bluuebunny/crossref_metadata_2025_split huggingface dataset:
MIF contributes to Type 1 diabetes favors insulitis by mediating macrophages and dendritic cells maturation (P4533)
Authors: Sanchez Zamora Yuriko , Rodriguez Sosa Miriam 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is characterized by a cellular infiltrate in pancreatic islets where β cells are destroyed. The recognition of antigens and auto antigens takes place by macrophages (Mo) and dendritic cells (DCs). Previously we have showed that MIF favors the expression of co-stimulatory molecules on Mo and DCs on infection diseases. However, the role of MIF on Mo and CDs in T1D has not been explored. Here, we determined the expression of co-stimulatory molecules on Mo and CDs from WT or MIF-/- mice on experimental T1D induced by STZ. Cells extracted from pancreas and spleens were treated with antibodies anti- CD80, CD86, CD40, MHC-II and TLRs. Also we tested the number of CD4+CD25+FOXp3+ cells. The results show that MIF-/- mice not developed high glucose levels compared with WT mice. Pro-inflammatory cytokines in serum were diminished on MIF-/-STZ mice, while the anti-inflammatory cytokines were higher compared to WT STZ mice. Moreover, MIF-/-STZ mice had less CD80, CD86, MHCII, TLR-2 and TLR-4 in spleen and pancreatic islets Mo and CDs. In addition, CD4+CD25+ spleen cells had a high expression of FOXp3 transcription factor in MIF-/- healthy than MIF+/+ healthy mice. 8 weeks after of STZ induction the MIF-/-STZ mice present higher FOXp3 expression than WT STZ mice. Our results suggest that MIF favors the expression of co-stimulatory molecules in the Mo and CDs in T1D. Additionally we propose that MIF down-regulate the proliferation of regulatory T cells in T1D.
Determination of anti-PF4/heparin complex antibodies in the fluid phase (P4549)
Authors: Schulze Annika , Alahmad Adnan , Krauel Krystin , Jensch Inga , Greinacher Andreas , Hundt Matthias 
Published: January 2023

Abstract
Antibodies against the human platelet factor 4/heparin complex (PF4/H) are found in the immune-mediated adverse drug reaction heparin-induced thrombocytopenia. These antibodies are directed against PF4 within the complex due to conformational changes of PF4. To study the humoral immune response to PF4/H in greater detail, C57BL/6 mice were immunized with murine (mu) PF4/H by retro-orbital injection. Serum was obtained, and analyzed by ELISA either with muPF4/H directly plate-bound (solid phase), or in a fluid phase assay with muPF4/H that contained 10% biotinylated muPF4. The two approaches differed notably in their specificity and sensitivity to detect anti-muPF4/H antibodies. The solid phase ELISA could not distinguish between antibodies against plate-bound muPF4 alone and plate-bound muPF4/H. Only incubation with excess heparin allowed to distinguish these antibodies. In contrast, the fluid phase ELISA permitted the direct distinction between anti-muPF4 and anti-muPF4/H antibodies. The fluid phase system was also successfully applied to detect muPF4/H-specific B cells by ELISpot. Thus, the fluid phase ELISA for the detection of anti-muPF4/H antibodies was superior to the solid phase assay. Furthermore, our data suggest that surface binding of PF4 might induce conformational changes similar to those induced by heparin binding. Finally, fluid phase ELISAs might be preferable for the detection of antibodies that bind to conformation-dependent epitopes.
Serum amyloid A stimulates macrophage foam cell formation via upregulation of lectin-like oxidized low-density lipoprotein receptor 1 (P5188)
Authors: Jung Young Su , Lee Ha Young , Lee Sung Kyun , Kim Sang Doo , Baek Suk-Hwan , Bae Yoe-Sik 
Published: January 2023

