bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intestinal mucus-derived nanoparticles carry prostaglandin E2 and suppress liver inflammation through Wnt/β-catenin signaling (P3294)
Authors: Deng Zhong-bin , Zhuang Xiaoying , Xiang Xiaoyu , Jiang Hong , Wang Qilong , Miller Donald , Zhang Huang-ge 
Published: January 2023

Abstract
The Wnt/β-catenin pathway has been known to play a role in induction of immune tolerance, but its role in the induction and maintenance of NKT cell anergy is unknown. We found that activation of the Wnt pathway(s) in the liver microenvironment is important for induction of NKT cell anergy. We identified Glycolipid α-GalCer treatment of mice induced the expression of wnt3a and wnt5a in the liver, and subsequently resulted in a liver microenvironment that induced NKT cell anergy to α-GalCer restimulation. Mice treated with PGE(2) associated with intestinal mucus-derived exosome-like nanoparticles (IDEN) induced NKT cell anergy. PGE2 treatment leads to activation of the Wnt/β-catenin pathway by inactivation of GSK-3β of NKT cells. Pro-inflammatory conditions enhanced the migration of IDEN to the liver where both α-GalCer and PGE2 induced activation of the Wnt pathway thereby inducing NKT anergy in response to subsequent α-GalCer stimulation. IDEN-associated PGE2 also induces NKT cell anergy through modification of the ability of DCs to induce IL-12 and IFN-β in the context of both glycolipid presentation and TLR-mediated pathways. These findings demonstrate that IDEN associated PGE2 serves as an endogenous immune modulator between the liver and intestines and maintains liver NKT cell homeostasis and this finding has implications for development of NKT cell-based immunotherapies.
The anti-oxidative role of glutathione s-transferase mu 2 in anti-GBM induced glomerulonephritis by inhibiting inflammation and oxidative stress (P5121)
Authors: Li Yajuan , Yan Mei , Du Yong , Min Soyoun , Mohan Chandra , Li Quanzhen 
Published: January 2023

Abstract
Oxidative stress is a common manifestation in chronic inflammation and closely associated with autoimmune diseases. Impaired balance between oxidative stress and antioxidant systems exhibits an important impact on pathogenesis of immune-mediated nephritis. Using anti-GBM induced nephritis mouse model, we have identified a group of redox related genes dysregulated in mouse kidney upon anti-GBM antibody challenge. In this study, we investigated the anti-oxidative effect of glutathione s-transferase mu 2 (Gstm2) on immune-mediated nephritis. Human GSTM2 gene was transduced into mesenchymal stem cell (MSC) and hGSTM2-MSCs were injected to anti-GBM antibody challenged 129/svj mice. Mouse blood urea nitrogen (BUN) and proteinuria were measured to evaluate renal function change and renal histopathological changes were appraised. We found the Gstm2 could alleviate the renal inflammatory damage by suppressing expression of inflammatory chemokines, CCL2, IL1β and IL6 to reduce macrophage and T lymphocyte infiltration. Gstm2 improved renal function by reducing BUN and proteinuria. Moreover, hGSTM2-MSCs significantly reduced the apoptosis of tubular cell by regulating the expression of anti-apoptotic genes (BCL2 and CD40LG) and hGSTM2-MSCs resisted oxidative stress by increasing the expression of catalase and glutathione peroxidase 1. In summary, Gstm2 exhibits protective effect on immune-mediated nephritis by inhibiting inflammatory damage and oxidative stress during inflammatory process.
Toll-like receptor 10: its expression and role in chemotaxis in human neutrophils (P1234)
Authors: Singh Baljit , Balachandran Yadu , Knaus Steven 
Published: January 2023

