bluuebunny/crossref_metadata_2025_split huggingface dataset:
Initiation of splenic white pulp ontogeny (HEM3P.289)
Authors: Neely Harold , Flajnik Martin 
Published: January 2023

Abstract
Secondary lymphoid organs (SLO) provide the structural framework for the co-concentration of antigen and lymphocytes required for an efficient adaptive immune response. The spleen is the primordial SLO that evolved coincidentally with Ig-/TCR:pMHC-based adaptive immunity. B cells are indispensable for the formation and maintenance of SLO microarchitecture; their expression of lymphotoxin α1β2 (LTα1β2) is required for the LTα1β2:CXCL13 positive feedback loop without which SLO cannot properly form. Despite the spleen’s central role in the evolution of adaptive immunity, neither the initiating event nor the B cell subset compulsory for white pulp (WP) formation has been identified. We therefore sought to identify these initiating components of splenic WP ontogeny. We detected CXCL13 but not LTα1β2 protein expression in the vasculature of late embryonic (E17.5 and beyond) spleen, and its expression was independent of rearranging lymphocytes and TNFα. A substantial influx of transitional B cells occurred within 18 hours of birth (E18.5), but these cells were unresponsive to CXCL13 and do not establish a perivascular splenic niche. At P0.5, B cells acquired CXCL13 responsiveness and aggregated around the splenic vasculature, and CXCL13-responsive B cells were enriched for transitional 2 B cells. Thus, CXCL13 is the initiating component of the CXCL13:LTα1β2 positive feedback loop required for WP ontogeny, and CXCL13-responsive transitional B cells are the initiating B cell subset.
Normal stem and progenitor cell sociology within the leukemic microenvironment (HEM3P.300)
Authors: Welner Robert , Amabile Giovanni , Bararia Deepak , Czibere Akos , Yang Henry , Tenen Daniel 
Published: January 2023

Abstract
Specialized bone marrow (BM) microenvironment niches are crucial for hematopoietic stem and progenitor cell (HSC/HPC) maintenance. We are just starting to learn how the integrity of the niche changes with leukemia, however, the impact on normal HSC/HPC behavior and functionality is yet to be addressed. Therefore, we started by studying the kinetics and differentiation of normal HSC/HPC in mice with Chronic Myeloid Leukemia (CML). Although normal hematopoiesis was increasingly suppressed during the disease progression, the leukemic environment imposed distinct effects on HPC predisposing them toward the myeloid lineage, similar to that of the leukemic population. Meanwhile, the leukemic-exposed normal HSC remained functional on transplantation. Analysis of the microenvironment identified several cytokines were dysregulated in the leukemia, and we found IL-6 to be responsible for many of these bystander responses. These results were similarly validated using BM obtained from CML patient samples. Co-culture of CML BM and human CD34+ HPC resulted in selective proliferation and altered differentiation of the normal primitive progenitors compared to mixed cultures using normal BM. Therefore, our results show how hematologic malignancy can influence the bone marrow, in turn causing disturbances in normal blood cell progenitors. Knowledge of such bystander effects could suggest new therapeutic interventions for cancer prevention and novel therapeutic approach for leukemia patients.
Vimentin is a dominant target of in situ humoral immunity in human lupus tubulointerstitial nephritis (HUM1P.314)
Authors: Clark Marcus , Chang Anthony , Ko Kichul , Liarski Vladimir , Kinloch Andrew 
Published: January 2023

Abstract
In lupus nephritis (LuN), severity of tubulointerstitial inflammation (TII) on renal biopsy predicts subsequent renal failure. In severe TII, infiltrating B and T cells have features of tertiary lymphoid neogenesis including in situ antigen-driven clonal B cell selection. To identify the autoantigen(s) driving selection, we laser captured single CD38+ or Ki-67+ B cell mRNA from seven LuN biopsies. Twenty clonally expanded immunoglobulin heavy and light chain variable region pairs were cloned and expressed as antibodies. Seven more were obtained from an eighth biopsy by flow sorting of CD38+ cells. Immunofluorescence indicated that 13 of 27 antibodies had immunoreactivity with cytoplasmic structures. Only 4 reacted with nuclei and none with dsDNA, Sm or RNP. Interrogation of protein arrays with TII antibodies and immunoprecipitations followed by mass spectroscopy identified a limited number of self-antigens with only vimentin common to both approaches. Eleven of 13 cytoplasmic TII antibodies directly bound vimentin, vimentin was highly expressed by tubulointerstitial infiltrating cells and tested TII antibodies preferentially bound inflamed tubulointerstitum. Finally, high-titers of serum anti-vimentin antibodies were strongly associated with severe TII. These finding suggest that in situ adaptive immunity to vimentin, an antigenic feature of inflammation, feeds forward to worsen TII and renal damage.
Peripheral blood (PB) monocytes from subjects with autism spectrum disorders (ASD) and fluctuating behavioral symptoms following immune insults reveal similar innate immune responses to those observed in pediatric acute onset neuropsychiatric disorders (PANS). (HUM1P.325)
Authors: Jyonouchi Harumi , Geng Lee , Davidow Amy 
Published: January 2023

