bluuebunny/crossref_metadata_2025_split huggingface dataset:
Notch modulates human NK cell differentiation through regulation of inhibitory killer immunoglobulin-like receptor (KIR) expression and acquisition of effector function (TUM2P.890)
Authors: Felices Martin , Ankarlo Dave , Lenvik Todd , Miller Jeffrey 
Published: January 2023

Abstract
The Notch signaling pathway plays a substantial role on human NK cell development, however the role of Notch on KIR upregulation and acquisition of effector function has not been explored. To evaluate how Notch influences terminal differentiation, cord blood derived NK cells or sorted KIR- peripheral blood NK cells were cultured with IL-15 for 7 days in the presence or absence of gamma-secretase inhibitor, known to inhibit Notch signaling. Inhibition of Notch signaling significantly decreased KIR expression. Conversely, co-culturing the same cells with OP9 cells bearing Notch ligands enhanced KIR expression. Overexpression of the active portion of Notch on cord blood derived NK cells after 28 days of culture resulted in a 2-fold increase in KIR expression indicating that Notch signaling plays a direct, cell intrinsic, role in KIR regulation. By knocking down delta-like 1 (DLL1) on NK cells and co-culturing them with OP9 cells expressing Notch ligands we show that DLL1 mediated cis-inhibition controls KIR expression. Notch signaling also enhances CD16 upregulation that precedes KIR expression. More important than receptor expression is function. We found that Notch signaling induces increased cytolytic effector capacity and cytokine secretion, even on post-transplant samples where NK cell function is inherently defective. Given these attributes of Notch signaling, we propose Notch agonists can enhance NK cell maturation and tumor killing in a post-transplant setting.
Inhibition of tumor growth by targeting the MDA5-IPS-1 pathway requires coordinated induction of cancer cell death and immune activation (TUM2P.891)
Authors: Yu Xiaofei , Fisher Paul , Wang Xiang-Yang 
Published: January 2023

Abstract
Melanoma differentiation-associated antigen 5 (MDA5), a cytosolic innate pattern recognition receptor for sensing double-stranded RNA, functions as a first line of defense against viral infection. Here we report that ectopic expression of MDA5, mediated by a replication incompetent adenovirus (Ad.Mda5), results in prostate cancer cell death and concomitant innate immune activation, e.g., production of type I interferons (IFNs). IFN-β promoter stimulator (IPS)-1, a downstream adaptor protein, is critical for MDA5-stimulated cancer cell apoptosis and type I IFN response. While MDA5-regulated IFN regulatory factor 3-IFN-β signaling cascade remains intact in nontransformed normal cells, these cells are less sensitive to MDA5-induced apoptosis compared with cancer cells. Intriguingly, intratumoral delivery of Ad.Mda5 preferentially remodels the tumor environment toward Th1 polarization, indicated by marked elevation of cytokine IFN-β, IL-12, and IFN-gamma. As a result, in situ overexpression of MDA5 leads to strong suppression of established mouse prostate tumors, which is associated with activation of natural killer (NK) cells and cytotoxic CD8+ T lymphocytes. Genetic ablation or antibody neutralization reveals that antitumor effect of MDA5 therapy depends on IPS1, IFN-β, and IL-12. Therefore, the intrinsic capabilities of ‘danger’ sensing MDA-5 in simultaneously promoting cancer cell death and immune augmentation may be exploited to develop novel cancer therapeutics.
Multifunctional CD26hi Th17 cells eradicate large human tumors (TUM2P.902)
Authors: Nelson Michelle , Bailey Stefanie , Huff Logan , Kundimi Sreenath , Paulos Chrystal 
Published: January 2023

Abstract
Human CD4+ T cells differentiate into multiple effector subsets, but their distinct roles in anti-tumor immunity remain elusive. CD4+ T cell subsets enriched from bulk human CD4+ T cells [Th1 (CXCR3+), Th2 (CCR4+), and Th17 (CCR6+ or CD26hi)] were stimulated with anti-CD3 beads bearing agonists to either CD28 or ICOS and were then engineered with a chimeric antigen receptor that recognizes human mesothelioma. In vitro, ICOS costimulation proved superior to CD28 for augmenting the function of human Th1, Th2, CCR6+ Th17 and CD26hi Th17 cells, as indicated by elevated production of IFN-γ, IL-4 and IL-17A, respectively. Moreover, CD26hi Th17 cells possessed strikingly enhanced polyfunctionality compared to Th1, Th2 or CCR6+ Th17 cells, as demonstrated by their heightened capacity to co-secrete IL-17A, IFN-γ, IL-22, IL-2, and TNF-α simultaneously. Compared to other enriched T cell subsets, a greater percentage of CD26hi Th17 cells exhibit an effector memory phenotype. In vivo, CD26hi Th17 cells more efficiently reconstituted immunodeficient hosts and persisted long-term. Furthermore, CD26hi Th17 cells possessed a superior ability to kill large human tumors (&gt;150mm2) when infused into mice compared to Th1, Th2 or CCR6+ Th17 cells. These results suggest that the generation of multifunctional, long-lived human Th17 populations could be instrumental to the design of novel, effective T cell-based cancer therapies.
Isocitrate dehydrogenase mutant glial cell resist natural killer cell cytotoxicity (TUM2P.906)
Authors: Zhang Xiaoran , Chang Yigang , Brancho Michael , Kohanbash Gary , Leo Madeline , Okada Hideho , Lotze Michael , Amankulor Nduka 
Published: January 2023