Abstract
TLR10 is a member of TLRs that recognize pathogen-associated molecules and play critical roles in host defense. However, very little is known about the expression and function of TLR10. We found TLR10 expression in vascular endothelium of normal and inflamed lungs from chickens and humans. Western blots showed increased TLR10 protein in lungs of chicken infected with E. coli or Adenovirus. Human neutrophils challenged with E. coli lipopolysaccharide (LPS) showed decreased total TLR10 protein and surface expression in 90 minutes. Confocal microscopy detected cytosolic and nuclear TLR10 in normal neutrophils. Upon activation with LPS, TLR10 colocalized with TLR4, flotallin-1, a lipid raft marker, and EEA-1, an early endosomal marker, to suggest the cycling to endocytic compartments. TLR4 neutralization reduced cytoplasmic localization of TLR10 in neutrophils. Depletion of reactive oxygen species (ROS) with FCCP in LPS-treated neutrophils reduced p65 nuclear translocation and TLR10 expression. Single cell video imaging of live LPS-activated human neutrophils showed the translocation of TLR10 to the leading edge and siRNA-mediated silencing of TLR10 in HL-60 cell line reduced their chemotaxis towards fMLP. We conclude that TLR10 expression is modulated via TLR4 signalling and ROS production, and that TLR10 participates in neutrophil chemotaxis. Altered TLR10 expression in activated neutrophils and inflamed lungs suggests a role for this receptor in lung inflammation.
Differential requirements of CD14 and Lipopolysaccharide Binding Protein in Monophosphoryl lipid A-mediated TLR4 activation (P1209)
Authors: Tanimura Natsuko , Miyake Kensuke 
Published: January 2023

Abstract
Toll-like receptor 4 (TLR4) ligation causes multiple immune responses to host, including inflammations, type-I interferon productions and acquired immune responses. There are a variety of ligands against TLR4, and TLR4 senses those impalpable differences with its co-receptor MD-2. Since TLR4 agonist has highly immunomodulatory functions, it would be useful as an immunostimulator when and if we can harness the responses’ balance. Monophosphoryl lipid A (MPL), one of the artificial derivatives, lacks 1- phosphate and is well-known detoxified agonist. Because of its low inflammatory aspect, MPL has been used for a vaccine adjuvant ingredient. Previous studies reveal that MPL-activated TLR4 transduces its signal by p38-dependent and TIR-domain-containing adapter-inducing interferon-β (TRIF)-biased signaling, though the mechanisms how TLR4 discriminates of MPL has been unclear. Here we focused on the phases of ligand coming into TLR4 and found specific requirement of CD14 and Lipopolysaccharide binding protein (LBP). This unique TLR4 activation induced impaired receptor dimerization and subsequent low inflammatory responses. Our data suggested that the modification of TLR4 activation manners changed responses’ balance.
Simple and robust whole blood staining procedure using PerFix-nc reagents generates unprecedented FOXP3 signal/noise ratio. (P1053)
Authors: Malergue Fabrice , Khemici Laeticia , Montero-Julian Felix A. 
Published: January 2023

Abstract
FOXP3 is a nuclear antigen expressed specifically in regulatory T cells (Tregs). Tregs play critical role in the maintenance of the dominant self-tolerance and have been subject of extensive research effort for the last two decades. Today the detection of FOXP3 is laborious, time consuming (3-4 hours) and poorly reproducible. A new commercially available method, named PerFix-nc, allows simultaneous intra- and extra-cellular staining without any wash step. This new kit allows for the reduction of the workflow to 45 min. In this study we aimed at the optimization of the PerFix-nc protocol for the detection of FOXP3. We evaluated various clones from various vendors, optimizing titration and incubation time, and adding some extra washing steps. Signal to noise ratio at least 40% better than the one obtained for the same samples with reference procedure. Improved cell scattered separation compared to reference method was revealed. Purity and recovery were also significantly increased. Furthermore the method was very robust and demonstrated excellent repeatability with respect to percentage of FOXP3-positive cells and signal to noise. Total workflow was reduced from 3-4 hours in reference method to about 95 min. In conclusion, the novel PerFix-nc reagent significantly improves FOXP3 detection in several different ways: it simplifies FOXP3 intracellular staining procedure, generates robust and highly reproducible results, and provides unprecedented signal to noise ratio.
Ubiquitous MHC class II peptides shape regulatory T cell development (P1039)
Authors: LeGuern Christian , Germana Sharon 
Published: January 2023