Abstract
Elevated levels of serum amyloid A (SAA) is a risk factor for cardiovascular diseases, however, the role of SAA in the pathophysiology of atherosclerosis remains unclear. Here we show that SAA induced macrophage foam cell formation. SAA-stimulated foam cell formation was mediated by c-jun N-terminal kinase (JNK) signaling. Moreover, both SAA and SAA-conjugated high density lipoprotein stimulated the expression of the important scavenger receptor lectin-like oxidized low-density lipoprotein receptor 1 (LOX1) via nuclear factor-kB (NF-kB). A LOX1 antagonist carrageenan significantly blocked SAA-induced foam cell formation, indicating that SAA promotes foam cell formation via LOX1 expression. Our findings therefore suggest that SAA stimulates foam cell formation via LOX1 induction, and thus likely contributes to atherogenesis.
The tandem PH domain-containing protein 2 regulates chemokine-induced cytoskeletal reorganization and malignant B cell migration (P5194)
Authors: Li Hongzhao , Hou Sen , Wu Xun , Nandagopal Saravanan , Lin Francis , Kung Sam , Marshall Aaron 
Published: January 2023

Abstract
The tandem PH domain-containing protein 2 (TAPP2), along with its homologue TAPP1, is best known for high-specificity binding to PI(3,4)P2, a lipid product of phosphoinositide 3-kinases (PI3K). While PI3K enzymes have a number of functions in cell biology, including cell migration, the functions of PI(3,4)P2 and its binding proteins are not well understood. Previously we found that TAPP2 is highly expressed in leukemic B cells with strong migratory capacity. Here we show that the migration of human malignant B cells is significantly inhibited by TAPP2 knockdown (KD), manifested as reduction in percentage of migrating cells, migration velocity and directionality. TAPP2 KD led to alterations in chemokine-induced rearrangement of the actin cytoskeleton and failure to form polarized morphology. TAPP2 co-localized with the stable F-actin-binding protein utrophin, with both molecules reciprocally localizing against F-actin accumulated at the leading edge. In TAPP2 KD cells, Rac was over-activated and localized to multiple membrane protrusions, suggesting that TAPP2 may act in concert with utrophin and stable F-actin to spatially restrict Rac activation and reduce formation of multiple membrane protrusions. In summary, this study identified TAPP2 as a novel regulator of malignant B cell migration and a potential therapeutic intervention target.
ER stress and neuroinflammation: connecting the unfolded protein response to JAK/STAT signaling (P5196)
Authors: Meares Gordon , Benveniste Etty 
Published: January 2023

Abstract
Neuroinflammation and endoplasmic reticulum (ER) stress are associated with many neurological diseases. ER stress is brought on by misfolded proteins. In turn, cells respond with activation of the unfolded protein response (UPR). The UPR is a highly conserved pathway that transmits both adaptive and apoptotic signals to restore homeostasis or eliminate the irreparably damaged cell. Recent evidence indicates that ER stress and inflammation are linked. In this study, we have examined the interaction between ER stress and JAK/STAT-dependent inflammation in astrocytes. The JAK/STAT pathway mediates the biological actions of many cytokines and growth factors. We have found that ER stress leads to the activation of STAT3 in a JAK1-dependent fashion. ER stress-induced activation of the JAK1/STAT3 axis leads to expression of IL-6 and several chemokines. The activation of STAT3 signaling is dependent on the protein kinase PERK, a central component of the UPR. Knockdown of PERK abrogates ER stress-induced activation of STAT3 and overexpression of PERK is sufficient to activate STAT3. Additionally, ER stressed astrocytes, via paracrine signaling, can stimulate activation of microglia leading to production of oncostatin M (OSM). OSM can then synergize with ER stress in astrocytes to drive inflammation. Together, this work describes a new PERK-JAK1-STAT3 signaling pathway that may elicit a feed-forward inflammatory loop involving astrocytes and microglia to drive neuroinflammation.
Host macrophages are a barrier to the engraftment of embryonic stem cell-derived hematopoietic progenitors. (P4544)
Authors: Thompson Heather , McLelland Bryce , van Rooijen Nico , Manilay Jennifer 
Published: January 2023