Abstract
Introduction: Mutations in IDH 1 and 2 are common in gliomas. Methylation-related epigenetic repression of gene expression is a feature of IDH mutant (IDHmut) tumors. Here, we demonstrate transcriptional repression of NKG2D activating ligands (NKG2DLs) in IDHmut tumors, and correlate this with decreased susceptibility to natural killer (NK) cell-mediated cytolysis in vitro Methods: Microarray: We compared IDHmut and IDHwt gene expression and NKG2DL mean methylation values in glioma patients using TCGA data. RT-PCR: NKG2DL expression was assayed using primers for NKG2D ligands. Cytotoxicity: Astrocytes were cultured with activated NK cells. Cytotoxicity was measured by flow cytometry. Results: mRNA expression values for NKG2DLs ULBP1 and ULBP3 were significantly reduced in IDHmut gliomas. Conversely, methylation values were significantly increased. NKG2DL hierarchical clustering of gene expression of shows genotype-based expression clusters. IDHmt astrocytes expressed a two to eight-fold reduction in the relative gene expression of NKG2DLs compared with IDHwt astrocytes. Cytotoxicity is significantly lower in IDHmut astrocytes/NK cell co-cultures compared with IDHmut/NK or U87/NK co-cultures. Conclusions: Our findings demonstrate evidence for the transcriptional repression of NKG2DLs in IDHmut gliomas in silico and in vitro. Reduced NKG2D levels in IDHmut cells correlates with impaired NK-mediated cytotoxicity and suggests a role for NK cell resistance in IDH mutant tumors.
Clonally expanded T cells from skin biopsies of patients with systemic sclerosis recognize DNA topoisomerase I and cytomegalovirus antigens (HUM3P.258)
Authors: Zacharakis Nikolaos , Oleszak Emilia , Gaughan John , Nanajian Anthony , Myers Allen , Tsygankov Alexander , Platsoucas Chris 
Published: January 2023

Abstract
Systemic sclerosis (SSc) is a chronic autoimmune disease, characterized by immune system dysregulation, uncontrolled deposition of collagen and microvascular damage in the skin and several internal organs. T cells have a key role in the initiation and propagation of SSc. Our laboratory has previously identified the presence of high proportions of identical α- and β-chains TCR transcripts in skin biopsies from patients with SSc of recent onset, demonstrating clonal expansion of T cells in response to one or more antigens. In order to identify these antigens, pairs of those in vivo clonally expanded α- and β-chain TCR transcripts from two patients with SSc (SSc-21 and SSc-22) were expressed in mutant TCR negative T cells, J.RT3-T3.5, generating a total of 52 T cell lines, including 10 controls. An intracellular Ca2+ mobilization assay was employed to examine whether these transduced Jurkat T cell lines recognize putative SSc antigens presented by autologous EBV-transformed B cell lines. The self-antigen, DNA topoisomerase I and CMV and parvovirus antigens were tested. Statistically significant intracellular Ca2+ mobilization was observed in response to 3 DNA topoisomerase I and 2 CMV peptides by 5 T cell lines expressing clonally expanded α- and β-chain TCR transcripts from patient SSc-21 and SSc-22. These results demonstrate that clonally expanded T cells recognizing DNA topoisomerase I or CMV antigens are present in skin biopsies of recent onset of patients with SSc.
Broad macrophage infiltration leads to a proinflammatory state in streptozotocin-induced diabetic mice (HUM3P.261)
Authors: Niu Shuo , Bian Zhen , Tremblay Alexandra 
Published: January 2023

