bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cancer-induced activation of CD11b+Gr-1+ cells predispose hosts to antibody-independent anaphylactoid-type reactions (36.5)
Authors: LaFace Drake M , Pande Kalyan , Sathe Manjiri , McClanahan Terrill K , Phillips Joseph H 
Published: January 2023

Abstract
Cancer can induce expansion of CD11b+Gr-1+ myeloid cells, attributed to immune suppression. We noted tumor-activation of these cells further correlated with predisposition for anaphylactoid-type reactions in mice bearing 4T1 mammary carcinomas, which induces strong CD11b+Gr-1+ expansion. To test this hypothesis, 4T1-tumor bearing (4T1+) mice were challenged with recombinant adenoviral vector (rAd), previously shown to activate myeloid lineage cells. When challenged with a single intravenous dose of rAd, 4T1+ mice succumbed to acute anaphylactoid-type reactions. In contrast, minimal responses were seen in rAd treated mice implanted with the related cell line, 66cl4, which do not induce CD11b+Gr-1+ cells. Liposome depletion prior to rAd challenge protected 4T1+ mice, whereas, passive transfer of CD11b+Gr-1+ cells from 4T1+ mice was sufficient to convey susceptibility for anaphylactoid reactions in normal mice. Metabolic pathways were upregulated for nitric oxide and leukotrienes in the 4T1-induced CD11b+Gr-1+ cells. Pre-treating 4T1+ mice with inhibitors of NO-synthetase and leukotrienes inhibited rAd-induced anaphylaxis. Thus, malignant tumor growth can alter CD11b+Gr-1+ myeloid cells, rendering hosts susceptible to anaphylactoid-type reactions. Importantly, there was no requirement for pre-existing Ag-specific IgE or IgG. This reflects clinical reports in which cancer patients undergo treatment-induced anaphylactoid responses in absence of pre-exposure and may provide insight for modeling alternative pathways induced by direct activation of inflammation- or tumor-modified myeloid cells.
* Funded by the Schering-Plough Research Institute
Pattern of skin prick test reactivity to aeroallergens in patients with asthma and rhinitis residing in Riyadh, Saudi Arabia. (36.6)
Authors: Almogren Adel 
Published: January 2023

Abstract
The aim of this study was to determine the pattern of skin prick test reactivity to aeroallergens in patients with asthma and rhinitis (airway allergy) residing in Riyadh region.
During the period between Jan. 2003 and March 2004, allergy skin prick test results of 139 Saudi nationals (53% females and 47% males, mean age 27 + 12 years) with a clinical diagnosis of airway allergy residing in Riyadh region were analyzed in retrospect at King Khalid University Hospital Riyadh. A set of aeroallergens extracts for both indoor and outdoor allergens including fungal spores was used to test the patients.
Of the total 75% (105) patients with airway allergy reacted to one or more allergen extracts. Most frequently reacting indoor allergen was house dust mite (77.8%) followed by the cat (33.6%) and cockroach (19.2%) allergens. Among the outdoor allergens Prosopis juliflora tested positive in 72.1%, Bermuda grass in 53.8%, Chenopodium album in 47.1%, Rye grass in 36.5% and Salsola kali in 36.5%. A significant proportion of patients were also found reacting to Moulds (18.2%) and Aspergillus fumigatus (18.2%) extracts.
Sensitivity to one or more aeroallergens was common in atopic patients, indicating high level of aeroallergen sensitization in patients with airway allergy residing in Riyadh region.
Exploring Stem Cells: the Role of Stem Cells in Immune System Function (Educational Unit) (37.1)
Authors: Thoune Monica 
Published: January 2023

Abstract
Exploring Stem Cells: The Role of Stem Cells in Immune System Function is an educational unit developed for high school life science courses. Students will learn about embryonic and adult stem cells and the role adult stem cells play in the Immune System's function.
Students will learn about the tendency of Systemic Lupus Erythematosus (SLE) and Rheumatoid Arthritis (RA) patients to develop premature cardiovascular disease, specifically atherosclerosis. The unit will explore how the body's immune system, through the production of interferon αmay interfere with the normal development of endothelial progenitor cells (EPCs). Interferons are cytokines that are involved in immune system regulation and the inflammatory response. EPCs are bone marrow-derived stem cells which circulate in the bloodstream and participate in endothelial repair of the lining of blood vessels of the cardiovascular system.
This 5-6 day unit includes pedagogical strategies such as Internet treasure hunts, online interactive tutorials, cooperative group activities, problem solving, and hands-on activities. Students will gain real-world laboratory experience by working with micropipettes during a hands-on activity. They will learn how following proper protocol with minute samples is essential for accuracy and precision.
G-Protein coupled receptor kinases mediate TNF alpha-induced NF kappaB signaling via direct interaction with and phosphorylation of I kappa B alpha (38.15)
Authors: Patial Sonika , Luo Jiansong , Porter Katie Johanna , Benovic Jeffrey L , Parameswaran Narayanan 
Published: January 2023

