bluuebunny/crossref_metadata_2025_split huggingface dataset:
Myosin-IIA regulates B cell leukemia migration and bone marrow engraftment and its inhibition reduces leukemia dissemination and prolongs survival (CAM4P.148)
Authors: Jacobelli Jordan , Wigton Eric , Thompson Scott 
Published: January 2023

Abstract
Leukemia relapses often affect peripheral organs such as the central nervous system (CNS), which acts as a sanctuary site for leukemia cells to escape anti-cancer treatments. Leukemia dissemination requires migration of cells from the blood circulation into target tissues by extravasation, similar to normal leukocyte migration. To extravasate, leukemia cells cross through vascular endothelial walls via a process called trans-endothelial migration (TEM), which requires cytoskeletal remodeling. However, the specific cytoskeletal effectors of leukemia extravasation are not fully known. We examined the role of Myosin-IIA (MyoIIA), a class II myosin motor protein, in leukemia cell extravasation and if its inhibition would hinder leukemia dissemination into the CNS. We inhibited MyoIIA expression using shRNA in a mouse model of Bcr-Abl driven B cell acute lymphoblastic leukemia (B-ALL). MyoIIA depletion did not affect apoptosis or the growth rate of B-ALL cells. Instead, MyoIIA was required for B-ALL cell chemotaxis towards CXCL12 and for completion of TEM. Inhibition of MyoIIA also reduced the ability of B-ALL cells to efficiently engraft and disseminate in vivo, including a highly significant reduction in CNS infiltration. Finally, MyoIIA inhibition prolonged survival in an in vivo leukemia transfer model. Our data implicate MyoIIA as a key regulator of leukemia cell migration making it a promising target to inhibit leukemia dissemination in vivo and reduce relapse frequency.
Tracking of CD8+ effector T cells in intact lung: analysis of environmental guidance (CAM4P.151)
Authors: Mrass Paulus , Ellenwood Sean , Moses Melanie , Cannon Judy 
Published: January 2023

Abstract
Effector T lymphocytes patrolling peripheral tissues protect organisms against infection and cancer. Recent two-photon imaging studies have revealed that T cells move with high speed through the interstitium of inflammatory tissues. We are interested in assessing migration strategies of tissue-infiltrating T cells and determining how T cell motility affects the efficiency of target search. We perform two-photon imaging using a murine endotoxin lung model to define the rules that govern motility of CD8+ effector T cells. We track individual T cells moving in intact lung for up to three hours but we do not find any evidence for long-range guidance cues for T cells, consistent with observations in other environments. Nevertheless, computational analysis of trajectories with an automated image-analysis algorithm reveals a significant alignment of T cells moving in conjunction with lung-associated vessels. Our results suggest that the architecture of the lung actively guides the migration of effector T cells and may act to limit their movement within tissues. Long term tracking of T cells moving in live tissue can provide a better understanding of the underlying mechanisms driving the search-efficiency of T cells during the effector phase of an immune response.
The TNF-family cytokine receptor death-receptor 3 (TNFRSF25) is required for nephritis but not systemic autoimmunity in the lpr murine model of lupus (CCR5P.209)
Authors: Hayes Erika , Villarreal Sarah , Ferdinand John , Engel Odile , Meylan Francoise , Siegel Richard 
Published: January 2023

Abstract
DR3 (Tnfrsf25) is a death domain-containing tumor necrosis factor receptor found primarily on activated T cells and innate lymphocytes. Interaction with its ligand, the TNF-family cytokine TL1A, plays a critical role in diverse autoimmune disease models in facilitating T cell accumulation at the site of inflammation while not affecting the systemic response. To determine whether DR3 could play a role in systemic autoimmunity, we studied the role of DR3 in the MRL/lpr mouse model of lupus. Lpr mice have a mutation in the TNF family receptor Fas (CD95, Tnfrsf6) that impairs its expression, causing systemic autoimmune disease that mimics systemic lupus erythematosus (SLE) observed in humans. DR3 deficient lpr mice developed identical degrees of lymphadenopathy, splenomegaly and double-negative T cells compared to control littermates. Expansion of effector T and B cell subsets were also not dependent on DR3. However, DR3 deficiency significantly protected MRL/lpr mice from IgG and C3 glomerular immune complex deposition and DR3 deficient kidneys had decreased medullary T cell accumulation, decreased periglomerular and medullary macrophage infiltrates and reduced proteinuria. These findings show that DR3 specifically promotes renal pathology in the MRL/lpr mouse model of lupus nephritis without affecting systemic autoimmunity or other features of Fas deficiency. Blockade of TL1A-DR3 interactions may be a novel strategy to counter nephritis and other organ damage in human SLE.
Study of the biological activities of a novel monoclonal anti-mouse CX3CR1 antibody, clone SA011F11. (CCR5P.213)
Authors: Lin Leo , Soto Hortensia , Oida Takatoku , Yang Xifeng , Moyron-Quiroz Juan 
Published: January 2023

