bluuebunny/crossref_metadata_2025_split huggingface dataset:
The microbiome regulates the eosinophil requirement for development of allergic lung inflammation (HYP2P.337)
Authors: Stokes Kindra , Wood Amie , August Avery 
Published: January 2023

Abstract
The diversity of gastrointestinal and respiratory microflora is essential in the shaping the host immune responses. In particularly, the propensity for Th2 allergic responses and asthma has recently been linked to the dysbiosis of the gut microbiome. Thus, the early establishment of particular microflora may potentially explain why some individuals are more predisposed to develop asthma and some do not. Eosinophils may also make differential contributions to allergic asthma in different individuals, and we have previously shown that development of allergic asthma is dependent on eosinophils. Here we report that C57Bl/6 ΔdblGATA eosinophil deficient mice housed under “less-restricted specific-pathogen-free” (LR-SPF) conditions are protected against allergic asthma compared to mice house under “restricted specific-pathogen-free” (R-SPF) conditions. 16s rRNA analysis of gut and lung microbiome reveal differences in lung microbiota diversity, suggesting a potential explanation for this phenomenon. Furthermore, consistent with the “microbiota hypothesis”, reducing the microbiota in ΔdblGATA mice by antibiotic treatment resulted in enhanced susceptibility to developing allergic airway inflammation comparable to WT mice. Thus, we conclude that shifts in microbiota can affect the requirement for eosinophil-mediated Th2 responses.
Alteration of human blood type 2 follicular helper T cells and regulatory B cells underlies comorbid association of allergic rhinitis with bronchial asthma (HYP2P.338)
Authors: Kamekura Ryuta , Shigehara Katsunori , Jitsukawa Sumito , Kawata Koji , Himi Tetsuo , Ichimiya Shingo 
Published: January 2023

Abstract
Allergic rhinitis (AR) is the most common allergic inflammatory disorder of the airway and its increasing worldwide prevalence has made it a global health problem. AR frequently precedes bronchial asthma (in 10 to 40% of cases), and the presence of AR is considered to be a risk factor for new onset of asthma. However, little is known about the mechanism by which AR advances to a state of AR with bronchial asthma (AR+Asthma). To determine the pathophysiologic features of AR and AR+Asthma, we examined subsets of follicular helper T (Tfh) cells and regulatory B (Breg) cells in peripheral blood from AR and AR+Asthma patients. The results demonstrated polarization of Tfh2 cells within Tfh cell subsets in both AR and AR+Asthma cases. Interestingly, Breg cells were decreased in AR cases and, more extensively, in AR+Asthma cases. Addtionally, we found significant correlations of fractional exhaled nitric oxide and blood eosinophil levels with the index %Tfh2 cells per %Breg cells. Our findings indicate that depletion of Breg cells under the condition of Tfh2 cell skewing is a putative exaggerating factor of AR to bronchial asthma.
Gene expression profile of allergen-experienced group 2 innate lymphoid cells (HYP2P.323)
Authors: Mathae Laura , Martinez-Gonzalez Itziar , Takei Fumio 
Published: January 2023

Abstract
We previously identified group 2 innate lymphoid cells (ILC2s) in mouse lungs. Upon allergen encounter, they produce type 2 cytokines and mediate T cell-independent allergic lung inflammation and also promote Th2 responses. We have recently found that allergen-treatments of mice result in significant increase in the number of ILC2s in the lung and lymph node even after the resolution of inflammation for several weeks. The allergen-experienced ILC2s are more responsive to unrelated allergens than naive ILC2s and rapidly induce allergic lung inflammation. To characterize the allergen-experienced ILC2s, we have performed microarray analysis of ILC2s purified from naïve mice and allergen-treated mice 4 days, 2 weeks, and 4 months after the treatment. The genes that are differentially expressed at different time points include those encoding chemokines, cytokines and their receptors, cell cycle and proliferation related proteins, adhesion molecules, and transcription factors. Gene Set Enrichment Analyses of the ILC2 populations revealed that allergen-experienced ILC2s are enriched for the gene signature of memory T cells. These results suggest that activated ILC2s undergo cell intrinsic changes and acquire memory-like features that are governed by the same set of genes as those responsible for memory T cell differentiation.
PARP inhibition by olaparib or gene knockout blocks asthma manifestation in mice by modulating CD4+ T cell function (HYP2P.324)
Authors: Ghonim Mohamed , Pyakurel Kusma , Ibba Salomi , Al-Khami Amir , Wang Jeffery , Rodriguez Paulo , Rady Hamada , El-Bahrawy Ali , Lammi Matthew , Ramsay Alistair , Mansy Moselhy , Ochoa Augusto , Naura Amarjit , Boulares Hamid 
Published: January 2023

