bluuebunny/crossref_metadata_2025_split huggingface dataset:
The variation of CD3+CD4-CD44HighCD62L- T cells in the development of asthma in mice (HYP7P.302)
Authors: Li Yanchun , Hua Shucheng , He Xiaoyan , Li Yun , Zhu Yaping 
Published: January 2023

Abstract
Objective CD44HighCD62L- is the marker for effector memory T cell. It is well-known that CD4+ T memory cells (Tms) play important role in asthma, but the role of CD8+ T memory cells (Tms) in asthma remains unknown. Here, we analyzed the variation of CD4-Tms in asthmatic mice so to have a general understanding of the role of CD8+Tms in asthma indirectly. Method To construct asthma mice model, BALB/c mice were sensitized and challenged with OVA for continuous 7 days. We detected the ratio of CD3+CD4-CD44HighCD62L- T cells in CD3+CD4-T cells from 1 day before challenge to 1 month after challenge by using a flow cytometry. Results In lungs, the ratio of CD4-Tms increased gradually after 1st challenge from 23.56% to 30.245% and reached the peak value with 61.97% at 2nd day after the last challenge. Interestingly and notably, it decreased rapidly to the level before challenge after that. This is quite different from CD4+Tms in which CD4+Tms maintained high level for much longer time. In spleens, the ratio of CD4-Tms maintained low level with about 20% during challenge. It increased to 35.86% after the last challenge and maintained at a stable level of 30% for 2 week then returned to the level before challenge. This variation is also different from CD4+Tms in spleens which is almost synchronize with CD4+Tms in lungs. Conclusion These results suggest that CD8+Tm may play a role in asthma with different mechanism from CD4+Tm, the detailed mechanisms warrant further exploration.
Signaling pathways in eosinophils that regulate allergic asthma and inflammation (HYP7P.304)
Authors: Stokes Kindra , LaMarche Nelson , August Avery 
Published: January 2023

Abstract
Allergic asthma and inflammation is a disease of the airways. The prevalence of allergic responses and asthma are increasing worldwide, but it is still unclear why some predisposed individuals develop disease. Eosinophils and T helper 2 cells (TH2) and eosinophils are critical cellular mediators in asthma. The signal transducer and activator of transcription (STAT)-6 is a crucial mediator of TH2 cytokine responses, acting downstream of interleukin -4 and -13 (IL-4 and IL-13). We have previously shown that eosinophil derived IL-13 plays an important role in the recruitment of T cells to the lung, and subsequent development of allergic asthma. However whether eosinophils respond to TH2 signals to control allergic airway inflammation is unclear. The objective of this study is to understand the role of STAT6 in eosinophil-dependent regulation of allergic asthma and inflammation. Using ΔdblGATA mice lacking eosinophils and transfer of STAT6-/- eosinophils, we found that compared to ΔdblGATA reconstituted with WT eosinophils, eosinophils lacking STAT6 were unable to rescue recruitment of CD4+ T cells and eosinophils into the lungs compared to WT eosinophils. These data indicates that STAT6 is important in the recruitment of both T cells and eosinophils to the lung during the development of allergic airway inflammation. Ultimately, deciphering the mechanisms of intracellular and cytokine signals will afford us improved insights on developing therapeutic strategies in asthma.
Fungal aeroallergen challenge results in increased granulocytic inflammation in Jh-/- mice in a murine model of allergic airway disease (HYP7P.280)
Authors: Ghosh Sumit , Asbach Scott , Hoselton Scott , Dorsam Glenn , Schuh Jane 
Published: January 2023

Abstract
B lymphocytes are well recognized in the development of IgE responses, which exacerbates asthma symptoms, but may also be important as a protective response against inhaled fungi. Further, these lymphocytes may play a direct role in the maintenance of airway wall fibrosis. Jh-/- mice have been used extensively to study the role of B lymphocytes in immune responses. These mice lack the Jh gene, which codes for the antibody heavy chain and is thought to be critical for B cell survival and function. In this study, we examined the role of B lymphocytes using a murine fungal aeroallergen model to mimic human fungal allergic airway disease. We compared disease progression in the BALB/c and Jh-/- mice and found no differences in airway hyperresponsiveness, collagen deposition, or overall pulmonary inflammation. However, the Jh-/- mice had a significantly higher number of neutrophils and eosinophils when compared to the BALB/c mice. IL-17A and IL-6 cytokines, which play a role in driving eosinophilia and neutrophilia, were also significantly higher in the Jh-/- mice after fungal challenge. We plan to characterize temporal activation and the relative contribution of these B lymphocytes in the maintenance of Tregs and Th17 lymphocytes within the pulmonary compartment. These studies will help expand our knowledge of how B lymphocytes regulate T lymphocyte responses in allergic asthma.
The Nrf2 activator, tBHQ, exacerbates immediate hypersensitivity response to food allergen (HYP7P.315)
Authors: Rockwell Cheryl , Gangur Venugopal , Pestka James , Para Radhakrishna , Turley Alexandra , Zagorski Joseph , Bursley Jenna , Dover Heather 
Published: January 2023