Abstract
CX3C chemokine receptor 1 (CX3CR1) is a 7 transmembrane G-protein coupled receptor involved in the recruitment of cells to inflamed sites, cell adhesion, and cell extravasation; its ligand is CX3CL1. It has been reported that CX3CR1 is expressed on different cell types such as resident and alternatively activated macrophages, microglia, subsets of monocytes, DCs, NK cells, memory T cells, B cells, mast cells, astrocytes, and some tumor cells. Here we describe a novel anti-mouse CX3CR1 antibody, clone SA011F11, which is suitable for flow cytometry applications. In addition, to test the biological activity of this antibody in vitro, a chemotaxis assay was performed and the migration of CX3CR1-transfected BA/F3 cells was blocked by this clone with an ED50 of 0.11μg/ml. To test the effect of this antibody in vivo, a group of mice were injected i.p. with SA011F11 for 4 days and the changes in cell populations of different organs (spleen, peritoneal cavity, lungs, brain, and bone marrow) were studied. The administration of clone SA011F11 resulted in the reduction of CX3CR1+ cells in most of the analyzed organs and, although the degree of reduction varied, was more pronounced in the spleen and lungs while showing little effect in the brain. These findings demonstrate that clone SA011F11 has a biological activity in vivo and in vitro, and suggest that it is a powerful tool to study the biology of the CX3CL1/CX3CR1 axis.
Role of LFA-1 for protective immunity elicited by heterologous prime-boost vaccination against intracellular parasitic infection (CCR5P.206)
Authors: Vasconcelos Jose Ronnie , ferreira Camila Pontes , Rodrigues Mauricio 
Published: January 2023

Abstract
Prime-boost vaccine with DNA plasmid followed by adenovirus 5 is a powerful strategy to elicit specific CD8+T cells which play a key role in protective immunity (PI) against intracellular pathogens, such as Trypanosoma cruzi, the causative agent of Chagas' disease. Integrins, CAM and chemokine receptors may also play a role in this process. We demonstrated that treatment with anti-LFA-1 antibody, in immunized/infected mice, reverted the PI after experimental systemic infection. The aim of this work was to study the role of LFA-1 and VLA-4 integrins, the ICAM-1 and CCR5 after prime-boost vaccine. In order to study the role of LFA-1 and VLA-4 integrins, A/Sn mice were immunized with prime-boost vaccine. After, they were infected and treated with anti-LFA-1 and/or anti-VLA blocking antibodies. The immunized/infected mice treated with anti-LFA-1 displayed high parasitemia and all did not survive more than 30 days after challenge. Interestingly, treatment with anti-VLA-4 did not alter the protective response and all mice remained alive. In addition, LFA-1 blockage neither diminished the frequency nor compromised capacity of specific CD8+T cells in producing IFN-γ and/or TNF-α. Also, deficient ICAM-1 or CCR5 mice immunized and/or infected did not become susceptible to challenge. We concluded that LFA-1 play a critical role in the protective immune response generated by the vaccination, and by blocking this molecule, the CD8+ T cell migration capacity is likely impaired.
Soluble IL-15 complexes are generated after CD40 stimulation and VSV infection independent of type I IFN signaling. (CCR6P.218)
Authors: Anthony Scott , Schluns Kimberly 
Published: January 2023