Abstract
An important portion of asthmatics do not respond to current therapies. Thus, the need for new therapeutic drugs becomes urgent. We have demonstrated a critical role for PARP in experimental asthma. In the current study, we test the efficacy of Olaparib, a PARP inhibitor, recently introduced in clinical trials against cancer in blocking established asthma traits in animal models of the disease and determine the mechanism by which it achieves such protection. We show that administration of olaparib in mice post-challenge conferred a robust reduction in airway eosinophilia, mucus production and hyperresponsiveness even after repeated challenges with ovalbumin. The protective effects of olaparib were linked to a suppression of Th2 cytokines and ova-specific IgE with an increase in the Th1 cytokine IFN-g. These traits were associated with a decrease in splenic CD4+T cells and concomitant increase in T-regulatory cells. Adoptive transfer of Th2-skewed OT-II-WT CD4+T cells reversed both Th2 and Th1 cytokines and ova-specific IgE production in ovalbumin-challenged naïve PARP-1-/- mice suggesting a role for PARP-1 in CD4+T but not B cells. Our ex vivo studies show that olaparib treatment markedly reduced CD3/CD28-stimulated gata-3 and il4 expression in Th2-skewed CD4+T cells while causing an elevation in t-bet and ifn-g expression in Th1-skewed CD4+T cells. Our findings show the potential of PARP inhibition as a viable therapeutic strategy to be tested in human clinical trials
Sex-dependent differences in lung inflammation and M2-macrophage responses in asthma (HYP2P.328)
Authors: Keselman Aleksander , Heller Nicola 
Published: January 2023

Abstract
​Asthma is a chronic airway inflammatory disease that exhibits a sex difference. Alveolar macrophages acquire an M2-phenotype in asthma and in response to IL-4 in vitro. M2 macrophages correlate with poor lung function and pathology in asthmatics. We hypothesized that macrophages from female mice are intrinsically more responsive to IL-4 than male macrophages. Western blot demonstrated greater expression of the M2-marker, Ym-1 (~14 fold), in IL-4-activated bone marrow-derived macrophages (BMM) from female mice compared to male mice. We next assessed alveolar macrophages from male and female OVA-sensitized and -challenged mice for expression of M2-markers by FACS and western blot. We measured enhanced eosinophilia and increased Ym-1 expression in alveolar macrophages from female mice compared to male mice following OVA challenge. Estrogen enhances macrophage activation but the impact on lung macrophages during asthma is poorly understood. We observed a greater percentage of macrophages in the lungs of estrogen-implanted female mice that had undergone ovariectomy, compared to placebo-implanted females, following OVA challenge. These data suggest both intrinsic and estrogen-enhanced M2 macrophage polarization in females compared to males. Understanding intrinsic and estrogen-mediated sex differences in asthma is paramount to identify novel molecular targets and improve therapeutics in women with hard-to-treat asthma.
Repeated low dose injection of cobalt chromium microparticles in a mouse knee model induces robust inflammation with cartilage and bone loss (INC1P.345)
Authors: Palma Mark , Mishra Pankaj , Millman Ariel , Hallab Nadim , Beebe Kathleen , Benevenia Joseph , Gause William 
Published: January 2023

Abstract
Total joint arthroplasty is a well accepted orthopaedic procedure for the treatment of many pathological joint conditions, but the release of metallic wear debris such as cobalt chromium (CoCr) microparticles can lead to pain, osteolysis, and implant failure. Our studies of periprosthetic tissue samples from patients undergoing revision surgery have shown that these particles induce an M2 macrophage phenotype as evidenced through gene expression analysis and CD68 immunostaining. We used an intra-articular murine knee model to further examine the synovial tissue response to CoCr and have shown that a single high dose (10mg) inoculation of CoCr microparticles triggers a robust type 2 immune response characterized by the presence of alternatively activated macrophages, neutrophils, and eosinophils 2 days after injection. We further examined the long term effects of repeated alternate day low dose (0.5mg) injection of CoCr microparticles on synovial tissues over 14 days. Tissues exposed to CoCr microparticles were found to have increased clinical scores for inflammation with loss of cartilage and bone tissue at day 14 when compared to controls. Together these studies indicate that exposure of synovial tissues to CoCr induces an innate type 2 immune response with long term hard tissue damage. These studies may ultimately lead to the identification of targeted therapeutic options to locally treat implant associated pain and potentially increase implant longevity.
DOCK8 regulates protective immunity by controlling the function and survival of RORγt+ ILCs (INC1P.402)
Authors: Singh Akhilesh , Eken Ahmet , Fry Mallory , Bettelli Estelle , Oukka Mohamed 
Published: January 2023