Abstract
Embryonic stem cell-derived hematopoietic progenitors (ES-HPs) have been proposed as an alternative bone marrow (BM) source. ES-HPs display features of adult hematopoietic progenitors, but do not achieve high levels of engraftment after transplantation, even in immunocompromised hosts. We hypothesized that host macrophages (Mϕ) are a barrier to ES-HP engraftment. To test this, we cultured and transplanted CD41+ ES-HPs into sublethally irradiated NOD.SCID.γc (NSG) mice that lack B, T and NK cells, but still possess Mϕ. Less than 5% ES-HP donor chimerism in the BM and spleen was observed in NSG recipients, in contrast to &gt;90% chimerism in adult BM-transplanted controls. In addition, we noticed a 3.5-fold increase in spleen weight and a 1.75-fold increase in splenic host F4/80+ Mϕ in ES-HP recipients. In vitro phagocytosis assays confirmed that allogeneic Mϕ phagocytosed ES-HPs at higher levels compared to adult Lin- BM targets. Interestingly, syngeneic Mϕ also engulfed ES-HPs, but not adult Lin- BM. When Mϕ were eliminated with liposome-encapsulated clodronate, a 2.17-fold increase in ES-HP engraftment in the BM and 5.14-fold increase in the spleen was observed. Taken together, our data demonstrate that F4/80+ Mϕ are a barrier to ES-HP engraftment and suggest that cultured ES-HPs are unable to regulate host Mϕ phagocytosis. In line with this, ES-HPs expressed low levels of the Mϕ inhibitory ligand, CD47. Functional studies using CD47-Fc are in progress.
Antibodies against ROS-producing ragweed pollen proteins may prevent allergic sensitization (P6375)
Authors: Pazmandi Kitti , Kumar Brahma , Boldogh Istvan , Kemeny-Beke Adam , Szilasi Maria , Rajnavolgyi Eva , Bacsi Attila 
Published: January 2023

Abstract
We have previously described that hydrated pollen grains and their allergenic extracts possess intrinsic NAD(P)H oxidase activity, which catalyzes the production of reactive oxygen species (ROS) and induces oxidative stress in the lung or conjunctiva. This oxidative insult promotes the development of adaptive immune responses against innocuous pollen proteins and augments allergic airway inflammation induced by pollen antigens. In this study we investigated the ROS-producing activity of short ragweed (Ambrosia artemisiifolia) pollen extract (RWE) in detail. First, we found that in the presence of NADPH, RWE produces more ROS than after addition of NADH. An in situ nitroblue tetrazolium assay demonstrated that at least two distinct enzymes are responsible for ROS generation in RWE. The first one can use both NADPH and NADH as substrates, while the second one produces ROS only in the presence of NADPH. We observed that despite the fact that sera from non-allergic individuals have lower non-enzymatic antioxidant capacity, they exhibit higher efficacy in inhibition of ROS generation by RWE compared to sera from allergic patients. Western blot analysis revealed that antibodies against some pollen proteins associated with ROS production of RWE are present only in the sera of non-allergic subjects. Our novel findings raise the possibility that these antibodies may be able to block the ROS production by pollen enzymes hereby prevent the sensitization of non-allergic individuals.
Helicobacter pylori-derived heat shock protein 60 increases the induction of regulatory T cells (P3365)
Authors: Hsu Wei-Tung , Wu Ming-Shiang , Wu Chang-Jer , Chen Chia-Hung , Jian Ting-Yan , Liao Kuang-Wen 
Published: January 2023