Abstract
Our studies demonstrate significantly enhanced neutrophil (PMN) recruitment during zymosan-induced peritonitis in mice with strepotozocin (STZ)-induced diabetes. Cross-leukocyte adoptive transfer further reveals that, while the PMN response is potentiated under diabetes, the existence of a proinflammatory state in the peritoneum of diabetic mice, which is reflected by more than two-fold increases in peritoneal macrophages and significant elevations of IL-6, IL-1β, TNF-α, and the PMN-driven chemokine CXCL1 (KC/IL-8) upon zymosan challenge, plays a dominant role in exaggerating the peritonitis. Insulin treatment not only alleviates hyperglycemia, but also reduces macrophage infiltration in the peritoneum and eliminates the enhanced PMN recruitment during the peritonitis, confirming that such proinflammatory state is associated with hyperglycemia but not STZ. Strikingly, dissections of various tissues from diabetic mice reveal increased macrophages infiltration in additional essential organs including the liver and kidneys. Exposing diabetic mice to a sub-effective dose of adriamycin, a potential renal damage drug, leads to acute nephritis and proteinuria toward which insulin treatment precludes. To address how diabetes induces broad macrophage infiltration, we analyzed peripheral leukocytes and observed a significant increase in the circulating proinflammatory F4/80+Gr-1+ monocytes, which infiltrate tissues and trigger exaggerated responses upon inflammatory challenges.
Cardiometabolic abnormalities associate with an inflammatory cytokine profile in breast adipose tissue and plasma of obese African American women (HUM3P.264)
Authors: Denis Gerald , Strissel Katherine 
Published: January 2023

Abstract
‘Crown-like structures’ (CLS) of CD68+ macrophages in human subcutaneous, visceral and breast adipose depots associate with cardiometabolic and cancer risks. Rather than surgical fat biopsy, which is difficult for vulnerable humans, informative blood-based assays are preferred for medical decision making and risk assessment. HYPOTHESIS. Breast CLS and adipocyte parameters, as measures of metabolic health, meaningfully associate with pro-inflammatory imbalances of specific cytokines and adipokines in plasma of obese women. RESULTS. We obtained breast histological samples and matched plasma of 20 obese, unhealthy African American women in the Komen Tissue Bank and the Black Women’s Health Study, and compared to 20 age-, sex- and race-matched subjects who were obese but healthy; and to a matched cohort who were lean and healthy. We measured 41 cytokines and adipokines in plasma, and determined CLS and adipocyte size (a measure of metabolic health) in breast fat. We used clinical determinations of cardiometabolic status as independent variables and evaluated patterns of cytokine expression and tissue immunometabolism as dependent variables. We found that obese and healthy women clearly resolve from obese and unhealthy as well as from lean and healthy, and that a limited set of cytokines and adipokines are sufficient to define risk cross-sectionally. IMPACT. Immunometabolic profile of patient plasma may offer a robust tool to assist patient-provider discussion of risk.
IL-7Ralow memory CD8+ T cells are clonally anergic cells and defect in aerobic glycolysis pathway (HUM4P.274)
Authors: Sim Ji Hyun , Kim Jin-Hee , Seol Min A , Lee Sun-Kyung , Cho Bon-A , Ko Youngho , Oh Keunhee , Lee Dong-Sup , Kim Joon , Park Ae , Kang Insoo , Choi Kyungho , Kim Hang-Rae 
Published: January 2023

Abstract
We have found two subsets of CD8+ T cells expressing IL-7Rahigh and IL-7Ralow with different cell survival responses to IL-7 in peripheral blood of human. Treating IL-7Rαlow memory CD8+ T cells with anti-CD3/CD28 Abs did not produce IL-2, but comparably expressed IFN-γ and TNF-a. But the mechanism for this is unknown. These cells have reduced calcium flux and subsequent inhibited nuclear translocation of NF-AT1 upon TCR stimulation as well as PMA/ionomycin treatment, suggesting that IL-7Rαlow memory CD8+ T cells also have defected in distal-TCR signaling. Importantly, this anergic status was reversed by exogenous IL-2 stimulation, clearly distinct from adaptive tolerance. When TCR stimulation on IL-7Rαlow memory CD8+ T cells exhibited markedly decreased GATA3 and c-myc expression and increased NUAK1, which are key regulators in aerobic glycolysis, compared to those of IL-7Rαhigh memory CD8+ T cells. The expression of GATA3 and c-myc in IL-7Rαlow memory CD8+ T cells were restored after IL-2 stimulation. In conclusion, we firstly show that specific subset of human CD8+ T cells, IL-7Rαlow memory CD8+ T cells, are T-cell clonally anergy in vivo. We also demonstrate that GATA3-c-myc-NUAK1 system may control the maintenance of clonally anergic IL-7Rαlow memory CD8+ T cells in vivo and provides insight into human immunology. Acknowledgement: This research was supported by National Research Foundation of Korea (2014M3A7B4052194) and by the SNUH Research Fund (34-2013-0206)
Statin treatment suppresses IgE-mediated mast cell activation in vitro and in vivo (HYP4P.316)
Authors: Barnstein Brian , Kolawole Elizabeth , Oskeritzian Carole , Ryan John 
Published: January 2023