Abstract
Retinoic-acid receptor-related orphan receptor-γt-positive (RORγt+) innate lymphoid cells (ILCs) produce interleukin (IL)-22 and IL-17, which are critical for protective immunity against enteric pathogens. The molecular mechanism underlying the development and survival of RORγt+ ILCs is not thoroughly understood. Here we show that Dedicator of cytokinesis 8 (DOCK8), a scaffolding protein involved in cytoskeletal rearrangement and cell migration, is essential for the protective immunity against Citrobacter rodentium. A comparative RNA sequencing-based analysis reveals an impaired induction of antimicrobial peptides in the colon of DOCK8-deficient mice, which correlates with high susceptibility to infection and a very low number of IL-22-producing RORγt+ ILCs in their GI tract. Furthermore, DOCK8-deficient RORγt+ ILCs are less responsive to IL-7 mediated signaling, more prone to apoptosis and produce less IL-22 due to a defect in IL-23-mediated STAT3 phosphorylation. Our studies reveal an unsuspected role of DOCK8 for the function, generation and survival of RORγt+ ILCs.
Respiratory syncytial virus disease is mediated by age-variable IL-33 and innate lymphoid cells (INC1P.347)
Authors: Saravia Jordy , You Dahui , Shrestha Bishwas , Lee Greg , Jaligama Sridhar , Rovnaghi Cynthia , Bagga Bindiya , DeVincenzo John , Cormier Stephania 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is the most common cause of infant hospitalizations and represents an enormous health burden worldwide. Severe RSV infections are a significant risk factor for childhood asthma, but the pathogenic mechanisms responsible for RSV induced airway disease remain elusive. We and others have shown that age at time of initial infection is a crucial determinant of RSV immunopathogenesis (i.e., Th2 inflammation, eosinophilia, airway hyperresponsiveness, mucus). Using an age-appropriate mouse model of RSV that phenotypically recapitulates severe Th2-biased RSV seen in human infants, we show that this age-related disparity in immunopathogenesis is related to enhanced IL-33 production and increased group 2 innate lymphoid cells (ILC2s) in lungs of neonatal mice compared to adult mice. In nasal aspirates from RSV-infected human infants, IL-33 was elevated in the youngest of patients which further indicates age-variable regulation in response to RSV infection. These data provide a previously unknown mechanism involving ILC2s and IL-33 in RSV mediated human asthma, and represent the first evidence of differential ILC2/IL-33 regulation with respect to very early age.
The protective role of γδ T cells in neonatal influenza (INC1P.348)
Authors: GUO XIZHI , Dash Pradyot , Calverley Matthew , Thomas Paul 
Published: January 2023

Abstract
Influenza virus infection is a continuing world-wide health threat, especially in infants and the elderly. Neonates are highly susceptible to influenza infection, which is often a cause of hospitalization. γδ T cells are the first T cells to develop during embryogenesis and dominate among the neonatal immune system components, as conventional αβ T cell responses are functionally less prominent. Hence, γδ T cells are poised to play a critical role in neonatal infections; however, the specific role of γδ T cells in neonatal influenza infection remains to be elucidated. Using a neonatal mouse model of intranasal influenza infection, we observed the accumulation of γδ T cells within two days after infection. By utilizing the Nur77-GFP reporter mice, we demonstrated that the activation of these γδ T cells was via the T cell receptor. IL-17-producing γδ T cells, rather than IFN-γ-producing cells, dominated in the primary response. A comparison between wild-type and TCRδ-deficient neonates identified a protective function of γδ T cells in neonates. Reduced weight loss and an increased survival rate were observed in the wild-type compared to TCRδ-deficient animals. In infected lungs, wild-type neonates also had more robust IFN-β production and increased inflammatory cytokine production. Our findings demonstrate a vital role for γδ T cells in mediating protection during neonatal influenza virus infection, which may provide a new therapeutic target for pediatric influenza treatment.
Microglial CX3CR1 is required for M2 polarization after intracerebral hemorrhage (INC1P.350)
Authors: Taylor Roslyn , Hammond Matthew , Ai Youxi , Sansing Lauren 
Published: January 2023