Abstract
Nuclear erythroid 2 related factor 2 (Nrf2) is a transcription factor activated by cell stress, including oxidative and electrophilic stimuli, resulting in the upregulation of cytoprotective genes. Nrf2 can be activated experimentally by a number of different pharmacological agents, including tert-butylhydroquinone (tBHQ), which is also used commercially as a food additive. We recently showed that activation of Nrf2 by tBHQ promotes Th2 differentiation and inhibits Th1 differentiation in isolated CD4+ T cells. Given that tBHQ is a food additive and promotes Th2 differentiation, the purpose of the present studies was to determine the effect of tBHQ in a mouse model of food allergy. Mice were administered a diet either with or without tBHQ (0.001%) for two weeks prior to sensitization with ovalbumin. Notably, this dose of tBHQ is lower than that normally present in standard rodent chow (0.0016%). The mice were sensitized weekly transdermally for 4 weeks. Sensitization to ovalbumin caused an increase in ovalbumin-specific IgE in plasma, which was greater in animals exposed to tBHQ. After sensitization, the mice were orally challenged with ovalbumin, causing mild/moderate immediate hypersensitivity, which was exacerbated by tBHQ. Overall, these studies suggest that low doses of the food additive, tBHQ, increase IgE response to food allergen and exacerbate clinical signs of immediate hypersensitivity. (This work was funded by NIH grant: ES018885.)
IgE and FcεRIα do not promote allergic airway disease in mice inoculated intratracheally with house dust mite extract (HYP7P.316)
Authors: McKnight Christopher , Perkins Charles , Finkelman Fred 
Published: January 2023

Abstract
The role of IgE in asthma pathogenesis is controversial. Early studies showed IgE contributed to allergic airway disease (AAD) when mice were sensitized passively by anti-ovalbumin (OVA) IgE or sensitized actively by OVA without adjuvant. Other groups found that IgE did not promote AAD that was provoked by fungal extracts or by OVA with adjuvant. We have used IgE-deficient mice and FcεRIα-deficient mice and their respective FVBN and BALB/c wild-type controls to study the IgE- and FcεRIα-dependence of AAD induced by a clinically relevant allergen and route of administration. Mice were inoculated intratracheally with house dust mite extract without adjuvant every 2 days for 7 doses. We measured airway hyperresponsiveness by both unrestrained plethysmography (Buxco) and forced oscillation (flexiVent), analyzed pulmonary inflammation by differential cell count on BALF, and sectioned lungs to quantitate goblet cell metaplasia by microscopic examination. We found no difference in any of these disease parameters between wild-type mice and gene-deficient mice, regardless of whether minimal, moderate, or more severe AAD was induced. These results complement the clinical observation that anti-IgE therapy fails to improve the underlying lung function of humans with long standing asthma, although this treatment does reduce acute exacerbations. We are currently studying the acute changes in lung function that occur upon inhalation of allergen in AAD and their dependence on the IgE axis.
Dermal exposure to the commonly used antimicrobial chemical triclosan promotes allergic responses in skin and lung (HYP7P.317)
Authors: Marshall Nikki , Long Carrie , Anderson Katie , Lukomska Ewa , Anderson Stacey 
Published: January 2023