Abstract
Interleukin (IL)-15 is a cytokine that promotes the development and homeostasis of a group of lymphocytes but is also significantly up-regulated in response to immune stimulation and during inflammation. Previous studies show that TLR stimulation and total-body irradiation (TBI) induce soluble IL-15Rα/IL-15 complexes (sIL-15), which are capable of mediating agonistic IL-15 effects in a paracrine manner. The effects of elevated IL-15 are well characterized, yet little is known about the signals inducing these complexes. We report that treatment with αCD40 Ab but not α41BB Ab led to systemic increases in sIL-15 complexes in vivo. BATF3 KO mice, which are deficient in CD8+ DCs, were impaired in the generation of sIL-15 complexes in response to αCD40 treatment, indicating an important role for CD8+ DCs in the generation of sIL-15 complexes in vivo. Interestingly, although CD40 stimulation or VSV infection enhanced systemic IFN-α protein expression, Type I IFN signaling was not required for the induction of sIL-15 complexes during either event. This is in contrast to the requirement for IFNAR signaling in generating sIL-15 complexes after TBI or Poly I:C. CD40 signaling during VSV infection was also not required for sIL-15 complex generation, as blockade with αCD40L exhibited no effect. Overall, this study indicates that sIL-15 complexes are induced in vivo by both type I IFN dependent and independent pathways.
Strong TCR signaling prolongs lineage uncertainty during MHC-I specific positive selection (HEM8P.241)
Authors: Kimura Motoko , Nakayama Toshinori , Singer Alfred 
Published: January 2023

Abstract
CD4/CD8 lineage choice in the thymus is primarily determined by the duration of TCR signaling during positive selection. Positive selection signaling by MHC-I ligands is of relatively short duration because CD8 coreceptor expression is terminated. However, the duration of TCR signaling during MHC-I specific positive selection should, in theory, also be affected by TCR affinity as well as coreceptor signal strength, but how TCR affinity and coreceptor signal strength affect MHC-I specific positive selection is not really known. To address this point, we used RAG-deficient mice expressing a panel of MHC-I restricted TCR transgenes with TCRs of different affinities for in vivo self ligands, and used genetically engineered mouse models that modulate the intensity of TCR signaling. Our data reveal that TCR signal intensity dictates how long MHC-I signaled thymocytes remain lineage-uncommitted before they express Runx3 and undergo CD8 lineage commitment. Furthermore, our data reveal that lineage errors by very high affinity TCR undergoing MHC-I specific positive selection do not normally occur because CD8 coreceptors have evolved to provide only weak Lck/coreceptor signals. We conclude that TCR signal intensity dramatically affects the timing of lineage fate during MHC-I specific positive selection and that error-free CD8 lineage choice is dependent on weak signaling by the CD8 coreceptor.
The hyperinflammatory reactivity of post-asthma alveolar macrophages contributes to RSV-induced asthma exacerbations (HYP7P.269)
Authors: Grooten Johan , Schepens Bert , Smet Muriel , Saelens Xavier , Naessens Thomas 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) causes serious morbidity as a major etiological agent of allergic asthma exacerbations. We now report that mice that suffered a preceding allergic bronchial inflammation were more prone to develop severe inflammatory responses and AHR after RSV-infection. Post-asthma alveolar macrophages (AM), residing in the alveolar lumen after clearance of eosinophilia, were found to largely contribute to this RSV-induced inflammation exacerbation. This AM population exhibited increased proinflammatory cytokine secretion in response to RSV-challenge but failed to contribute to viral clearance. A selectively increased proinflammatory NF-κB p65 signaling along with unaltered antiviral type I IFN signaling and loss of viral phagocytic clearance appear to be at the basis of the post-asthma AM mediated inflammation exacerbation. Phenotypic analysis revealed that the majority of post-asthma AM consisted of immature SiglecFlowCD11blow/int cells. Full maturation of the immature SiglecFlowCD11blow/int post-asthma AM by intratracheal administration of GM-CSF was associated with a decrease in RSV-induced exacerbation parameters, including pulmonary inflammation and AHR. These results point to a cascade of events in which a prior allergic bronchial inflammation causes a maturation defect in post-asthma AM resulting in a hyperinflammatory reactivity of the cells to RSV-infection, which in turn contributes to the onset of RSV-induced asthma exacerbation.
Development of a novel transcutaneous delivery peptide and its application in skin inflammation (HYP7P.273)
Authors: Kim WonJu , Choi Je-Min 
Published: January 2023