Abstract
Intracerebral hemorrhage (ICH) occurs when a blood vessel ruptures in the brain parenchyma, activating microglia. Once activated, microglia may polarize into an M1, pro-inflammatory phenotype, or an M2 phenotype associated with wound repair. Microglia express the chemokine receptor CX3CR1, which regulates microglial neurotoxicity. We have previously shown that mice with CX3CR1-null microglia have worse functional outcomes 14 days after ICH. We hypothesize that CX3CR1-null microglia fail to transition to an M2 phenotype and do not aid in tissue repair. C57BL/6 (WT) and CX3CR1GFP/GFP (CX3CR1-null) mice were irradiated and reconstituted with bone marrow from CD45.1 mice to make bone marrow chimeras. ICH was induced by injecting 20ul of WT whole blood into the right striatum. WT mice were sacrificed 1-14 days after ICH to examine changes in gene expression. Microglia were sorted and qRT-PCR was performed. Chimeras were sacrificed 14 days after ICH and perihematomal regions were harvested for intracellular staining (ICS). At 12 hours after ICH, microglia have high IL-6 gene expression (M1). However, M2 markers arginase-1, TGF-β and SOCS3 increase over time, suggesting microglia transition from M1 to M2 over the first 2 weeks after ICH. By ICS, CX3CR1-null microglia had significantly less CD206 (p&lt;0.01) and BDNF (p&lt;0.001) than WT microglia 14 days after ICH. Our data suggest that microglia transition from M1 to M2 after ICH and that CX3CR1 is required for M2 polarization.
Mesenchymal cell effect on the immune response in renovascular hypertension (INC1P.352)
Authors: Grande Joseph , Azimi Camillia , Hartono Stella 
Published: January 2023

Abstract
Atherosclerotic renal artery sclerosis (RAS) is a significant public health concern, affecting 7% of the population over 65 years old and conveying an increased risk for cardiovascular as well as renal disease. Recent reports have failed to show a benefit of renal artery stenting to treat RAS. We tested the hypothesis that mesenchymal stem cells (MSC) slow the progression of renal injury by reducing inflammation. MSCs were isolated and cultured from murine adipocyte tissue and injected via the carotid artery into the animals 4 weeks after inducing RAS, a time at which the stenotic kidney has undergone severe atrophy. Conjugated microsphere labeling and flow cytometry confirmed that the MSCs migrate to the stenotic kidney. Half of the treatment group demonstrated a 90% reduction in atrophy and significantly lower level of regulatory T cell (Treg) infiltration while the other half demonstrated no change when compared to vehicle treated RAS mice. Array analysis showed a significant down-regulation of NOS2 (M1 macrophages), RORc (Th 17 cells), and CCR4 (associated with M1 macrophages) within those mice with low renal atrophy. The results suggest that under certain conditions, MSCs have a possible effect on the inflammatory environment present in RAS mice. Future studies will determine whether intervention prior to development of severe atrophy will increase therapeutic efficacy of MSC.
Nucleotide binding characteristics of NOD2 proteins (INM1P.442)
Authors: Davis Beckley , Talento Suzanna , Tocker Aaron , Duncan Joseph 
Published: January 2023

Abstract
Nucleotide binding and oligomerization domain containing protein 2 (NOD2) is an intracellular protein that is involved in the recognition of bacterial cell wall derived muramyl-dipeptide. Mutations in the gene encoding NOD2 are associated with inherited inflammatory disorders including Crohn’s disease and Blau syndrome. NOD2 is a member of the Nucelotide-binding domain and leucine-rich repeat containing protein gene family (NLR). Nucleotide binding is thought to play a critical role in signaling by NLR family members. However, the molecular mechanisms underlying signal transduction by pathogenic NOD2 proteins remains unknown. Mutations in the nucleotide-binding domain of NOD2 have been shown to alter its signal transduction properties in response to muramyl-dipeptide in cellular assay. We demonstrate that nucleotide binding is necessary for wild type and pathogenic NOD2 protein activation of the NF-κB signaling pathway. Mutations of the Walker A motif inhibit binding to RIP2k in wild type NOD2, Crohn’s disease-associated NOD2 and Blau syndrome-associated NOD2. Loss of RIP2k binding correlates with decreased NF-κB-dependent luciferase activity. These data suggest that nucleotide binding is necessary for NOD2 function.
Neutrophil extracellular traps activated by damage associated molecular patterns exacerbates sterile inflammatory liver injury (INM1P.427)
Authors: Huang Hai , Tohme Samer , Al-khafaji Ahmed , Tsung Allan 
Published: January 2023