Abstract
There has been a dramatic increase in the incidence of allergies and asthma in the U.S. thought to be influenced by environmental exposures. The antimicrobial chemical triclosan is found in countless products such as soap, lotion, toothpaste and kitchen utensils (0.1-1%). Triclosan is detected in the urine of the majority of Americans and recent epidemiological studies have found a positive association between urinary triclosan levels and allergic disease. Interestingly, exposure to triclosan, a non-sensitizer, enhanced allergic responses in a mouse ovalbumin (OVA) asthma model. To help elucidate the mechanisms behind the augmented allergic responses, triclosan (0-3%) was applied to the dorsal surface of the ears of BALB/c mice sensitized to OVA. Analysis of skin revealed infiltration of leukocytes and a dramatic increase in thymic stromal lymphopoietin (TSLP) expression even after a single dose. Analysis of cells in the draining LN of triclosan-exposed mice revealed a peak cell response at 7 days, including increased numbers and frequencies of GATA-3+ IL-4+ CD4 T cells, and CD11b+ CD80+ CD86+ MHC class II+ dendritic cells. Mice exposed to triclosan and challenged with OVA via airway aspiration also showed increased Th2 responses and OVA+ APC in their BAL fluid. These results suggest that triclosan augments allergic responses in skin and lung in part through mechanisms that involve TSLP and enhanced Th2 responses.
Reduced C epsilon transcription by stabilization of CD23 in B cells (HYP7P.320)
Authors: Fellmann Marc , Buschor Patrick , Röthlisberger Silvan , Stadler Beda , Vogel Monique 
Published: January 2023

Abstract
The low affinity receptor for IgE (CD23, FcepsilonRII) is part of the broad regulatory network controlling IgE synthesis. It occurs as membrane and as soluble receptor generated in vivo by specific proteolysis. Targeting membrane CD23 with either IgE or anti-CD23 antibodies is believed to inhibit IgE synthesis. However, the mechanism behind this negative feedback remains unclear. This mechanism was investigated by examining the effect of anti-CD23 molecules in form of designed-ankyrin-repeat-proteins (DARPins) specific for CD23. Previous experiments using anti-CD23 antibodies were difficult to interpret as their Fc portion also mediate feedback mechanisms. The DARPins were produced as monovalent and bivalent molecules that were able to compete with the binding of IgE to CD23. The effects of anti-CD23 DARPins were assessed in an in vitro system using anti-CD40/IL-4 stimulated B cells. Treatment of B cells with anti-CD23 DARPins resulted in stabilization of membrane CD23. The released amount of soluble CD23 was reduced while CD23 mRNA expression was unchanged suggesting that the anti-CD23 DARPins stabilize CD23 and prevent its proteolysis. IgE secretion was significantly reduced by anti-CD23 DARPins. In addition, analysis of the epsilon transcript confirmed the reduction of IgE synthesis whereas IgG secretion was not influenced by the anti-CD23 DARPins. Further experiments will be performed to assess signalling pathways triggered by CD23 cross-linking.
The association of complement protein C1q with apoptotic cells ingested by human dendritic cells modulates dendritic cell-mediated T cell activation (INC5P.332)
Authors: Clarke Elizabeth , Tenner Andrea 
Published: January 2023

Abstract
Humans deficient in complement protein C1q develop systemic lupus erythematosus (SLE) with nearly 100% penetrance, but the molecular mechanisms underlying this association have not yet been uncovered. Our previous studies have shown that human monocyte-derived immature dendritic cells (DC) ingesting C1q-bound autologous apoptotic cells (AC) (C1q-polarized DC) increase secretion of anti-inflammatory cytokines such as IL-10 and reduce pro-inflammatory cytokines such as TNFα. In this study, we demonstrate that C1q-polarized DC express significantly more surface PD-L2 and CD39 and significantly less CD86 relative to DC ingesting AC alone. To elucidate whether C1q-polarized DC functionally modulate the adaptive immune responses, human autologous T cells were incubated with DC or C1q-polarized DC and the proliferation and induction of CD4+ and CD8+ T cell subset was assessed. We discovered that relative to DC ingesting AC in the absence of C1q, C1q-polarized DC decrease autologous Th17 proliferation and induction and Th1 proliferation when the T cells were maintained in IL-7 prior to the MLR. These data imply that C1q-polarized DC can switch T cell function to reduce T effector cell activation and/or suppress the induction of autoimmunity, identifying new potential pathways for therapeutic intervention in SLE and other inflammatory diseases.
Deficiency in transient receptor potential melastatin 2 impairs alveolar macrophage activation and protects mice against endotoxin-induced lung inflammation and injury (INC5P.333)
Authors: Botta Davide , Mudunuru Uma , Southworth Kristen , Simpler Thomas , Bradley John , Randall Troy , Stevenson Christopher , Lund Frances 
Published: January 2023