Abstract
Transcutaneous delivery of therapeutic drugs has many advantages over the systemic administration for inflammatory skin diseases such as atopic dermatitis or psoriasis. However, the greatest challenge for transcutaneous delivery is methodology due to the tight junction of skin tissue. Here, we screened and identified transdermal delivery peptide (TDP) from human proteins which would be non-invasive and simple method to deliver various cargos such as chemicals, peptides, proteins, etc. TDP-EGFP recombinant protein was purified and exhibited significantly higher intracellular transduction efficacy compared with TAT-EGFP in Jurkat, HaCaT, NIH3T3 cell lines and also in primary cells including splenocytes and keratinocytes. TDP-TAMRA peptides were successfully delivered into the epidermis and dermis of the mouse ear and back skin. We also used TDP-VIVIT peptide to inhibit T cell activation and contact dermatitis in vivo. TDP-VIVIT significantly inhibited IL-2 secretion with reduction of CD69 and CD44 expression level of activated CD4 and CD8 T cells. Our results suggest TDP-VIVIT peptide could be a potential transdermal applicable therapeutic drug for inflammatory skin diseases.
Overexpression of IL-18 promotes iNKT cell mediated eosinophilic esophageal inflammation in experimental murine model of eosinophilic esophagitis (HYP7P.257)
Authors: Upparahalli Venkateshaiah Sathisha , Jalahalli Mariswamy Siddesha , Shukla Anshi , Mishra Anil 
Published: January 2023

Abstract
Elevated level of IL-18 is reported in a number of allergic diseases and we earlier reported induced blood IL-18 and esophageal IL-18Rα mRNA in human EoE. Therefore, the current study focuses on the hypothesis that allergen-induced IL-18 and its receptor (IL-18Rα) positive cells may have an important role in EoE pathogenesis. Accordingly, we examined the blood IL-18 and esophageal IL-18Rα mRNA levels in aeroallergen and food allergen induced experimental mouse model of EoE. Herein, we demonstrate that blood IL-18 protein and esophageal IL-18Rα mRNA are induced in the mouse model of EoE and the source of induced esophageal IL-18Rα transcript are iNKT cells. We report that intranasal delivery of rIL-18 induces both mast cells and eosinophilic inflammation in the esophagus. Furthermore, to establish the significance of IL-18 in EoE pathogenesis, we next examined DOX-inducible rtTA-CC10-IL-18 bitransgenic mice that have significantly induced IL-18 protein in the esophagus. Our analysis indicated that DOX exposed IL-18 tg mice develop most of the characteristics of EoE, such as esophageal intraepithelial eosinophilia, increased mast cells and esophageal remodeling including fibrosis. Mechanistically, we show that IL-5 gene-deficient mice and iNKT cell-deficient (CD1d null) mice do not induce EoE in response to IL-18 intranasal challenge. Taken together, we provide evidences that allergen induced IL-18 has a significant role in promoting IL-5 and iNKT dependent EoE pathogenesis.
Sphingosine 1-phosphate receptor 3 is highly upregulated on human neutrophils upon migration to the airways (INC6P.315)
Authors: Ingersoll Sarah , Forrest Osric , Brown Milton , Tirouvanziam Rabindra 
Published: January 2023

Abstract
Background and hypothesis: Migration of polymorphonuclear neutrophils (PMNs) from blood into the airway lumen is accompanied by changes in surface receptors and responsiveness to environmental cues, such as sphingolipids. In particular, signaling via extracellular ceramide / sphingosine 1-phosphate (S1P) was suggested to contribute to human airway disease, such as cystic fibrosis (CF). We hypothesized that airway PMNs would alter expression of the S1P receptor, S1PR3, which was recently implicated in lung inflammation and fibrosis. Methods: We collected blood and sputum from healthy control (HC) and CF subjects and analyzed S1PR3 expression in live PMNs gated by flow cytometry. In addition, we used a novel in vitro model to study the effect of transepithelial migration of blood PMNs on S1PR3 expression. Results: We observed a 10-fold upregulation of surface S1PR3 expression on HC and CF airway PMNs, in vivo, compared to their blood counterparts. This major increase was recapitulated in vitro, when naive blood PMNs were induced to transmigrate through an airway epithelial layer toward apical CF airway fluid. S1PR3 expression increased over time, by 10-fold after 10 hours, and up to 20-fold at 18 hours. Conclusions: Modulation of S1PR3 expression follows PMN migration to both HC and CF airways. Further studies will determine the importance of this receptor in mediating responses to ceramide/S1P signaling in these cells, including PMN migration, apoptosis and activation.
Aging-associated chronic mast cell activation by aged mesenteric lymphatic vessels (INC6P.324)
Authors: Gashev Anatoliy , Nizamutdinova Irina , Chatterjee Victor , Cromer Walter , Tobin Richard , Zawieja David , Newell Rogers Karen 
Published: January 2023