Abstract
Tumor necrosis factor-α (TNFα) is a multifunctional cytokine involved in the pathophysiology of many chronic inflammatory diseases. TNFα activation of the nuclear factor κB (NFκB) signaling pathway particularly in macrophages has been implicated in many diseases. We demonstrate here that G-protein coupled receptor kinase-2 and 5 (GRK2 and 5) regulate TNFα-induced NFκB signaling in macrophages. RNAi knockdown of GRK2 or 5 in macrophages significantly inhibited TNFα-induced IκBα phosphorylation and degradation, NFκB activation, and expression of the NFκB-regulated gene macrophage inflammatory protein-1β. Consistent with these results, over-expression of catalytically active GRK2 or 5 enhanced TNFα-induced NFκB activity. In addition, we show that GRK2 and 5 interact with IκBα in intact cells as well as in vitro via the N-terminal domain of IκBα and that IκBα is a substrate for GRK2 and 5 in vitro. Taken together, these results demonstrate that GRK2 and 5 are important mediators of TNFα-induced NFκB activity and suggest that GRKs could be potential therapeutic targets for treating TNFα-mediated inflammatory diseases.
KIT Signaling Regulates MITF Expression through microRNAs in Mastocytosis and Mast Cell Proliferation (38.16)
Authors: Takemoto Clifford , Noel Pierre , Mendell Joshua , Wentzel Erik , Carter Melody , Metcalfe Dean D. , Lee Youl-Nam 
Published: January 2023

Abstract
Activating mutations in codon D816 of the tyrosine kinase receptor, c-KIT, are found in the majority of patients with systemic mastocytosis, an aggressive disease characterized by proliferation of abnormal mast cells. Constitutive activation of the KIT signaling pathway is critical to the transformed phenotype. The MITF transcription factor is essential for normal mast cell development. We found that MITF is highly expressed in malignant mast cells from bone marrow biopsies of patients with systemic mastocytosis with the D816V KIT mutation; however, it is not highly expressed in mast cells from bone marrow biopsies of patients with other hematologic conditions. We show that in both primary and transformed mast cell lines, KIT signaling markedly upregulates MITF protein expression. MITF mRNA levels do not change significantly with KIT signaling, suggesting post transcriptional regulation. We performed an array screen for KIT-regulated microRNAs in mast cells and found that miR-539 and miR-381 are downregulated by KIT signals; furthermore, they suppressed MITF expression through conserved sites in the MITF 3' UTR. We demonstrate the requirement of MITF for the transformed phenotype by inhibiting tumor growth in colony forming assays and in a murine xenograft model by knockdown of MITF with a shRNA-expressing lentivirus. This work demonstrates a novel regulatory pathway between two critical mast cell factors, KIT and MITF, mediated by microRNAs; dysregulation of this pathway may contribute to abnormal mast cell proliferation and malignant mast cell disease.
A fundamental role for Interleukin-21 in germinal center antibody responses (38.21)
Authors: Vogelzang Alexis , McGuire Helen , Sprent Jonathan , King Cecile 
Published: January 2023

Abstract
T cell help to B cells is a fundamental property of adaptive immunity, yet only recently have many of the cellular and molecular mechanisms of T cell help emerged. T follicular helper (TFH) cells are the CD4+ T helper cells that provide cognate help to B cells for high affinity antibody production in germinal centers (GC), and have recently been shown to give rise to lymphoid reservoirs of antigen specific memory T cells. TFH cells produce IL-21 and we have shown that IL-21 is necessary for GC formation. However the central role of IL-21 in GC formation reflected its effects on CD4+ T cells rather than on B cells. Furthermore, bone marrow chimeras confirm that IL-21R-deficient B cells are capable of mounting an effective IgG1 response to T dependent antigen. Using an antigen specific model where TCR transgenic CD4 T cells specific for the ovalbumin peptide were introduced into IL-21R-/- mice, we show that IL-21 responsiveness by T cells was important for optimal primary and secondary antibody responses. IL-21 also proved indispensable for affinity maturation of ova-specific antibodies in the GC. This study reveals a previously unappreciated role for TFH cells in the formation of the GC and isotype switching through a CD4+ T cell intrinsic requirement for responsiveness to IL-21.
Lymphotoxin-alpha beta heterotrimers are shed from the surface of activated human lymphocytes, circulate in serum, and are elevated in synovial fluid of rheumatoid arthritis patients (38.22)
Authors: Young Judy C , Yu Xin , Nguyen Allen , Kung Catherine , Wong Wai Lee , DeForge Laura , Grogan Jane 
Published: January 2023

