bluuebunny/crossref_metadata_2025_split huggingface dataset:
Aryl hydrocarbon receptor modulates the development and activity of innate-like B cells (HEM1P.226)
Authors: Yeramilli Venkata , Tanner Scott , Martin Colin , Kearney John 
Published: January 2023

Abstract
The Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that recognizes polyaromatic hydrocarbons. While the contributions of AhR ligands to the differentiation and functions of Th17/Treg cells are relatively well understood, little is known about AhR in B cells. Microarray and quantitative PCR analysis of B cell subsets revealed that B1 and marginal zone (MZ) B cells expressed higher levels of AhR compared to follicular (FO) B cells. We found ~ two-fold more B1 B cells, but fewer MZ B cells in AhR-/- mice compared to AhR+/+ mice. Following immunization with Streptococcus pneumonia, AhR-/- mice had significantly higher titers of anti-phosphorylcholine IgM antibodies. In contrast, after immunization with a model T cell-dependent antigen, Trinitrophenyl Keyhole Limpet Hemocyanin (TNP-KLH), AhR-/- mice had anti-TNP titers that were comparable to those in AhR+/+ mice. Further, we found significantly fewer B cells in C57BL/6 mice raised on AhR ligand free diet, and an increase in titers of IgM and IgG3 in mice following a switch to an AhR ligand free diet when compared to littermates raised and maintained on conventional diet. Taken together, these findings suggests that AhR plays a role in the development of B1 and MZ B cells and regulates T cell-independent but not T cell-dependent humoral response. Further studies using mice with a B cell specific deletion of AhR are underway to elucidate the mechanism by which AhR ligands influence humoral immunity.
Human umbilical cord blood γδ T cells expanded ex vivo with zoledronic acid are distinct from human peripheral blood γδ T cells (HEM2P.244)
Authors: Tomchuck Suzanne , Evans Scarlett , Perko Ross , He Jin , McKenna Amy , Dash Pradyot , Thomas Paul , Dallas Mari 
Published: January 2023

Abstract
Ex vivo expansion of human peripheral blood (PB) γδ T cells using aminobisphosphonates has been extensively studied. However, as human umbilical cord blood (CB) γδ T cells are not as well described, we conducted a detailed comparison of proliferation, phenotype, TCR repertoire and function of CB and PB γδ T cells cultured with zoledronic acid. The percent of γδ T cells within the lymphocyte subset significantly increased in both samples after culture, but CB γδ T cells proliferated more than PB. Phenotypically, Vγ9Vδ2 was a common clone in fresh PB γδ T cells, which predominated after culture, yet the number of CB Vγ9Vδ2 cells remained low. The memory subsets of fresh γδ T cells were similar, but after culture, PB γδ T cells became traditional memory cells while CB γδ T cells were intermediate CD45RA+CD45RO+ memory cells. Memory status corresponded with TCRγδ variant; more Vγ9 and Vδ2 cells were memory while Vδ1 cells were naïve. Functionally, PB and CB γδ T cells had distinct cytokine secretion profiles. PB γδ T cells secreted more IFN-γ and TNF-α after culture, while CB γδ T cells secreted more IL-10. Using a single cell multiplex PCR assay, we found the TCRγδ repertoire of fresh cells to be diverse in both samples, yet CB γδ T cells maintained diversity after culture. Our results suggest that zoledronic acid may not stimulate γδ T cells in CB the same as PB and analysis of the TCRγδ variants may give insight into the difference in γδ T cell function of these samples.
Regulation of c-Myc/survivin from notch signaling modulates T cell differentiation (HEM2P.250)
Authors: Song Jianxun , Fino Krisitn , Haque Mohammad 
Published: January 2023

Abstract
Notch signaling is essential for T cell lineage commitment and is further required during early phases of thymocyte differentiation. Active Notch signaling during these early stages of T cell development inhibits other lineage potentials, such as B cell and myeloid cell (e.g., dendritic cell) potential. However, the intracellular signaling pathways by which Notch signaling regulates T cell differentiation remain unknown. Here we show that the transcriptional factor c-Myc is controlled by Notch signaling which regulates T cell differentiation. In a well-established in vitro differentiation of T lymphocytes from stem cells, Notch signaling directly controls c-Myc expression. Overexpression of active c-Myc promotes while dominant-negative (dn) c-Myc inhibits early cell differentiation. Moreover, c-Myc expression mediated by Notch signaling modulates survivin, an inhibitor of apoptosis (IAP) protein, which is crucial for T cell development. Overexpression of active c-Myc increases while dn c-Myc reduces survivin expression, which is correlated with T cell differentiation within the in vitro differentiation system. These data identify c-Myc, together with survivin, as regulators of the differentiation of T cells from Notch signaling.
Pre-operative immune signatures correlate with recovery from surgical trauma (HUM1P.267)
Authors: Gaudilliere Brice , Fragiadakis Gabriela , Ganio Edward , Tingle Martha , Lancero Hope , Nolan Garry , Angst Martin 
Published: January 2023