Abstract
T cell auto- and allo- reactivity is primarily tamed in the periphery by thymus-derived CD4 regulatory T cells (Tregs) that mature in the medullar compartment. We have achieved Treg-dependent tolerance of fully allogeneic transplants via the transfer of donor MHC class II genes (MHCII) that induced newly made Tregs specific of donor MHCII peptides. Given that gene therapy for tolerance was effective with MHCII but not MHCI genes, the goal of the study was to identify natural MHCII peptides able to impact Treg development and function. Using experimental models that either recapitulated MHCII peptide presentation - the IEα 52-68 peptide presented on I-Ab molecules - or promoted Treg-mediated tolerance to heart grafts via MHCII gene transfer, we confirmed that high amounts of IEα peptide were presented by MHCII from most if not all I-Ab+ CD11c+ dendritic cells of the thymic medulla, suggesting its role in Treg maturation. This hypothesis was confirmed by demonstrating that the introduction of the IEα peptide through hematopoietic chimerism converted host IEα-specific CD4 T cells into CD25hiFoxp3+ suppressive Tregs. Treg suppression for tolerance to transplants required prior activation by cognate pMHCII complexes as MHCII-treated recipients rejected MHCII-deficient while accepting MHCII-sufficient grafts. These data are consistent with a prominent role of “MHCII self-presentation” in shaping thymic Treg TCR specificities and controlling Treg function in the periphery.
STING ligand c-di-GMP improves vaccine efficacy in metastatic breast cancer model (P4265)
Authors: Chandra Dinesh , Asafu-Adjei Denise , Quispe Wilber , Jahangir Arthee , Ramos Ilyssa , Karaolis David , Gravekamp Claudia 
Published: January 2023

Abstract
Cancer vaccination or immunotherapy may be our best and most benign option for preventing or curing metastatic cancer. However, a real breakthrough is hampered by immune suppression in the tumor microenvironment (TME). Myeloid- derived suppressor cells (MDSC) are one of the most important players in immune suppression in the TME. In the study presented here we found that c-di-GMP (ligand for STING), an ubiquitous bacterial signaling molecule and promising vaccine adjuvant, improved the therapeutic efficacy of an attenuated L. monocytogenes-based vaccine expressing tumor-associated antigen (TAA) Mage-b, in a mouse model of metastatic breast cancer (4T1), by targeting MDSC. An extreme low dose of c-di-GMP (0.01 nmol) combined with a low dose of Listeria-Mage-b (104 CFU), dramatically reduced the number of metastases, in correlation with a significant reduction of MDSC and a significant increase in CD8 T cell responses to Mage-b. Moreover, low dose of c-di-GMP increased the production of IL-12, while high dose killed tumor cells directly. Administration of one high dose c-di-GMP to kill the tumor cells, followed by repeated administrations of low dose c-di-GMP to improve CD8 T cell responses, provided similar efficacy against the metastatic breast cancer as Listeria-Mage-b and c-di- GMP in the 4T1 model. These results demonstrate that c-di-GMP is an attractive adjuvant for cancer therapy.
In situ immune modulation therapy with a novel chimeric chaperone generates a potent antitumor response (P4271)
Authors: Yu Xiaofei , Guo Chunqing , Yi Huanfa , Fisher Paul , Subjeck John , Wang Xiang-Yang 
Published: January 2023

Abstract
The immunosuppressive tumor microenvironment (TME) represents a major hurdle for cancer immunotherapy. In the present study, we evaluate the feasibility of conditioning the immune environment in the tumor sites for induction of effective antitumor immunity using a novel immune modulator Flagrp170. This chaperone-based immune modulating agent is created by integrating a pathogen (i.e., flagellin)-derived ‘danger’ signal into Grp170, the largest chaperone resident in the endoplasmic reticulum. This chimeric chaperone molecule is highly efficient in inducing functional activation of dendritic cells via engaging the NF-κB and MAPK signaling pathways, and concurrently facilitating cross-presentation of tumor antigen. Intratumoral delivery of adenoviruses encoding Flagrp170 provokes a strong tumor-specific cytotoxic T lymphocyte response, leading to profound inhibition of tumor growth and metastasis. In situ Flagrp170 therapy preferentially drives the Th1 polarization of the TME, as evidenced by marked elevation of IL-12 and IFN-γ as well as increased tumor infiltration by CD8+ cells. Antibody depletion and blocking assays reveal that CD8+ cells and IFN-γ are essential for enhanced antitumor activity of Flagrp170. Our data demonstrate that this modified chaperone represents a promising immune modulating agent that may be utilized to potentially reverse immunosuppression in the TME and improve immunotherapeutic response to metastatic malignancies.
Use of HSP90 inhibitors in the combination immunotherapy of melanoma (P4272)
Authors: Raveendran Subhara , Rao Aparna , Storkus Walter 
Published: January 2023