Abstract
Lymphotoxin, a trimeric cytokine in the tumor necrosis family, is expressed by activated T, B, and NK cells and is involved in inflammatory response signaling and secondary lymphoid organ architecture. Using a specific assay for human LTαβ which detects both LTα1β2 and LTα2β1 trimers but not LTα3, we show here that LTαβ is detected in culture supernatants of 293 cells transfected with human LTα and LTβ constructs, and in supernatants of primary activated human T lymphocytes. The levels of soluble LTαβ detected in culture supernatants of human CD4+ Th1 polarized lymphocytes treated with an inhibitor of ADAM17 (TACE) protease, were reduced 3-6 fold compared to supernatants from untreated cells. This suggests that LTαβ cleavage is carried out by TACE. A 26 KDa MW form of LTβ was detected in activated human T cell culture supernatant by Western blot analysis using an anti-LTβ-specific antibody. Soluble LTαβ was detected in serum from normal human donors and rheumatoid arthritis (RA) patients at levels of approximately 100 pg/mL. Soluble LTαβ was also detected in synovial fluid from RA patients at significantly higher levels than in synovial fluid from osteoarthritis patients. Cleavage of LTαβ trimers from the cell membrane and release into circulation has not been previously described. Soluble LTαβ may play a role as an inflammatory mediator of autoimmune disease.
Research supported by Genentech Inc., S. San Francisco, CA
Evaluation of heterophilic antibody blocking agents in the development of sensitive immunoassays for IL-17AA, IL-17FF, and IL-17AF (38.24)
Authors: DeForge Laura E , Loyet Kelly M , Delarosa Donnie , Chinn Jason , Townsend Michael J , Wong Wai Lee T 
Published: January 2023

Abstract
The proinflammatory cytokines interleukin 17 (IL-17) A and F have been implicated in the pathophysiology of autoimmune diseases. In an effort to measure the concentrations of homodimeric IL-17AA, IL-17FF, and the recently identified IL-17AF heterodimer in patient samples, sensitive immunoassays for the three forms of IL-17 were developed in 384 well plates. Samples from autoimmune patients are particularly prone to false readings due to interference from heterophilic antibodies (HA, a term which will be used to encompass true HA as well as rheumatoid factor and human anti-animal antibodies). To evaluate HA interferences in these assays, sera were screened to identify samples that generated false positive signals. These samples were then treated with up to eight different commercial HA blockers. One blocking agent (IIR) was found to consistently eliminate or greatly reduce nonspecific signal and was chosen for use in the final assays. The limits of quantitation of the assays were 0.78 pg/ml, 6.14 pg/ml, and 5.12 pg/ml for IL-17AA, FF, and AF, respectively, with a minimum two-fold sample dilution. IL-17FF and IL-17AF levels were below detection limits for normal, Crohn's disease, and ulcerative colitis (UC) serum as well as osteoarthritis and rheumatoid arthritis (RA) synovial fluid. The greater sensitivity of the IL-17AA assay allowed detection of low levels of IL-17AA in &lt;50% of the UC and RA samples only.
Mercury chloride triggers human mast cells to release vascular endothelial growth factor (VEGF)-relevance to autism (38.26)
Authors: Kempuraj Duraisamy , Hogan Jennifer , Zhang Bodi , Kourelis Taxiarchis , Manola Akrivi , Theoharides Theoharis C 
Published: January 2023