Abstract
Traumatic injury produces a profound immune response that mobilizes both innate and adaptive branches of the immune system. Patient recovery after a major trauma is highly variable. Protracted recovery after surgery -a major trauma- causes significant societal and economic costs. However, the mechanistic underpinning of recovery after surgery remains poorly understood. We recently applied high-parameter mass cytometry at the bedside for the deep immune profiling of patients undergoing orthopedic surgery1. The data revealed patient-specific immune responses in monocyte subsets that contained strong correlates of clinical recovery. In this study, we asked whether differences in patients’ pre-surgical immune states determined recovery. Activation of canonical signaling pathways was elicited in vitro, in pre-surgical patient samples. Single-cell evoked immune responses were analyzed in cell subsets spanning the entire immune system using mass cytometry. Surprisingly, the magnitude of these responses varied significantly across patients, thereby defining patient-specific “immune states”. Among these responses, signaling downstream of the TLR4 receptor in CD14+ monocytes strongly correlated with surgical recovery (R=0.64-0.69). The findings highlight a fundamental role for monocytes in the recovery process and provide the mechanistic basis for a pre-operative diagnostic test to predict the course of surgical recovery. 1.Gaudilliere et al., Sci Transl Med. 2014
Decline in relative plasma cell-free mitochondrial DNA levels in response to prolonged moderate aerobic and eccentric exercise (HUM1P.306)
Authors: Shockett Penny , Khanal Januka , Sitaula Alina , Kraemer Robert 
Published: January 2023

Abstract
Elevated plasma levels of cell-free mitochondrial DNA (cf-mDNA), a cell damage-associated molecular pattern (DAMP), contribute to neutrophil activation and inflammation in trauma patients, and may occur in cancer and autoimmunity. To further understand relationships between cf-mDNA released by tissue injury, inflammation, and health benefits of exercise, we conducted trial by time experiments for plasma cf-mDNA response to prolonged moderate aerobic exercise, and to eccentric exercise. Seven healthy moderately-trained young men (n=7) completed treadmill exercise trials for 90 min at 60% VO2 max, and resting control trials, and blood was sampled immediately prior to (Pre), during (at +18, +54, and +90 min), and after (R40) trials. A significant difference in cf-mDNA response was observed between exercise and control trials. Specifically, cf-mDNA levels differed at +54 and +90 (with or without plasma volume (PV) shift correction). A significant time effect was observed, with relative cf-mDNA levels declining at +54 and +90 during exercise. These results suggest increased clearance or reduced release of plasma cf-mDNA with exercise. Declines in cf-mDNA were also seen after eccentric exercise. Our finding of cf-mDNA decline with prolonged moderate treadmill and eccentric exercise contrasts with studies by others involving exhaustive short-term treadmill exercise, where cf-mDNA levels were unchanged, and highlights differences between exercise- and trauma-induced inflammation.
CD4+ T cells from fibrotic livers direct aberrant autoantibody production from B cells (HUM1P.308)
Authors: Tedesco Dana , Thapa Manoj , Mendel Justin , Best Victoria , Ibegbu Chris , Grakoui Arash 
Published: January 2023

