mice deficient in a tissue specific isoform of the signaling enzyme phosphoinositide-3 kinase (PI3K) p110δ are protected from PR8 influenza virus infection and have reduced weight loss and lung viral loads post-infection. Lung inflammation and immune cell infiltration is also decreased after viral challenge. In addition, p110δ KO mice had fewer influenza virus infected alveolar epithelial cells when infected with a GFP-flu virus and GFP expression was measured in lung epithelial cells. Cytokine measurements in bronchoalveolar lavages suggest p110δ KO mice exhibit increased IFN-gamma and MIG post PR8 infection. Slightly increased numbers of NK cells are detected in the lungs of p110δ KO mice as early as day 3 post-infection however activation was decreased within this population as compared to controls, suggesting they are not the primary source of resistance to influenza. In order to determine which cell type was causing the decreased pathogenicity we used two different tissue specific knockout mouse lines. Mice with a myeloid specific KO of p110δ showed similar weight loss and viral loads to C57BL/6 controls, suggesting alveolar macrophages are not primarily responsible for the observed protection in p110δ KO mice. However, mice with alveolar epithelial cell (AEC) specific deletion of p110δ were protected from influenza virus challenge. These data suggest PI3K signaling cascade is important for influenza virus protection within AEC.
Respiratory infection alters archetypal anti-viral CD8 T cell differentiation programs resulting in diminished memory potential (MUC5P.861)
Authors: Shane Hillary , Verbist Katherine , Klonowski Kimberly 
Published: January 2023

Abstract
Our understanding of memory CD8 T cells has been largely derived from acute, systemic infection models. However, memory CD8 T cells generated from mucosal infection exhibit unique properties and, in respiratory infections, are poorly maintained long-term. To better understand how infection route modifies memory differentiation, we compared murine CD8 T cell responses to an identical VSV challenge generated intranasally (IN) or intravenously (IV). IN infection resulted in greater peak expansion of VSV-specific CD8 T cells. However, this numerical advantage was rapidly lost during the contraction phase of the immune response, resulting in numerical memory CD8 T cell deficiencies when compared to IV VSV infection. Interestingly, the anti-viral CD8 T cells generated in response to IN VSV exhibited a biased and sustained proportion of early effector cells (CD127loKLRG1lo) akin to the CD8 developmental program favored after IN influenza infection, suggesting that respiratory infection favors an incomplete memory differentiation program. Correspondingly, IN infection resulted in lower CD122 expression and EOMES levels by VSV-specific CD8 T cells, further indicative of an inferior transition to bona-fide memory. Together these data suggest that the environments encountered in distinct immunization routes are sufficient to modulate both the quality and quantity of anti-viral CD8 T cells in response to an identical pathogen and should be considered in CD8-based vaccine design.
Accurate quantitation of proteins involved in autoimmune disease using simple western (TECH2P.872)
Authors: Tu Andrea , Nguyen Uyen , Kazakova Irina , Ramirez Francisco , Xu Hui , Proctor John , Boge Annegret 
Published: January 2023

Abstract
Why do researchers use Western blots? Since the technique was developed thirty years ago it’s become a trusted method for confirming the presence or absence of a protein but lacks the quantitative rigor for accurate measurement of the amount of protein present. The process requires many steps which introduces multiple variables. While portions of the technique have been automated to improve consistency, until now there has been no major leap in the technology that propels this method of protein analysis from qualitative to quantitative. Simple Western is the modern evolution of traditional immunoassay techniques. Wes, the latest addition to the Simple Western platform, is an easy to use, fully automated system that removes the variability seen with traditional Westerns for more reproducible results run to run and between users. Researchers can now identify their protein and achieve reliable quantitation of their target proteins. To demonstrate the precision and data reliability of Wes we look at proteins indicative of autoimmune disease in human samples from either normal individuals or individuals suffering from the disease. Wes not only reduces the hands on time and the time to results, but generates highly reproducible data that gives researchers a high degree of confidence in the differences detected between the two disease states. Accurate quantitation of these proteins can lead to the identification of predictive biomarkers for either diagnosis or response to treatment.
Anti-murine CD52 therapy provides anti-inflammatory and neuroprotective effects in an EAE mouse model (THER3P.888)
Authors: Turner Michael , Chretien Nathalie , Havari Evis , Huang Jianhua , Pang Petti , LaMorte Michael , Roberts Bruce , Kaplan Johanne , Siders William 
Published: January 2023