Abstract
Autism Spectrum Disorders (ASD) are pervasive neurodevelopmental disorders of early childhood. There is no pathogenesis or curative therapy. Many ASD patients have "allergic" symptoms and food intolerance; moreover, the prevalence of ASD in patients with mastocytosis is 10-fold higher than the general population, suggesting mast cell involvement. A recent paper with data from the Vaccine Safety Datalink showed significantly increased rate ratios for ASD with mercury exposure from vaccines. We, therefore, investigated the effect of HgCl2 on VEGF secretion, intracellular calcium ion levels and viability of human cultured mast cells. HgCl2 induced statistically significant VEGF release from mast cells at 0.1-10 µM (n=3, p&lt;0.05), but without a dose-dependent relationship; viability was unaffected. To study the mechanism of action, we investigated intracellular calcium ions. HgCl2 increased intracellular calcium ion levels within 15 min. Mast cell stimulation by HgCl2, possibly in combination with other triggers such as viruses, stress hormones or toxins at a vulnerable young age to release VEGF, which is also vasodilatory could disrupt the gut-blood-brain barriers and permit brain inflammation contributing to ASD pathogenesis. This work was supported by Theta Biomedical Consulting and Development Co., Inc. (Brookline, MA).
Ikaros regulates IL-10 expression in CD4+ T cells (38.27)
Authors: Umetsu Sarah E , Winandy Susan 
Published: January 2023

Abstract
IL-10 is a regulatory cytokine critical for controlling inflammatory responses. However, little is known about the molecular mechanisms controlling Il10 gene expression, although GATA-3 is known to be involved in Il10 expression by Th2 cells. Here we show that Ikaros, a zinc finger DNA-binding protein, plays an important role in the regulation of IL-10 in T cells. Upon initial stimulation by T cell receptor, cells deficient in Ikaros (Ikaros null) express significantly lower levels of IL-10 compared to wild type T cells. In addition, under Th2 skewing conditions, which induce high levels of IL-10 production by wild type cells, Ikaros null T cells are unable to properly differentiate and produce only low levels of IL-10. The use of a retroviral vector system to express a dominant negative isoform of Ikaros in wild type Th2 cells represses IL-10 production but does not significantly alter the levels of transcription factors Gata3 and Tbet. Further, expression of Ikaros in Ikaros null T cells restores IL-10 expression. We found that Ikaros binds to conserved regulatory regions of the Il10 gene locus in Th2 cells, supporting a direct role for Ikaros in Il10 expression. In addition, we have evidence to suggest a coordinate role for Ikaros and Gata3 in the regulation of Il10. A direct interaction of these proteins is being examined. Thus, we suggest of novel role of Ikaros in regulating Il10 gene expression.
Analysis of human IL-4 and IL-13 dependent signaling through the IL-4/IL-13Ra1 receptor complex as measured by hTARC production in A-549 cells (38.29)
Authors: Miller Renee , Staples Megan , Febo Ruth , Burns John , Wu Chengbin 
Published: January 2023

Abstract
IL-4 and IL-13 are key TH2 cytokines that contribute to asthma pathogenesis by promoting airway hyper-responsiveness, cellular infiltration and mucus over-production. Both cytokines have similar overlapping functionalities likely the result of having a shared type II receptor complex composed of IL-4R and IL-13Rα1 chains. In the presence of hIL-13Rα1 alone, IL-13 binds to IL-13Rα1 with low affinity but upon IL-4R recruitment a high affinity heterodimeric complex is formed leading to cytokine dependent cellular events. IL-13 can also bind to hIL-13Rα2 with high affinity but the biological mechanism of cell signaling through this receptor remains unclear. Since the A-549 human lung epithelial cell line can respond to both IL-4 and IL-13 stimulation in the presence of recombinant human TNF-alpha (rhTNF-α),
IL-4/IL-13Rα1 expression in A-549 cells was investigated and the molecular basis for IL-13 induced human TARC (Thymus Activation Regulated Chemokine) production was determined. Furthermore, up-regulation of hIL-13Rα2 was observed on A-549 cells following rhTNF-α treatment, therefore, the contribution of IL-13Rα2 signaling as related to TARC production was additionally assessed. Our study indicates that A-549 cells can produce high levels of TARC in response to IL-4 and IL-13 suggesting this response is apparently a result of signaling through the IL-4R/IL-13Rα1 pathway and not the IL-13Rα2 pathway.
Interleukin-7 controls glucose uptake in T-Lymphocytes by regulating Hexokinase II gene expression (38.4)
Authors: Khaled Annette R , Chehtane Mounir 
Published: January 2023