Abstract
Two of the most common causes of end-stage liver disease (ESLD) are hepatitis C virus (HCV) infection and alcoholic liver disease (ALD); the only therapeutic option for liver failure is a liver transplant. Extrahepatic manifestations of ESLD include antibody-mediated autoimmunity; an indication of B cell and CD4+ T cell dysfunction. To investigate the contribution of CD4+ T cells during liver disease, we induced liver fibrosis in mice by a conventional weekly carbon-tetrachloride treatment regimen. Fibrotic animals show elevated serum IgG, positive ANA titers and spontaneous IgG production from hepatic B cells. However, this was not evident in CD4+ T cell depleted fibrotic animals despite comparable fibrosis by pathology. Despite significant expression of Foxp3 on fibrotic liver CD4+ T cells, concomitant expression of CD27, PD-1 and CD40L may account for their ability to promote B cell activation. To understand the implications during ESLD in humans, we performed parallel analysis from human liver explant samples. In accordance with our fibrotic mouse model, we saw an enrichment of Foxp3+ CD4+ T cells as well as spontaneous IgG production that was not evident in healthy liver or PBMC. Taken together, these data suggest that altered hepatic CD4+ T cells aberrantly instruct B cells to drive extrahepatic sequelae of ESLD.
Maresin-1 regulates type 2 innate lymphoid cells and promotes generation of regulatory T cells to limit allergic inflammation (HYP2P.336)
Authors: Krishnamoorthy Nandini , Burkett Patrick , Dalli Jessmond , Abdulnour Raja-Elie , Colas Romain , Ramon Sesquile , Phipps Richard , Petasis Nicos , Kuchroo Vijay , Serhan Charles , Levy Bruce 
Published: January 2023

Abstract
Asthma is a chronic inflammatory disease that fails to resolve and is classically considered to be mediated by an overzealous CD4+Th2 immune response. Recently, a key role for type 2 innate lymphoid cells (ILC2s) was shown to have an important role in the initiation and amplification of allergic inflammation; however, mechanisms for ILC2 regulation remain to be determined. In this study, metabololipidomics of murine lungs identified temporal changes in the endogenous levels of docosahexaenoic acid (DHA)-derived pro-resolving mediator, maresin 1, (MaR1) during self-limited allergic inflammation to ovalbumin (OVA). MaR1 decreased after allergen challenge relative to levels found during sensitization, and subsequently increased during the resolution of inflammation. Exogenous administration of MaR1 reduced lung inflammation and ILC2 expression of IL-5 and IL-13 and increased amphiregulin. MaR1 augmented de novo generation of regulatory T cells (Tregs), which interacted with ILC2s to markedly suppress cytokine production in a TGF-β-dependent manner. Antibody-mediated depletion of Tregs interrupted MaR1 control of ILC2 expression of IL-13 in vivo. These results point to a pivotal role for Tregs in the control of ILC2s in a new proresolving mechanism engaged by MaR1.
Bee venom phospholipase A2 suppresses allergic airway inflammation in an ovalbumin-induced asthma model through the recruitment of regulatory T cells (HYP2P.325)
Authors: Bae Hyunsu , Baek Hyunjung , Park Soojin , Jung Kyung-Hwa , Shin Dasom 
Published: January 2023

Abstract
Bee venom (BV) is one of the alternative medicines that have been widely used in the treatment of chronic inflammatory diseases. We previously demonstrated that BV induces immune tolerance by increasing the population of regulatory T cells (Tregs) in immune disorders. However, the major component and how it regulates the immune response have not been elucidated. Here, we investigated whether bee venom phospholipase A2 (bvPLA2) exerts protective effects that are mediated via Tregs in OVA-induced asthma model. The bvPLA2 treatment induced the Tregs and reduced the infiltration of inflammatory cells into the lung in the OVA-challenged mice. Th2 cytokines in the bronchoalveolar lavage fluid (BALF) were reduced in bvPLA2-treated mice. Although bvPLA2 suppressed the number of inflammatory cells after OVA challenge, these effects were not observed in Treg-depleted mice. In addition, we investigated the involvement of CD206 in bvPLA2-mediated immune tolerance in OVA-induced asthma model. We observed a significant reduction in the levels of Th2 cytokines and inflammatory cells in the BALF of bvPLA2-treated OVA-induced mice but not in bvPLA2-treated OVA-induced CD206-/- mice. These results demonstrated that bvPLA2 can mitigate airway inflammation by the recruitment of Tregs in an OVA-induced asthma model.
Novel mechanisms of conferring broad protection against influenza by Lactobacillus casei lactic acid bacteria by mobilizing innate and adaptive immune cells (INC1P.354)
Authors: Jung Yu-Jin , Lee Young-tae , Ko Eun-Ju , Jang Ji-Hun , Park Min-Kyung , Kim Cheol-Hyun , Kang Sang 
Published: January 2023