Abstract
Statin drugs are HMG-CoA reductase inhibitors used in the treatment of hypercholesterolemia and cardiovascular disease. They have also been shown to suppress the immune response. Given the critical role that mast cells play in allergic and inflammatory diseases, we assessed the effect of statin drugs on mast cell function. We demonstrate that Fluvastatin suppresses IgE-mediated degranulation and production of pro-inflammatory cytokines and chemokines, without altering IL-10 secretion. These effects were reversed by the addition of mevalonic acid, and mimicked by geranyl geranyl tranferase blockade, suggesting that Fluvastatin inhibits lipid modifications to small G proteins downstream of HMG CoA reductase. In vivo studies further showed that Fluvastatin suppressed IgE-mediated anaphylaxis. Interestingly, Fluvastatin-mediated suppression was completely absent in 129/SvJ mast cells, which showed a 2-fold induction of HMG CoA reductase upon statin treatment. Human mast cells from eight donors showed a range of responses to Fluvastatin, indicating that genetic background may influence statin-mediated suppression. These data demonstrate the possible use of statins, a widely-prescribed group of drugs, as a possible therapy for mast cell-associated disorders.
Transcriptional profiling of constitutive mast cells reveals a unique gene expression signature and highlights their heterogeneity across tissues (HYP4P.303)
Authors: Dwyer Daniel , Barrett Nora , Austen K. 
Published: January 2023

Abstract
Constitutive mast cells (MC) are long-lived sentinel cells residing in peripheral connective tissue near the microvasculature and nerve endings. Similar to basophils (Ba), MC express the high-affinity IgE receptor FcεR1α, and are frequently implicated in the pathobiology of allergic diseases. Although protease expression in murine MC varies between tissues, the full extent of their diversity across compartments is not known, and the study of MCs is hindered by their rarity and location. In collaboration with the Immunological Genome Project, we developed techniques for isolating MC from a wide variety of murine tissues for microarray analysis. Hierarchical clustering and principal component analysis of MC versus Ba and other immune cells indicate that Ba are transcriptionally more similar to other circulating granulocytes (eosinophils, and neutrophils) than to MC. Furthermore, granulocytes and other tissue resident myeloid cells (macrophages and dendritic cells) are closer to each other and to lymphoid cells than either group is to constitutive MC. Further analysis revealed a core MC transcriptional signature not seen in other immune cell populations, but MC heterogeneity across tissue compartments was similar to that seen with dendritic cells, indicating that their diversity goes far beyond protease expression. Our results provide novel insights into this unique cell type and lay the groundwork for future studies on the activation and function of MC in peripheral tissue.
Developing an antibody targeting CεmX of mIgE for the treatment of IgE-mediated diseases (HYP4P.305)
Authors: Chan Yueh-Hsuan , Lee Yu-Tzu , Chou Hung-Wen , Wu Pheidias , Chen Jiun-Bo , Li Chau-Hong , Cheng Tien-Tien , Chen Nien-Yi , Chang Tse-Wen , Lue Ko-Haung 
Published: January 2023

Abstract
CεmX (also referred to as M1’) is a discrete domain of 52 a.a. residues, located between the CH4 domain and the C-terminal membrane anchor peptide of the ε heavy chain of membrane-bound IgE (mIgE) on human B lymphocytes. Antibodies that target CεmX are potentially useful in controlling IgE production for treating allergic and other IgE-mediated diseases. Herein we report that an anti-CεmX mAb, 4B12, was shown to be effective in reducing allergen-specific IgE and IL-5 production upon the challenge of the allergen in an asthma model employing CεmX gene knocked-in mice that express mIgE containing human CεmX domain on B cells. 4B12 could also alleviate airway hyper-responsiveness (AHR), allergen-induced eosinophil infiltration, and lung inflammation in those mice. Furthermore, we demonstrated that a humanized 4B12 mAb (referred to as FB825) could inhibit the production of human IgE in mice that had been reconstituted with human peripheral blood mononuclear cells. Based on the abilities of 4B12 to bind to mIgE and to lyse mIgE-expressing B cells by apoptosis, ADCC, and other cytolytic mechanisms and on the above results on animal models, a phase-I human clinical trial of FB825 to investigate its IgE-related biological effects is being performed.
Unexpected pro-inflammatory effects of IL-10 on mouse mast cells (HYP4P.306)
Authors: Abdul Qayum Amina , Ryan John 
Published: January 2023