Abstract
Cellular responses to reactive oxygen species (ROS) are induced by many receptors including the non-selective cation channel TRPM2 that promotes cation influx and membrane depolarization following ROS exposure, and potentiates inflammatory cytokine production and cell death. Conflicting reports on the harmful vs. protective roles of TRPM2 in ROS-mediated disease models led us to test whether TRPM2 is required for protection against lung endotoxin exposure, one of the most common etiologies of Acute Lung Injury (ALI). We found that Trpm2-/- mice exposed to intranasal lipopolysaccharide (LPS) were protected from lung inflammation and injury and that protection was due to loss of TRPM2 in radiation-resistant lung cells. Next, we examined the role of TRPM2 in radiation-resistant alveolar macrophages (AMs) as these cells are known contributors of ROS in ALI. Using congenic mice harboring the CD45.1/CD45.2 allelic variants we generated bone marrow chimeras in which radiation-resistant host and donor-derived AMs could be distinguished. We observed increased resistance to cell death in host Trpm2-/- AMs. Furthermore, LPS-activated Trpm2-/- AMs showed decreased expression of iNOS, inflammatory cytokines and proteases. Macrophage depletion experiments are underway to investigate the protective role of Trpm2-/- AMs in vivo. Our data support a harmful role of TRPM2 in ALI, and attribute the protective phenotype of Trpm2-/- mice to impaired macrophage-mediated lung inflammatory responses.
Changes in zebrafish phagocytic activity induced by acute ethanol exposure vary by time of day and leukocyte type (INC5P.327)
Authors: Macklis Paul , Curran Kaitlin , Newton-Acquah Antoinette , Lopez Juvenal , Hannum Lynn 
Published: January 2023

Abstract
Chronic alcohol abuse and acute alcohol consumption (binge drinking) have been associated with suppression of immune function. Reduced phagocytic activity in macrophages and altered levels of inflammatory mediators have been reported in mammalian studies of ethanol exposure. To explore whether the zebrafish model could be valuable in understanding the effects of ethanol on innate immune activity, we assessed the activity of zebrafish phagocytes after acute exposure to ethanol in tank water. Experiments were conducted at different times of day to also determine whether effects of ethanol vary in a diurnal pattern. Treating wild-type AB strain fish with ethanol in the morning reduced the percentage of total phagocytic cells that ingested bacteria. The opposite results were observed in the evening, when a higher proportion of cells participated in phagocytosis compared to controls. To distinguish between the activity of macrophages and neutrophils, mpx:GFP zebrafish were also used. Exposure to ethanol in tank water resulted in a reduction in the amount of bacteria engulfed by neutrophils, whereas changes in macrophage phagocytic activity were not as evident. These initial results indicate that ethanol affects subsets of phagocytic cells differently, and that the effects may be time-of-day dependent.
Activation-specific metabolic regulation of NK cell IFN-gamma production (INC6P.334)
Authors: Cooper Megan , Keppel Molly , Topcagic Nermina 
Published: January 2023

Abstract
Many factors that stimulate NK cell functional responses have been described; however metabolic fuels that drive NK cell function are unknown. Metabolism is significantly altered in disease states and metabolic regulation of T cell function is well-described. We hypothesize that NK cells require distinct metabolic fuels for survival, proliferation, and functional responses. NK cell metabolic phenotype was determined with an extracellular flux analyzer to determine basal oxidative phosphorylation (oxphos) and glycolysis of resting and activated NK cells. NK cells were then cultured under a variety of conditions using inhibitors and nutrient deprivation to determine cellular requirements for glucose, fatty acids, and oxphos for NK cell survival, proliferation, and function. NK cells primarily utilize oxphos at baseline and, in contrast to T cells, upon proliferation. However, there is significant plasticity in the metabolic requirements for NK cell IFN-gamma production. NK cell IFN-gamma production in response to cytokine stimulation is not dependent on either glycolysis or oxphos. By contrast, NK cell IFN-gamma production in response to receptor stimulation requires oxphos and is partially dependent on glycolysis. The metabolic requirements for NK cell IFN-gamma production are not altered by priming in vitro or in vivo, but do change with proliferation. These findings suggest that the metabolic microenvironment in vivo may significantly alter NK cell function.
Increased IRAK-M in peritoneal macrophages of NOD mice suggests in vivo exposure to bacterial endotoxins from the gut (INC6P.343)
Authors: Emani Rohini , Alam Catharina , Hanninen Arno 
Published: January 2023