Abstract
Regulatory T cells have been shown to highly infiltrate the H. pylori-infected gastric mucosa. Treg cells suppress the host’s immune responses to H. pylori and dominate during persistent infection. H. pylori-derived HSP60 is one important virulence factor that promotes malignant tumorigenesis. This study further investigated the role of HpHSP60 in immunosuppression, and its activity was found to correlate with the induction of Treg cells. To determine the effects of HpHSP60 on immunosuppression, human peripheral blood mononuclear cells were treated with or without HpHSP60 in the presence of anti-CD3 mAb to determine the effect of HpHSP60 on proliferation. We subsequently asked whether proliferative inhibition correlates with the induction of Treg cells. Finally, the mechanism through which HpHSP60 induces Treg cells was identified. HpHSP60 decreased the expression of CDK4 to significantly slow the proliferation of mitogen-stimulated T cells, which correlated with the induction of Treg cells. Moreover, secretion of IL-10 and TGF-β by monocytic cells was essential to the induction of HpHSP60-induced Treg cells. In summary, we propose that HpHSP60 can act on macrophages to trigger the expression of IL-10 and TGF-β, resulting in an increase in Treg cells and the proliferative inhibition of T cells.
Analysis of bronchoalveolar immune cells and cytokines induced by immunization with different respiratory syncytial virus vaccines (P4395)
Authors: Ko Eun-Ju , Lee Jong Seok , Lee Yu-Na , Kwon Young-Man , Hwang Hye Suk , Yoo Si-Eun , Quan Fu-Shi , Lee Sujin , Moore Martin , Kang Sang-Moo 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infection and deaths in infants. Since the failure of formalin-inactivated RSV (FI-RSV) due to vaccine-enhanced pulmonary eosinophilia and pneumonia during 1960s, there is no licensed vaccine against RSV. In this study, we analyzed immune cell phenotypes and cytokines in bronchoalveolar lavage fluids (BALF) from mice immunized with FI-RSV, live RSV, or F plasmid and virus-like particles (VLP) containing RSV F and G glycoproteins (FFG VLP) at day 5 post RSV challenge. FI-RSV immunized mice showed severe eosinophilia, high levels of CD11b+ cells as well as IFN-gamma and IL-4 secreting CD4 T cells locally and systemically. Mice pre-immunized with live RSV showed no severe pulmonary eosinophilia, similar levels of IFN-gamma secreting CD4 and CD8 T cells, and low levels of IL-4 secreting cells. Mice immunized with FFG VLPs displayed no eosinophilia, relatively a low ratio of CD4/CD8 T cells secreting IFN-gamma, and no systemic cytokines. All 3 types of RSV vaccines were effective in clearing lung viral loads. Taken together, these results suggest no correlation between lung RSV clearance and inflammatory pulmonary disease, and that FFG VLP can be a promising RSV vaccine candidate conferring protection without lung disease.
Scavenger receptor SRECI can bind to Toll like receptor 4 and activate LPS-mediated signaling (P1362)
Authors: Murshid Ayesha , Gong Jianlin , Calderwood Stuart 
Published: January 2023

Abstract
Transmembrane receptor TLR4 was shown to induce the inflammatory response in the presence of its ligand, LPS. All TLRs belong to a family of PRR (pattern recognition receptor) which recognize PAMPs (pathogen-associated molecular patterns). The other receptor families, scavenger receptors work along with TLRs to attenuate innate immunity. SREC-I is a scavenger receptor expressed by immune cells and has a role in CD8+ T cell immunity. SREC-I can also modulate the function of Toll like receptors which have essential roles in innate immunity. Here we show that SREC-I can activate LPS mediated NFkB signaling. SREC-I can bind to TLR4 in the presence of LPS. We found that human monocyte/macrophage THP1 cells as well as bone marrow derived macrophages from mice exhibited increased responses to LPS and enhanced production of inflammatory cytokines such as IL-8, IL-6 with overexpressed SREC-I. We were able to show that downstream activation of NFkB requires either MyD88 or Trif adaptor protein. Our data suggested that SRECI is a cell surface receptor capable of internalizing LPS along with TLR4 as well as activating TLR4 mediated downstream cytokine production in macrophages.
Hyperglycemia triggers epigenetic machinery to enhance MyD88 expression in macrophages from type 1 diabetes mice (P1350)
Authors: Filgueiras Luciano , Sisti Flavia , Ferreira Ana , Wang Soujuan , Wang Zhuo , Jancar Sonia , Serezani C. Henrique 
Published: January 2023