Abstract
ASD is a behaviorally defined syndrome and encompasses markedly heterogeneous subjects. Some ASD subjects reveal markedly fluctuating behavioral symptoms following immune insults and persistent gastrointestinal (GI) symptoms; categorized as the ASD-inflammatory subtype (ASD-IS) (N=20) in this study. Cyotkine production by purified PB monocytes from ASD-IS subjects in response to TLR2/6, TLR4, TLR7/8, and Dectin 1 agonists were assessed in comparison with cells from control ASD subjects with and without non-IgE mediated food allergy (NFA) (ASD/NFA N=16, ASD/non-NFA children; N=18), non-ASD controls (non-ASD/NFA; N=13, normal controls; N=12, non-ASD/PANS N=18). ASD-IS subjects had no siblings diagnosed with ASD and ASD-IS children are noted to have a higher frequency of recurrent sinopulmonary infection than ASD/non-ASD controls; 8/20 ASD-IS children were diagnosed with antibody deficiency syndrome (ADS). ASD-IS PB monocytes produced higher levels of IL-1β, IL-6, and IL-12 with or without stimuli. In addition, decrease in production of IL-10 and sTNFRII with TLR2/6 and TLR7/8 agonists was more striking than normal control monocytes. Monocytes from ASD/NFA, ASD/non-NFA, or non-ASD/NFA subjects did not reveal such tendencies. However, cytokine production by PANS monocytes revealed similar tendencies. Aberrant innate immune responses detected in PB monocytes may be associated with markedly fluctuating neuropsychiatric symptoms observed in ASD-IS and PANS subjects.
The role of CCL5 in T cell recruitment to the aorta (HUM1P.303)
Authors: LI Jie , Ley Klaus 
Published: January 2023

Abstract
T cell adaptive immunity is involved in the pathogenesis of atherosclerosis. Previously our lab found that APCs present antigens to effector T cells (Teff). This causes local T cell activation and cytokine production, which sustain local inflammation. However, the chemokine(s) that drive T cell homing to the atherosclerotic aorta are not known. By multi-color flow cytometry to analyze chemokine receptor expression on T cells from Apoe-/- aorta, we found that more than 40% of CD4 T cells in Apoe-/- aorta express CCR5. Notably, &gt;80% of the CCR5+ T cells in the plaques are IFN-γ+Foxp3+ Tregs. This is a unique CD4 T cell subset in atherosclerosis. They are found only in the aorta and para-aortic lymph nodes of Apoe-/- mice but not in the spleen or other organs, suggesting that the generation of the Th1-like Tregs is associated with local antigen presentation. To test my hypothesis that CCR5 and its ligand CCL5 play an important role in Teff recruitment to the aorta, Teff cells from CD45.1Apoe-/- mice were incubated with explanted Apoe-/- aortas with media alone, CCL5 or CCR5 neutralizing antibody. The CCL5 and CCR5 neutralizing antibody significantly decreased the number of T cells infiltrating into the aorta (53% and 70% respectively vs. media alone). The result is also confirmed by in vivo adoptive transfer experiment using leukocytes from CCR5-/-Apoe-/-mice. CCR5-deficient CD4 T cells displayed dramatic decrease in homing to the aorta compared to the CCR5 competent T cells.
TREML4 expression by myeloid cells may play a role in coronary artery disease (HUM1P.308)
Authors: Boelte Kimberly , Sen Shurjo , Long Priel Debra , Lau Karen , Cardone Marco , Yoshimura Teizo , Barb Jennifer , Joehanes Roby , Accame David , Teer Jamie , Singh Larry , Adams Lila , Zhang XiaoQing , Chowdhury Soma , Johnson Andrew , Green Eric , Mullikin James , Kolodgie Frank , Virmani Renu , O'Donnell Christopher , Munson Peter , Trinchieri Giorgio , Kuhns Douglas , Biesecker Leslie , McVicar Daniel 
Published: January 2023

Abstract
Triggering Receptor Expressed on Myeloid cells (TREM) family receptors modulate responses of a variety of leukocytes. We found that TREM-like 4 (TREML4) mRNA is elevated in the peripheral blood of patients with advanced coronary artery calcification (CAC) and that a single nucleotide polymorphism (SNP), rs2803496, tracks with the level of TREML4 mRNA in these patients. In an effort to elucidate the mechanism through which TREML4 in peripheral blood could affect CAC, we began by determining what cells express it. Monocytes and lymphocytes of donors with this SNP expressed very low levels of TREML4 mRNA whereas their neutrophils (PMNs) were strongly positive. Consistent with its association with CAC, we found variation in the levels of TREML4 mRNA in PMNs that correlated rs2803496, and a second SNP rs2803495. Unlike murine Treml4, which has the typical TREM family structure, human TREML4 has a noncanonical leader, lacks a portion of the transmembrane domain (TM) and has no cytoplasmic tail. Characterization of the Treml4 cDNA suggests that this truncated TM domain is sufficient to anchor the protein in the plasma membrane but that the TREML4 native leader sequence is ineffective. Lastly, immunostaining of human atherosclerotic plaques shows TREML4 in macrophage-rich regions, as well as on the endothelium. Together our findings suggest that TREML4 may play an important role in atherosclerosis and CAC but that it is unlikely to function as a typical, DAP12-coupled TREM.
A defect of differentiation of peripheral blood monocytes into dendritic cells after cardiopulmonary bypass is delayed by 7 days after the initial insult (HUM1P.309)
Authors: Laudanski Krzysztof , Worthen G. Scott 
Published: January 2023