Abstract
Peripheral lymphocytes homeostasis is controlled by cytokine signals of which interleukin-7 (IL-7) figures most prominently. Cells deprived of IL-7 exhibit low levels of Adenosine tri-phosphate (ATP). However, little is known of the mechanism by which IL-7 regulates metabolic activity in dependent cells. We reported that shortly after IL-7 withdrawal, T-lymphocytes decreased their glucose uptake. Here we show that T-cells deprived of IL-7 decreased the gene expression and protein levels of the glycolytic enzyme, Hexokinase II (HXKII), required for generating glucose-6-phosphate. Re-addition of IL-7 to cytokine deprived T-cells restored HXKII gene expression within two hours. This increase in HXKII expression correlated with an increase in glucose uptake. In contrast, gene expression of the glucose transporter GLUT-1 increased after sixty hours of IL-7 readdition to deprived cells, and the gene expression of phospho-fructokinase (PFK) in response to IL-7 showed minimal differences. Inhibition of HXKII activity, using a specific inhibitor, Bromo-pyruvate, decreased glucose uptake in presence, but not the absence, of IL-7 while overexpression of HXKII restored glucose uptake in the absence of IL-7. In summary, we conclude that IL-7 controls glucose uptake in T-lymphocytes by regulating the gene expression of HXKII, suggesting a novel mechanism by which IL-7 supports glucose metabolism in T-cells.
Poly (I:C) induces shedding of soluble TNFR1 from human airway epithelial cells via a TLR3-RIP1-ERK signaling pathway (38.9)
Authors: Yu Man , Levine Stewart  J. 
Published: January 2023

Abstract
Soluble tumor necrosis factor receptors (sTNFR) modulate TNF bioactivity by binding TNF. The type 1 TNF receptor (TNFR1), a key receptor for TNF signaling, is generated by either inducible cleavage and shedding of TNFR1 ectodomains or constitutive release of TNFR1 exosome-like vesicles. The goal of this study was to identify Toll-like receptor (TLR) signaling pathways that induce shedding of sTNFR1. We demonstrated that poly (I:C), a synthetic double-strand RNA (dsRNA) analog which signals via TLR3, induced sTNFR1 shedding from human airway epithelial (NCI-H292) cells in a dose-dependent manner over 2h, whereas ligands for TLR2, TLR4, TLR7, and NOD2 (nucleotide-binding oligomerization domain containing 2) did not. RNA interference experiments showed that poly (I:C)-induced sTNFR1 shedding was mediated via TLR3 and receptor interacting protein1 (RIP1). To further define the TLR3 signaling pathways that mediate sTNFR1 release, we showed that a specific, cell-permeable peptide inhibitor of ERK activation significantly attenuated sTNFR1 release by 55%. In contrast, neither dual oxidase 1 or 2, nor reactive oxygen species (ROS) mediated poly (I:C)-triggered sTNFR1 shedding. Furthermore, the pan-caspase inhibitor z-VAD-fmk attenuated sTNFR1 shedding by 28%, which suggests that caspase-dependent pathways also participate in this process. In conclusion, we show that poly (I:C), a viral dsRNA analog, induces sTNFR1 shedding from human airway epithelial cells primarily via a TLR3-RIP1-ERK signaling pathway, with a minor contribution from caspase-dependent pathways. This identifies a mechanism by which airway epithelial cells can modulate viral-induced airway inflammation.
This work was funded by the Division of Intramural Research in NHLBI/NIH.
A2A adenosine receptor mediates suppression in a mouse model of colitis (39.1)
Authors: Kurtz Courtney C , Naganuma Makoto , Alam Mohammad S , Feldman Sanford H , Linden Joel M , Ernst Peter B 
Published: January 2023

Abstract
Background: Adenosine accumulates in inflamed tissue and can suppress proliferation and cytokine production in helper T cells (Th). Th cells from mice lacking the A2A Adenosine Receptor (A2AAR-/-) do not function properly in the CD45RB transfer model of colitis. Methods: To examine the role of A2AAR in regulating colitis, we tested colonic myeloperoxidase (MPO) activity after adoptive transfer of Th cell susbsets from wildtype or A2AAR-/- mice. Th cells from mesenteric lymph nodes (MLN) of colitic mice were treated with an A2AAR agonist to assess effects on cytokine production. To investigate the role of A2AAR on myeloid cells in the CD45RB model of colitis, we adoptively transferred wildtype Th cells into RAG1/A2AAR double knockout (DKO) mice. Results: Colons of mice that received A2AAR-/- CD45RBhi cells with wildtype or A2AAR-/- CD45RBlo Treg had significantly greater MPO activity than mice that received wildtype CD45RBhi and CD45RBlo cells. Cytokine production by MLN Th cells from colitic mice was suppressed by an A2AAR agonist. Adoptive transfer of CD45RBhi Th cells into either RAG1-/- or DKO mice induced colitis but co-transfer of CD45RBlo Tregs attenuated colitis only in RAG1-/- mice. Conclusions: These data indicate that adenosine signaling through A2AAR plays an important suppressive role in colitis and that this is mediated through effects on both T cells and myeloid cells. CK supported by a CCFA Fellowship.