Abstract
Type I diabetes (T1DM) is characterized by hyperglycemia and insulin deficiency. T1DM is associated with constitutive production of pro-inflammatory cytokine such as the members of IL-1 family. IL-1β binding to its MyD88-associated receptor IL-1R is a major cause of tissue injury in T1DM. We hypothesize that hyperglycemia (HG) enhances MyD88 in macrophages (MA) amplifying IL-1βR responsiveness. We found that both in vitro and in vivo HG increases the expression of MyD88 and its transcription factor STAT-1. Speculating that enhanced MyD88/STAT-1 levels could be due to epigenetic changes, we analyzed the expression of enzymes involved in DNA methylation (DNMT) and Histone deacetylation (HDAC) in MA from T1DM mice. We observed that diabetic mice expressed at least 2 times more HDAC 8, 10 and 11, and DNMT 3b than nondiabetic mice. In vitro inhibition of HDAC (TSA) or DNMT (5-azacyd) in MA impaired HG-enhanced MyD88 and STAT-1 expression. These data suggest a heretofore unrecognized epigenetic changes associated with hyperinflammatory pattern of MA from T1DM, leading to high MyD88 expression and possibly allowing overly IL-1β receptor and other MyD88-dependent receptor activation
Reactive oxygen probes - a broad range of colors with easier labeling and compatibility with fixation: novel CellROX® reagents from Molecular Probes® (P3295)
Authors: Bone Bradley , Seredick Barbara , Olszowy Mike 
Published: January 2023

Abstract
A natural consequence of aerobic respiration is the generation of highly toxic radicals called reactive oxygen species (ROS). ROS have been implicated in various human pathologies. Although fluorogenic probes such as Aminophenyl fluorescein (APF) and hydroxyphenyl fluorescein (HPF), and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) have been widely used to detect ROS using flow cytometry and imaging platforms, all are excitable by the 488nm laser and emit in the fluorescein channel, and require that labeling occur in a protein-free buffer. In contrast, the novel CellROX® ROS detection reagents from Molecular Probes® offer increased flexibility for multiplex experiments, with fluorescence emission in the fluorescein, PE, or APC channels for CellROX® Green, Orange, and Deep Red. In addition, the CellROX® reagents offer increased ease of use with the ability to label cells in complete growth media, have increased photostability as compared to H2DCFDA, and are compatible with fixation (CellROX® Green and Deep Red). The CellROX® ROS detection reagents are bright and stable ROS sensors that offer significant advantages over existing ROS sensors because they are compatible with labeling in different media and can be used with fixatives. For Research Use Only. Not for use in diagnostic procedures.
A single laser flow cytometer: simplifying multi-color immunophenotyping. (P3303)
Authors: Rohrer Jurg , Duckett Larry , Gaylord Brent , Stall Alan , Nguyen Andrea , Vidal Jason 
Published: January 2023