Abstract
Mast cells play critical roles in allergic and inflammatory diseases as effector and immunoregulatory cells. A well understood mast cell activation pathway is antigen-mediated crosslinkage of IgE bound to its high affinity receptor (FcεRI). Cytokines produced during the immune response may alter mast cell function. We have shown that, contrary to its better-known anti-inflammatory role, IL-10 induces pro-inflammatory cytokines in peritoneal and bone marrow derived mast cells stimulated with IgE+antigen. The increase in pro-inflammatory cytokines and chemokines (IL-6, MCP-1, MIP-1a, and IL-13) is seen both at the mRNA and protein level. Consistent with previously published data, IL-10 suppresses TNF production. Using an in vivo IgE-induced passive systemic anaphylaxis model, we show that IL-10 pre-treatment causes a more severe anaphylactic response and increased serum IL-6 and MIP-1a, suggesting that IL-10-mediated enhancement can occur in vivo. These data support a more nuanced view of IL-10 as a key immunoregulatory cytokine capable of curbing and promoting different inflammatory mediators in hypersensitivity reactions.
Identification of a novel T. cruzi B cell superantigen (INC4P.340)
Authors: Brown Eric 
Published: January 2023

Abstract
Catalytic antibodies represent an innate defense mechanism that allows a subset of antibodies to bind and then hydrolyze their targets, making these antibodies up 1000-fold more efficient than conventional immunoglobulins. Pathogens have evolved B-cell superantigens (BC-SAgs) as an immunevasion strategy designed to neutralize catalytic antibody activity. Exposure to BC-SAgs such as the Staphylococcus aureus extracellular fibrinogen binding protein (Efb) and the HIV gp120 glycoprotein decreased catalytic activity to these antigens, respectively. We describe for the first time the identification and characterization of a Trypanosoma cruzi BC-SAg (Tc24). Immunization with Tc24 in the presence of different adjuvants significantly decreased the ability of purified IgM to hydrolyze Tc24 compared hydrolysis rates observed for IgM purified from unimmunized controls. Infection with T. cruzi resulted in a similar decrease in catalytic activity. In both immunization and infection models, the dose (of either Tc24 or the number of T. cruzi used to infect mice) affected Tc24 hydrolysis rates. Analysis of IgMs purified from infected humans also demonstrated reduced hydrolysis of Tc24 compared to hydrolysis rates observed for IgM purified from uninfected individuals. These data suggest that Tc24 is a BC-SAg that may impact the host's defenses early in the infection process.
Macrophages modulate bacterial persistence and gastric pathology during Helicobacter pylori infection (INC4P.346)
Authors: Viladomiu Monica , Bassaganya-Riera Josep , Kronsteiner-Dobramysl Barbara , Washington Philipson Casandra , Michalak Pawel , Eden Kristin , Hontecillas Raquel 
Published: January 2023

Abstract
Based on preliminary data suggesting that macrophages are critical regulators of Helicobacter pylori colonization and gastric pathology, we investigated how macrophage phenotypes shape the outcome of infection and persistence. The loss of PPARγ in the myeloid compartment favors a 5 to 10-fold decrease in bacterial loads at the expense of more severe gastric lesions. PPARγ-deficient macrophages present altered control of transcription factors such as NFκB, which results in a pro-inflammatory phenotype. Stomach lamina propria analyses at weeks 0 to 4 post-infection revealed increased levels of CD11b+ F4/80hi CD64+ CR3CR1+ macrophages in WT mice, while PPARγKO mice failed to expand and maintain such population. Macrophage depletion using clodronate liposomes resulted in a significant reduction of gastric H. pylori in WT mice, thus abrogating the differences in bacterial loads observed between WT and PPARγKO mice. In vitro co-culture of H. pylori with bone marrow derived macrophages showed that either PPARγ loss or pharmacological blockade enhances bactericidal activity, which is associated to significant differential expression of chil1, etv5, iigp1, ptger4, sqle, osm, hspa2 and rptoros levels as revealed by global transcriptome analyses. Hence, macrophages facilitate H. pylori infection by 1) serving as bacterial reservoirs and allowing intracellular replication, and 2) favoring a gastric regulatory response that favors persistence.