Abstract
The molecular chaperone, HSP90 is commonly overexpressed in human melanomas, where it serves to stabilize a broad range of client proteins providing growth, survival and migratory advantages to tumor cells and conditions a pro-angiogenic tumor microenvironment (TME). HSP90-mediated stabilization also limits the proteasome-dependent processing of client proteins and the subsequent presentation of derivative peptides in tumor cell-associated MHC class I complexes which is required for effective host CD8+ T cell recognition of melanoma cells. As a consequence, we have analyzed the anti-tumor impacts of clinical-grade HSP90 inhibitors (i.e. STA9090, 17-DMAG, Novobiocin) on client protein expression in vitro (human melanoma cell line Mel526, murine melanoma cell line B16) and in vivo (s.c. B16 tumors in C57BL/6 mice). We observed that HSP90 inhibitors promote the proteasome-dependent (i.e. blocked by inhibitors PS-341 and MG-132) degradation of client proteins such as c-MET, EphA2, HER2/neu, PDGFRβ and Survivin, with STA9090 appearing to be most efficient on a molar basis. We are currently investigating the capacity of STA9090 to therapeutically sensitize established melanomas to client protein-specific CD8+ T cells (induced by active vaccination or provided via adoptive cell transfer; ACT) in combination therapy approaches. We believe such protocols may hold translational promise for the treatment of patients with advanced-stage melanoma.
IL-9 strategy to perturb Treg function and enhance anti-tumor immunity (P4273)
Authors: Hoelzinger Dominique , Dominguez Ana , Pollock Kevin , Lustgarten Joseph , Cohen Peter , Gendler Sandra 
Published: January 2023

Abstract
IL-9 is a cytokine that is primarily associated with mast cells and airway inflammation, however it also plays a role in T regulatory cell (Treg) survival and recruitment to tumors. We previously reported that 4-1BB treatment of Tregs inhibits both Treg function and IL-9 secretion. Neutralization of IL-9 with an anti-IL-9 antibody inhibits the suppressive function of Tregs without affecting the function of CD4+ and CD8+ T effector cells. Furthermore, the combination of intra-tumoral CpG and anti-IL-9 induced tumor rejection in BALB-neuT and MUC1 tolerant transgenic mice. Here we show that the lack of IL-9 inhibits Treg suppression of T cell proliferation, using IL-9 knockout (IL-9ko) mice. Phenotypic analysis of Treg markers showed that tumor draining lymph nodes of 4T1 bearing mice contain fewer CTLA-4+ and CD103+ Tregs, and that these also have reduced of CTLA-4 and CD103 expression. We also observed that IL-9ko mice spontaneously reject TUBO and 4T1 mammary carcinoma cells and that rejection is abrogated by CD8+ T cell depletion. Finally, preliminary studies of human CD4+ T cell proliferation revealed that IL-9 neutralization led to greater proliferation than isotype control treated cells. Taken together our results suggest that IL-9 can have an immunosuppressive function that comes into play in the tumor microenvironment, and that the blockade of IL-9 could serve as a novel strategy to perturb the function of Tregs to enhance the antitumor effect of tumor vaccines.
Molecular characterization and antimicrobial susceptibility of canine Staphylococcus pseudintermedius isolates from the United States (P6063)
Authors: Videla Ricardo , Bemis David , Kania Stephen 
Published: January 2023

Abstract
Staphylococcus pseudintermedius, the primary cause of canine pyoderma, is therapeutically challenging due to a high, and increasing, rate of resistance to antibiotics. The objectives of this study were: to investigate the genetic diversity within canine S. pseudintermedius isolates in the United States and to characterize their antimicrobial susceptibility profile. Samples from 221 dogs were obtained from veterinary diagnostic facilities located in different regions of the United States. Genetic groups were generated based on Pulsed-field gel electrophoresis (PFGE) and Multi-Locus Sequence Typing (MLST) results. Conventional or real-time PCR was performed to detect presence of the mecA gene as an indicator of methicillin resistance, and the Kirby-Bauer disk diffusion method was used to test the susceptibility to 13 antimicrobials. An association between the largest clonal group and the most commonly encountered susceptibility pattern was found indicating the presence of one major clonal population. These findings represent a significant contribution for the better understanding of S. pseudintermedius genetic diversity in the United States and also reflect the high degree of antimicrobial resistance. Our findings emphasize the need for the development of novel therapeutic alternatives, such as vaccines, to address the increasing number of S. pseudintermedius isolates resistant to the most clinically useful antimicrobials that are available to veterinarians.
Remarkable conservation of natural killer receptor, NKp30 (P6089)
Authors: Ota Yuko , Flajnik Martin 
Published: January 2023