Abstract
Heat-killed Lactobacillus casei strain lactic acid bacteria DK128 was investigated as potential probiotics to determine its antiviral effects on influenza virus. Intranasal pretreatment of mice with heat-killed DK128 resulted in the recruitment of various innate immune cells into the airways, conferring broad protection against lethal infection with H1, H3, and H5 subtype influenza viruses, preventing body weight loss, and lowering lung viral loads. The protection against lethal infection as a result of killed-DK128 pretreatment was correlated with significantly reduced infiltrates of natural killer cells, neutrophils, and monocytes as well as with increased alveolar macrophage populations. Mice with killed-DK128-mediated protection against H3N2 primary infection effectively developed heterosubtypic adaptive immunity to H1N1 pandemic virus. Further mechanistic studies using various knockout mouse models discovered that B cells but not CD4 and CD8 T cells were absolutely required for heat-killed DK128-mediated protection. Therefore, this study provides evidence that heat-killed DK128 can be developed as beneficial probiotics and that the cross-talk between innate and adaptive immune cells is playing an essential role in conferring influenza antiviral immunity.
Angiopoietins and angiopoietin-like proteins bind to LILRA/B receptors and activate innate immune responses in human monocytes (INC1P.405)
Authors: Person Anthony , Xiong Wendy , Erickson Christian , Shields Kate , Li Jun , BI Ming , Wu Guoping , Kalabokis Vassili 
Published: January 2023

Abstract
Angiopoietins and Angiopoietin-like (ANGPTL) proteins are secreted glycoproteins that play multiple roles in angiogenesis, lipid metabolism, hematopoietic stem cell expansion, and inflammation. Unlike Angiopoietins, which are known to signal through the tyrosine kinase Tie1 or Tie2 receptors, ANGPTL proteins have long been considered orphan ligands. Recently, ANGPTL1, 2, 5, and 7 were shown to bind to human leukocyte immunoglobulin like receptor B2 (LILRB2), resulting in enhancement of human haematopoietic stem cell proliferation and expansion. Leukocyte immunoglobulin-like receptors (LILRs) also named ILTs, LIRs and CD85s, are a family of immunomodulatory molecules that are expressed on professional APCs and regulate their functional properties to influence immune activation and inhibition. While the MHC class I complexes have been identified as the physiological ligands for many LILR members, the direct interaction between LILRs and ANGPTLs has suggested a new signaling mechanism for those two groups of molecules. We have tested Angiopoietins and ANGPTL proteins in binding studies with the LILRA and LILRB family members, and identified additional LILR family members as potential receptors of Angiopoietin-1 and several ANGPTL proteins. We find correlations between these newly described interactions of Angiopoietins and ANGPTL proteins with LILR family members and their abilities to activate innate immune responses in human monocytes.
Immunophenotyping of bone marrow-derived eosinophils in respiratory pathogen infections (INC2P.421)
Authors: Samarasinghe Amali , LeMessurier Kim 
Published: January 2023

Abstract
Asthmatic individuals are considered at high risk of developing viral and bacterial infections. Since eosinophils are abundant in the airways of allergic asthmatics, we hypothesized eosinophils were susceptible to infection by common respiratory pathogens associated with pneumonia. We used bone marrow-derived eosinophils (BMdEos) to simulate exposure to Influenza A Virus (IAV) or Streptococcus pneumoniae. A subset of BMdEos and lung eosinophils exposed to IAV possessed surface-bound nucleoprotein when examined by flow cytometry, indicating both sources of eosinophils were infected with virus. BMdEos also upregulated MHCI and CD86, while MHCII and CD80 remained unchanged. Although microscopically BMdEos appeared homogeneous, two distinct populations based on Siglec-F expression were observed when phenotyped by flow cytometry. MHCI was upregulated almost exclusively by the Siglec-Fhi cells after IAV exposure. Conversely, BMdEos pulsed with S. pneumoniae upregulated MHCII, while downregulating MHCI. Although elevated MHCII was observed for both Siglec-F populations, Siglec-Fint cells upregulated MHCII levels when pulsed with fewer bacteria. Here, we showed that BMdEos can interact with both IAV and S. pneumoniae, responding divergently by modulating surface levels of MHC molecules. Furthermore, we propose the use of BMdEos as a viable in vitro system for examining eosinophils responses to respiratory pathogens.
Effects of nicotine on expression of membrane cofactor protein CD46 in human ARPE-19 cells (INC2P.409)
Authors: Rudolf Xeniya , Lyzogubov Valeriy , Bora Puran , Bora Nalini 
Published: January 2023