Abstract
Flow cytometry has been an indispensable tool for immunologists, clinicians and cell biologists for almost 40 years. The power of flow cytometry is in its ability to simultaneously measure multiple features (parameters) per cell and this unique aspect has formed the basis for most cell analysis applications to date. Initially cytometers were only able to distinguish 2 fluorescent dyes (or colors) but the technology has since advanced to multi-laser systems capable of up to 18 color detection. However, these instruments can be considered complex and costly for simple immunophenotyping experiments. The BD Accuri C6 was able to address this need as a simple to use instrument that could perform 4 color immunophenotyping on an individual’s lab bench as opposed to relying on core facilities. We have taken this concept a step further leveraging the exceptionally bright polymeric Sirigen dyes. These dyes, excited off the violet laser, span the visible to near IR spectrum giving one the opportunity to perform up to four color immunophentyping with minimal compensation while maintaining good signal resolution. In support of this, we will show data for simple T cell, B cell and NK cell immunophenotyping using a customized single violet laser Accuri. Details on optimizing filter sets will be given as well as details on how to optimize the reagent panels to be used with this laser/filter set up.
Apoptotic cell recognition receptors, Tyro3, Axl and Mer, demonstrate distinct patterns of ligand-inducible and ligand-independent receptor activation (P2016)
Authors: Tsou Wen-I , Nguyen Khanh Quynh , Birge Raymond , Kotenko Sergei 
Published: January 2023

Abstract
Proper recognition and removal of apoptotic cells by phagocytes lead to the active suppression of inflammatory responses and the induction of tolerance. Tyro3, Axl and Mer (TAM) receptor tyrosine kinases recognize apoptotic cells through their ligands, Protein S (ProS) and Growth-Arrest-Specific Gene 6 (GAS6), which in turn interact with phosphatidylserine (PS) exposed on the surface of apoptotic cells. Although TAMs share significant similarity, very little is known about the specificity of interaction between TAMs and their ligands, in the context of apoptotic cells, and about downstream signaling cascades triggered through TAMs. To study ligand-receptor interaction, we generated a series of reporter cell lines expressing chimeric TAM receptors that allowed us to demonstrate that each TAM has a unique pattern of interaction with GAS6 and ProS, which is also differentially affected by the presence of apoptotic cells. We also developed another set of chimeric receptors that triggered ligand-independent activation of the TAM intracellular domains, leading to distinct patterns of phosphorylated signaling molecules, as well as different levels of cytokine induction. Overall, these studies suggest that despite their similarity, Tyro3, Axl and Mer are functionally unique in the recognition of apoptotic cells, activation of downstream signaling cascades and gene expression, and therefore, contribute differently to cell survival and the induction of tolerance during tumorigenesis.
Antibodies of a novel bunyavirus in domestic and captive farmed animals in the Midwestern United States (P6371)
Authors: Xing Zheng , Schefers Jeremy , Schwabenlander Marc , Jiao Yongjun , Liang Mifang , Qi Xian , Li Chuan , Goyal Sagar , Cardona Carol , Li Dexin , Collins James , Murtaugh Michael 
Published: January 2023

Abstract
The Heartland virus (HLV), a new phlebovirus isolated in Missouri from patients with a history of tick bites and symptoms similar to severe fever with thrombocytopenia syndrome (SFTS), is an emerging pathogen. Genetically HLV is closely related to the newly identified bunyavirus (SFTSV) causing SFTS. In this study we tried to identify animal hosts that may be susceptible to and infected with HLV, or a SFTSV-like virus, and may serve as amplifying hosts important in virus transmission in the United States. Blood samples were obtained from several domestic and captive farmed animal species from 29 counties of Minnesota in 2012. An antigen sandwich ELISA for detection of SFTSV NP antibodies was used. Antibodies to SFTSV NP were detected in sera from 64 of 414 cattle (15.5%), 10 of 92 goats (10.9%), 6 of 48 sheep (12.5%), 35 of 296 white-tailed deer (11.8%), and 7 of 39 elk (18.0%). Thirty-four of 64 positive samples had P/N ratios between 4 and 10; and 11 had ratios ≥10. Our data show that both domestic and captive farmed animals were exposed to SFTSV- or HLV-like virus in the Midwestern United States. Since SFTSV and HLV are closely related, the viruses detected in the Midwestern region are most likely HLV or close relatives of HLV. Evidence that a novel phlebovirus infects a variety of domesticated and captive farmed animals as shown in this study validates the concern that SFTSV or HLV could pose a serious threat to animal welfare and public health in the United States.