Abstract
Macrophages play an important role in innate immune reactions, and peritoneal macrophages guard the sterility of this compartment mainly against microbial threat from the gut. Type-1 Diabetes (T1D) is an autoimmune disease in which gut microbiota and gut immune system appear to play a role. We have observed increased permeability and free radical production of gut epithelium in nonobese diabetic (NOD) mice. Impaired barrier function could lead to bacterial endotoxin leakage to peritoneal cavity. To seek for extraintestinal consequences of impaired barrier function, we characterized peritoneal lavage cells from young NOD mice after weaning. We detected a rapid increase in the number of macrophages in 1-2 weeks from weaning in NOD mice compared to C57BL/6 and BALB/c mice, and to NOD mice fed an antidiabetogenic diet (ProSobee), which improves gut barrier function. Upon LPS stimulation ex vivo, macrophages produced low amounts of TNF-α and showed reduced expression of CD40. Expression of TLR signalling pathway negative regulator, Interleukin-1 Associated Kinase -M (IRAK-M) was increased, indicating prior in vivo exposure to TLR-4 ligand(s). Upon further phenotyping, increased levels of ARG1 were observed suggesting in vivo differentiation towards M2-like phenotype. Bacterial endotoxins in peritoneal cavity may contribute to autoreactive T-cell activation by increased production of cytokines and activation of antigen-presenting cells in pancreatic lymph nodes.
NKp46 modulates NK immune synapse function (INC6P.346)
Authors: Hadad Uzi , Thauland Timothy , Butte Manish , Porgador Angel , Krams Sheri 
Published: January 2023

Abstract
Natural killer cells play an important role in the first-line defense against tumor and virus-infected cells. The activity of NK cells is tightly regulated by a repertoire of cell-surface expressed inhibitory and activating receptors. NKp46 is a major NK cell activating receptor that is involved in the elimination of viral infected cells. NKp46 also regulates interactions with other immune cells including T cells and dendritic cells. Recent studies revealed that complex integration of NK receptor signaling controls the cytoskeleton rearrangement and other immune synapse related events. However the distinct nature by which NKp46 participates in NK immunological synapse formation and function remains unknown. In this study we found that NKp46 forms microcluster structures at the immune synapse between NK cells and target cells. Labeling of F-actin showed that higher expression of NKp46 is correlated with increased accumulation of actin mesh at the immune synapse. Concordantly, knock-down of NKp46 in primary NK cells abridged recruitment of F-actin to the synapse. Live cell imaging experiments showed a linear correlation between NKp46 expression and lytic granule polarization to the immune synapse. Taken together, our data suggests that NKp46 signaling directly effects both the early formation and late function of the NK lytic immune synapse. Moreover, this is a continuous rather than threshold effect of activation vs non-activation.
Inhibition of PI3K contributes to nanoparticle-mediated exaggeration of endotoxin-induced leukocyte procoagulant activity (INC6P.340)
Authors: Ilinskaya Anna , Man Sonny , Patri Anil , Clogston Jeffrey , McNeil Scott , Dobrovolskaia Marina 
Published: January 2023

Abstract
Nanoparticles are increasingly used in biomedical applications due to their ability to change biodistribution, increase safety and improve efficacy of drugs. However, application of engineered nanomaterials in medicine is often halted by safety concerns. Endotoxin contamination is very common in nanomaterials and there is an increasing number of studies reporting that certain nanoparticles can exaggerate endotoxin-mediated inflammation. We studied the relationship between physicochemical properties of polyamidoamine (PAMAM) dendrimers and their ability to exaggerate LPS-induced procoagulant activity (PCA) of human leukocytes in vitro. The PCA is phospholipid-protein complex composed of tissue factor (TF) and phosphotidylserine (PS). We found that cationic but not anionic or neutral PAMAM dendrimers, increase LPS-induced leukocyte PCA. Cationic PAMAM dendrimers enhanced both LPS-induced TF expression and PS exposure on the cell surface. The mechanism of LPS-induced PCA exaggeration by PAMAM dendrimers is not mediated by the electrostatic interaction between cationic dendrimers and anionic LPS, calcium influx and secondary messengers released by dendrimer activated platelets. We hypothesize that dendrimers exaggerate LPS-mediated PCA through the inhibition of negative regulation of inflammation. Specifically, we demonstrate that cationic PAMAM dendrimers inhibit phosphoinositol-3-kinase (PI3K), a negative regulator of TF expression in response to endotoxin.