Abstract
The initiation of liver ischemia/reperfusion (I/R) injury results in the release of damage associated molecular patterns (DAMPs) such as HMGB1 and histones, which trigger innate immune and inflammatory cascade via Toll-like receptor 4 (TLR4) or TLR9. Infiltrated neutrophils contribute to the organ damage, innate immune and inflammatory responses after liver I/R. Formation of neutrophil extracellular trap (NET) has been recently found in response to various stimuli. However, the role of NETs during liver I/R remains unknown. We show that NETs form in the sinusoids of ischemic liver lobes in vivo, associated with increased serum level of myeloperoxidase (MPO)-DNA complexes and tissue level of citrullinated-histone H3 (NET markers) compared to control mice. Treatment with peptidyl-arginine-deiminase (PAD) 4 inhibitor or DNase I conferred significant protection after liver I/R evidenced by inhibition of NET formation, indicating the pathophysiological role of NETs in liver I/R injury. DAMPs, such as HMGB1 and histones stimulate NET formation through TLR4 and TLR9-MyD88 signaling pathways. After neutrophil depletion in mice, the adoptive transfer of TLR4 knockout (KO) or TLR9 KO neutrophils confers significant protection from liver I/R injury with significant decrease in NET formation. Conclusion: DAMPs released during liver I/R promotes NET formation through TLRs signaling pathway. NETs subsequently exacerbates organ damage and initiates inflammatory responses during liver I/R.
Rapid activation of lymph node lymphangiogenesis (IRC4P.458)
Authors: Ruddell Alanna , Farr Andrew , Jordan-Williams Kimberly 
Published: January 2023

Abstract
The lymphatic system delivers fluid and molecules from tissues to the lymph node (LN) to mediate immune system surveillance of self and foreign antigens. LN lymphatic endothelial cells (LECs) function in orchestration of the immune response, and in reinforcement of peripheral tolerance. Extensive LN lymphatic sinus growth (lymphangiogenesis) is involved in multiple diseases including cancer, arthritis, and inflammation. Lymphocytes and leukocytes regulate this lymphangiogenesis; however, the mechanism is not well defined. We identified a monoclonal antibody (10.1.1) that recognizes the LEC protein mCLCA1. Our studies identified mCLCA1 as an interacting partner for LFA-1 and Mac-1 that mediates lymphocyte and leukocyte adhesion. The 10.1.1 Ab blocks adhesion of lymphocytes to LECs in vitro to a greater extent than ICAM-1 neutralizing Ab, suggesting that while ICAM-1 interactions predominate in vascular endothelium, mCLCA1 interaction is more important for adhesion to LECs. In addition to mediating adhesion, the 10.1.1 Ab (but not control Ab) induces rapid LN lymphangiogenesis within 23 hours after injection into mice, which involves proliferation of LN LECs. This lymphangiogenesis is accompanied by LN remodeling involving multiple stromal cell types, resembling the LN response to inflammation. These findings suggest that 10.1.1 Ab mimics lymphocyte or leukocyte B2-integrin binding to LEC mCLCA1, to drive LN remodeling and lymphangiogenesis during immune responses.
Recovery of S100A8 in the absence of S100A9 exacerbates TNFα-mediated psoriatic-like arthritis (IRC4P.462)
Authors: Thurainayagam Sumita , Wixler Viktor , Roth Johannes , Vogl Thomas 
Published: January 2023

Abstract
Psoriasis is a chronic autoimmune disorder and frequently associated with arthritis (PsA). Alarmins S100A8 and S100A9, expressed in granulocytes, monocytes and activated keratinocytes, are highly up-regulated in psoriatic skin and synovium. Although S100a8 mRNA remains unchanged in S100A9-deficient (S100A9-/-) mice, S100A8 protein expression is abrogated, thus generating a functionally double knockout mouse model. In general, S100A9-/- mice show reduced inflammatory activities in several mouse models of infection and inflammation. Recently, we generated ihTNFtgxS100A9-/- mice by crossing doxycycline-inducible human TNFα-transgenic mice (ihTNFtg) with S100A9-/- mice. Unexpectedly, we observed an enhanced disease progression in these mice as evidenced by increased loss of body weight, paw swelling and cartilage destruction and decreased grip strength compared to ihTNFtg mice upon TNFα induction. Interestingly, the enhanced disease progression was associated with re-expression of S100A8 protein upon TNFα induction, observed in bone marrow, blood, skin and nail keratinocytes by IHC staining and optical imaging. We hypothesize that in the absence of its binding partner S100A9, constitutively active S100A8 homodimers aggravate TNFα-mediated psoriatic-like arthritis. Our data indicate a complex S100A8/S100A9 regulatory mechanism of an alarmin-driven inflammation.