Abstract
Comparative analysis of natural killer receptor (NKR) among vertebrates have revealed that they are remarkably plastic; some receptor families have expanded greatly, while others have been lost in a species-specific manner. Thus, NKR are extraordinarily rapidly evolving receptors that must adapt to rapidly changing pathogens. Therefore we were surprised to identify the activating NKR, NKp30 in the amphibian Xenopus and cartilaginous fish (shark, skate) databases. NKp30 is MHC-linked in humans and possesses a unique V-type immunoglobulin superfamily (IgSF) domain, the so-called “VJ”-type, which resembles the precursor of antigen receptors (AgR) and is assumed to have predated the emergence of AgR. Further database searches for NKR in jawless vertebrates and lower deuterostomes revealed no NKR orthologs, implying that NKp30 is the most conserved and oldest NKR in jawed vertebrates. While mammalian NKp30 is encoded by a single copy gene, with a cytoplasmic region that interacts with an ITAM-containing adaptor molecule, multiple NKp30 genes were found in Xenopus and sharks, some with ITIM-containing cytoplasmic regions, indicating inhibitory activity characteristic of other NKRs like KIR/CD94-NKG2. We are testing whether the MHC association of NKp30 is primordial, which would further suggest its coevolution with other MHC-linked proteins such as class I/II or B7 family members.
Comparative cancer immunology: Inducing myeloid derived suppressor cells in vitro (P6092)
Authors: Papenfuss Tracey , VanGundy Zachary , Sherger Matthew , Wasserman Jacob 
Published: January 2023

Abstract
Comparative oncology has broad applications in the development and optimization of novel therapeutics for cancer in both humans and animals. In this study, we developed in vitro tools to study canine myeloid derived suppressor cells (MDSCs). Previously, we have shown that tumor derived soluble factors (TDSFs) modified the function of a dendritic cell line. In this study, we investigated TDSF’s ability to induce MDSCs. TDSFs were generated from canine tumor cell lines and canine bone marrow precursors were differentiated in the presence of GM-CSF with or without TDSFs. We found that TDSFs dramatically increased the expression of CD11b+CADO48A+ (a canine MDSC marker akin to CD11b+Gr1+ MDSCs in mice) in cells between days 3-7 compared to controls with the highest percentage (40%) occurring on day 5 of differentiation. Day 5 TDSF-exposed cells suppressed the proliferation of responder immune cells. Similar results were found in companion studies using immunocompetent mice and murine in vitro TDSF-generated MDSCs. Taken together, these findings support that TDSFs can generate canine MDSCs in vitro that are similar to mouse MDSCs and that the in vitro generation of MDSCs mimics the immunosuppression seen with ex vivo derived MDSCs in humans and mice. These data have important implications both in understanding canine cancer immunology and for translational and therapeutic applications utilizing the dog as a model for human cancer.
Identifying the cellular interactions required for diversification of the primary antibody repertoire in rabbit GALT (P6101)
Authors: Lanning Dennis , Knight Katherine 
Published: January 2023

Abstract
Rabbits generate an initial antibody repertoire of limited diversity due to preferential use of the 3’-most VH gene segment during V-D-J gene rearrangement. Around one week of age, rabbit B cells begin diversifying this antibody repertoire in gut-associated lymphoid tissue (GALT), by mutating their V-D-J genes in response to antigen-independent signals acquired from intestinal commensals. To identify the cell-cell and cell-microbe interactions required for V-D-J gene diversification, we examined B cell expression levels of four chemokine receptors that mediate homing to important sites within GALT. We stained the chemokine receptors CXCR4, CCR6, CXCR5 and CCR7 on appendix B cells from two- to eight-day-old rabbits with fluorescence-labeled antibodies or chemokine-Ig fusion proteins and analyzed them by flow cytometry. The changes in chemokine receptor expression we observed during the first week of life suggest that, after entering appendix follicles, B cells first home to the follicle-associated epithelium, where they likely encounter commensal bacteria sampled from the intestinal microbiota. B cells then home sequentially to the follicular dendritic cell network, the T cell area and finally to the basal region of the follicle, where they proliferate and diversify their V-D-J genes to generate the primary antibody repertoire.