Abstract
Age-related macular degeneration (AMD) causes vision loss in humans and its development is often associated with malfunction of the complement system. The complement system is regulated by various proteins including a cofactor protein CD46. Cigarette smoke is a major risk factor for AMD but the impact of nicotine, main component of smoke, on retinal pigment epithelium (RPE) cells is not well studied. We hypothesized that nicotine decreases expression of CD46 in human adult RPE (hARPE-19) cells in vitro. The effects of nicotine were evaluated in two experiments using hARPE-19 cells. In the first experiment, hARPE-19 cells were treated with either 1 µM or 100 µM nicotine. In the second experiment hARPE-19 cells were treated with combinations of 100 µM nicotine and complement depleted or sufficient 10% normal human serum. Western blot and immunofluorescent analyses were used to evaluate the level of CD46 and its cellular localization in hARPE-19. Three isoforms of CD46 occurred in all experimental groups. We suggest that hARPE-19 cells produce full isoform (52 kDa) and two soluble isoforms (38 kDa and 45 kDa). The level of total CD46 and each of the isoforms were similar in all experimental groups. We did not detect any effects of nicotine on the expression of CD46 isoforms with or without active complement system in the medium. Our findings suggest that expression and localization of CD46 on hARPE-19 cells is independent of complement activity, nicotine, or their combination.
Genetic deletion of AU-rich region in the tristetraprolin (TTP) transcript increases TTP stability and expression and provides protection against inflammatory diseases (INM1P.433)
Authors: Patial Sonika , Stumpo Deborah , Huang Weichun , Flake Gordon , Blackshear Perry 
Published: January 2023

Abstract
Tristetraprolin (TTP) is an mRNA binding protein that binds to AU-rich elements (AREs) in the 3’-untranslated regions (3’UTR) of specific mRNAs, such as that encoding tumor necrosis factor (TNF), and increases their rate of mRNA turnover. TTP mRNA also contains AREs within its own 3’UTR that are partly responsible for its own rapid turnover. In the present study, we generated a novel TTP mouse line (TTPΔARE) with a 136 base deletion in the 3’UTR AU-rich region of TTP mRNA, and studied the effect of this genetic modification on the levels of TTP mRNA and protein, and susceptibility of the mice to experimentally-induced inflammatory conditions. TTP mRNA stability was found to be enhanced in TTPΔARE bone marrow-derived macrophages (BMDMs) and mouse embryonic fibroblasts (MEFs). TTP protein expression was increased, both in the basal state and upon LPS stimulation, in BMDMs, and in selected mouse tissues. LPS-induced expression of TTP target mRNAs, particularly Tnf and Il-1β, was significantly reduced in TTPΔARE BMDMs. Most importantly, TTPΔARE mice were almost completely protected from collagen antibody-induced arthritis, and exhibited significantly reduced inflammation in an imiquimod-induced model of psoriasis. These data suggest that increased levels of TTP can protect against some experimentally induced inflammatory diseases. They also raise the possibility that increasing TTP levels in man might be an attractive therapeutic approach in some inflammatory conditions.
Translation inhibition by bacterial pathogens: links to inflammation (INM1P.428)
Authors: Barry Kevin , Fontana Mary , Portman Jonathan , Ingolia Nicholas , Vance Russell 
Published: January 2023

Abstract
Legionella pneumophila is an intracellular bacterial pathogen that is the cause of a severe pneumonia in humans called Legionnaires’ Disease. L. pneumophila infects alveolar macrophages and its virulence requires a type IV secretion system (T4SS) that injects bacterial effector proteins into the host cytosol. Macrophages distinguish infection with T4SS+ (virulent) and T4SS- (avirulent) bacteria, and mount several distinct immune responses to virulent bacteria. Virulent L. pneumophila induces a block in host protein synthesis that is partially induced by 7 bacterial effectors that directly block host translation. L. pneumophila infection also induces the inflammatory cytokine IL-1α. We link translation inhibition to the production of IL-1α, and show that IL-1α signaling, through the interleukin-1 receptor, protects the host during L. pneumophila infection. We show that translation inhibition leads to a sustained production of Il1a transcript and we suggest that this overcomes the block in translation and allows for preferential production of inflammatory cytokines in infected macrophages. To gain insight into the L. pneumophila induced block in host translation, we used ribosome profiling and RNAseq of L. pneumophila infected macrophages to analyze global and mRNA-specific translation rates. Our data suggest that translation inhibition induced by pathogenic infection may be an important pathway that induces inflammation and protects animals from multiple bacterial pathogens.