Abstract
To better understand the mechanism of action of alemtuzumab in multiple sclerosis (MS), we have investigated the effects of anti-mouse CD52 antibody (anti-muCD52) treatment in an EAE mouse model. Therapeutic administration of anti-muCD52 antibody significantly reduced the severity and progression of disease. Treatment benefit was associated with a reduction in the number of lymphocytes in the spleen and CNS as well as a reduction in MOG35-55 auto-reactive T cells that produce IFNγ or IL-17 following ex vivo restimulation. Neuroprotective effects were also observed following antibody treatment. Specifically, anti-muCD52 treatment reduced the level of white matter damage in the CNS as determined by histological analysis of myelin basic protein and by measurement of serum phosphorylated neurofilament heavy chain. In addition, axonal conductance of the spinal cord was assessed through electrophysiological measurement of motor-evoked potentials. Animals treated with anti-muCD52 demonstrated higher peak amplitude as well as reduced peak latency compared to control animals treated with vehicle. Collectively, these results suggest that the therapeutic benefit of anti-muCD52 treatment in EAE can be attributed in part to a reduction in the number of auto-reactive T cells, decreased CNS inflammation, and protection of CNS integrity.
Generation and characterization of a mouse LIGHT specific inhibitor in a murine model of colitis (THER4P.891)
Authors: Felix Nathan , Kansal Selena , Stetsko Dawn , Yang XiaoXia , Xie Jenny , Suchard Suzanne 
Published: January 2023

Abstract
LIGHT (TNFSF14, CD258) is a member of the TNF superfamily that is expressed on activated T cells, immature DCs, macrophages, and NK cells. LIGHT interacts with its receptors (HVEM and LTβR) and is reported to provide co-stimulatory and pro-inflammatory signals to T cells, B cells, DCs, and macrophages. Numerous studies have implicated the pro-inflammatory effects of LIGHT as an important contributor to disease severity in models of inflammatory bowel disease and fibrosis. However, the inhibitors used in many of these studies (e.g. LTβR-Ig and HVEM-Ig) are not specific for LIGHT, and are likely to impact other pathways such as LTαβ and BTLA. Our focus was to develop domain antibodies (dAbs) that specifically block LIGHT-HVEM and LIGHT-LTβR interactions to better understand the therapeutic potential of LIGHT specific blockade. We were able to develop a mouse LIGHT specific inhibitor that potently blockades LIGHT-HVEM and LIGHT-LTβR interactions in vitro. Notably, this dAb had little effect when used as a monotherapy in a murine model of colitis, but when combined with CD40 blockade showed a high degree of efficacy (equivalent to CTLA4-Ig treatment). These data suggest that LIGHT monotherapy may not be sufficient to impact disease severity, but LIGHT blockade may have potential as part of a combination approach.
VNN2 is a new surface marker for monocytic human myeloid-derived suppressor cells (TUM4P.900)
Authors: Soler David , McCormick Thomas , Cooper Kevin , Young Andrew 
Published: January 2023

Abstract
Studies of myeloid-derived suppressor cells (MDSC) have been hampered by the scarcity and lack of cell-specific markers. Specifically, monocytic MDSC (M-MDSC) are currently characterized as CD14+ HLA-DR-/low and only the absence or low expression of HLA-DR is used as a marker to specifically isolate and purify live M-MDSC. However, identifying cells based on the absence or reduce expression of a protein significantly hampers their study. Reviewing previous studies regarding monocytic markers, we noted Vanin-2 as a protein that has been described in the past to be present in monocytic subsets displaying reduced antigen presentation as well as high reactive oxygen production, hallmarks of MDSC. In order to determine if this marker was present on cells we currently classify as M-MDSC, we used flow cytometry to isolate MDSCs and compared by qPCR CD14+ HLA-DR-/low (M-MDSC) versus CD14+ HLA-DR+ monocytes for VNN2 expression. Interestingly VNN2 showed a 12.43 fold increase in CD14+ HLA-DR-/low M-MDSC compared to CD14+ HLA-DR+ monocytes. Next, we sorted CD14+ VNN2+-expressing monocytes (&gt;10%) and we performed a suppressive assay with autologous bead-stimulated CD8 T-cells. This assay showed CD14+ VNN2+ monocytes suppressive capabilities were comparable to those of traditional CD14+ HLA-DR-/low M-MDSC at 24%, compared to CD8 T-cells alone (90%) or CD14+ VNN2neg monocytes (~70%). These results suggest VNN2 might be a viable surface marker to positively identify live human M-MDSC.