Abstract
Here we present novel data related to interactions between lymphatic vessels, mast cells (MCs), eosinophils, and immune cells. MCs are known to inhabit in greater density the tissue near mesenteric lymphatic vessels (MLVs). Acute activation induces MC degranulation with subsequent recruitment of eosinophils and of MHC class II+ cells towards MLVs. Aging permanently influences mesenteric MCs by inducing chronic basal activation, thereby influencing contractility of aged MLVs. At the same time, the number of intact MCs available to react acutely to inflammatory stimuli decreases with age. The diminished number and activity of MCs in aged tissues also diminishes traffic and activation of eosinophils that are attracted to aged MLVs. Concurrently, MCs can function as helper cells for B cells that can produce IgE. IgE has a relatively short half-life in the absence of chronic MC activation. We propose that a key component of aging-associated alterations of “mesenteric lymphatic system-mast cells/eosinophils-inflammatory cells” system is the increased basal activation of resident MCs in aged mesentery that promote chronic inflammation with lesser potential to recruit enough anti-inflammatory MHC class II+ cells and eosinophils towards MLVs, resulting in the inability to prevent a smooth transition from acute inflammation to specific immune cell activation.
Development of uptake capacity and bactericidal activity in myeloid cells during GM-CSF-driven differentiation. (INC6P.326)
Authors: Olatunde Adesola , Landuyt Ashley , Abell Laura , Schwartz Elizabeth 
Published: January 2023

Abstract
The antigen presenting function of dendritic cells (DC) is well defined, yet their ability to handle bacterial infection is poorly understood. We have previously shown that GM-CSF-derived DC can confine Listeria monocytogenes (Lm) to the phagosome, preventing its invasion of the cytosol, growth, and spread. However, it is not clear at what stage of development DC acquire the capacity to retain Lm in the phagosome. To measure this activity at different stages of development, GM-CSF driven bone marrow cells were sorted based on expression of Ly6C, CD115, and CD11c. This strategy enables isolation of cells representing five distinct myeloid cell types: CMP, GMP, monocytes, inflammatory dendritic precursors (IDP), and inflammatory DC (IDC). To measure bacterial invasion of the cytosol, we used an Lm strain that expresses GFP upon cytoplasmic entry, in combination with fluorescent bioparticles to measure bulk uptake. The CMP demonstrated minimal uptake capacity and little bacterial cytoplasmic entry. The GMP population showed greatly increased uptake capacity, but also high susceptibility to bacterial cytoplasmic entry. In the monocyte population, the uptake capacity reached its peak, and these cells gained the capacity to confine the bacteria to the phagosome. Finally, the uptake capacity decreased modestly in the IDP and IDC populations, but these cells also blocked cytoplasmic invasion of the bacteria. The mechanistic basis of phagosomal retention was further examined.
Association of tissue transglutaminase and NLRP3 inflammasome in liver inflammation after Schistosoma japonicum infection (INM6P.343)
Authors: Li Zi , Ji Xiaofang 
Published: January 2023

Abstract
Liver diseases caused by Schistosoma japonicum(Sj) infection is one of the most serious pathological changes. Liver inflammation underlies infection-induced fibrosis, cirrhosis and liver cancer but the processes that promote hepatic inflammation by Sj are not well defined. We indicate that interleukin-1β (IL-1β) confers liver inflammation through Sj-induced NLRP3 inflammasome signaling, and tissue transglutaminase (tTG) is the upstream molecule of this signaling. Five weeks after Sj infection, elevated IL-1β levels in mice liver and also in serum compared to non-infection controls coincide with the expression level and activation of caspase-1 and tTG in mice liver. Immunohistochemical analysis of normal control and Sj-infected mice liver sections revealed that infiltrating inflammatory cells around the granuloma and Hepatic stellate cells (HSCs) among hepatic cells, are the primary cellular source of hepatic IL-1β and caspase-1 p10 during Sj infection. Sj infection triggers MyD88-mediated TLR4 signaling to induce IL-1β mRNA expression. Administration of cystamine, an inhibitor of tTG, abrogates the increase of IL-1β and caspase-1 p10 in infected mice. This result establishes a novel link between tTG and NLRP3 inflammasome under Sj infection. The important function of NLRP3 inflammasome activation and tTG in Sj infection makes them potential drug targets in preventing Sj-induced liver diseases.