Abstract
Monocytes (MO) can uniquely differentiate in a variety of the offspring cells including dendritic cells (DC). After severe immune insult, like sepsis, ability of MO to differentiate to DC (MO-&gt;DC) is significantly diminished immediately after insult and is related to the acquisition of a predominantly macrophage like characteristic of MO thus leaving only few available precursory MO for MO-&gt;DC process. Here, we enrolled patients scheduled for non-emergent heart surgery and assessed leukocyte function before surgery (t0), 24hrs (t24h) and 7 days (t7d) after surgery. We hypothesized that immediately after surgery, MO will acquire a persistent macrophage characteristic with subsequent MO-&gt;DC defect. MO were isolated and subjected to a DC differentiation&maturation conditions including 5 days in vitro cultures in X-VIVO15 enriched with IL-4 & GM-CSF. The process of MO-&gt;DC was significantly diminished in patients but only at the day 7 after the CPB as shown by diminished expression CD83 (%CD83+ [X±SD] t0=24.5±33.81 vs t24hr=8.84±10.71 [p=ns] vs t7d=0.2±0.4 [p&lt;0.05]) or uptake of DQ ovalbumin. Concomitantly, the number of precursory CD11c+ was diminished but only at 7 days samples (%CD11c+ [X±SD] t0=81.5±15.81 vs t24hr=83.3±15.82 [p=ns] vs t7d=61.9±18.32 [p&lt;0.05]). We conclude that surprisingly CPB diminished number of circulating precursory DC but only after 7 days from insult affecting ability of these precursory cells to convert into DC.
Regulation of microRNA expression in the lungs by exposure to air pollution & antigen (HUM2P.340)
Authors: Lucas Benjamin , Chen Wen-Chi , Bleck Bertram , Reibman Joan , Rom William , Narayanan Narayanan , Qu Qingshan , Park Sung-Hyun , Grunig Gabriele 
Published: January 2023

Abstract
Pulmonary Hypertension is characterized by pulmonary arterial remodeling and increased pressure in the pulmonary circulation. It is often associated with inflammation in the lungs and can lead to right heart failure. Our work shows that urban ambient pollution exacerbates the experimental pulmonary hypertension phenotype just like other types of inflammatory lung conditions. We aimed to identify microRNAs (miRNAs) that are differentially expressed in our mouse model. In addition, we examined plasma samples from individuals occupationally exposed to high levels of air pollution or cigarette smoke, and from controls. Our study is the first to show significantly de-regulated expression of three microRNA species (miR-135a, miR-21, miR-204) in the lungs of mice that were exposed to antigen and particulate matter and developed pulmonary hypertension. De-regulated levels of miR-21 and miR-204 have been reported in human pulmonary hypertension and in experimental pulmonary hypertension. MiR-135a is targeting STAT6 and upregulated expression has been reported in experimental asthma. Using human samples, our study showed that plasma levels of miR-21 and miR-135a, but not levels of miR-204, clustered individuals with high dose exposures and individuals with low dose environmental exposures. Current studies are aimed at identifying the cytokines that control these miRNAs’ expression. The long range goal is to identify miRNAs that indicate an at-risk state of the pulmonary vasculature.
Protoarray analysis reveals novel autoantigens targeted by autoantibodies associated with DNA-repair pathway in systemic erythematosus lupus (HUM2P.330)
Authors: Li Quanzhen , Zhou Jinchun , Wu Tianfu , Mohan Chandra 
Published: January 2023

Abstract
Systemic erythematosus lupus (SLE) is an autoimmune disease characterized by presence of autoantibodies (autoAbs) against a broad spectrum of self-antigens. In order to identify novel autoAbs associated with SLE, we utilized a Protoarray bearing 9,500 antigens to screen IgG and IgM autoAbs in sera of SLE patients. 446 IgG and 1218 IgM autoAbs were identified to be significantly elevated in SLE patients compared with healthy controls (p&lt;0.05). Protoarray revealed not only the previously described autoAbs such as antibodies against dsDNA, SSA/SSB, Histone and Sm/RNP, but also uncovered autoAbs against a broad range of novel antigens including 151 nuclear-associated antigens and 150 cytoplasma or membrane-associated antigens. The Protoarray results were further validated by ELISA in a larger cohort of SLE patients and controls. Besides, 65 of the IgG autoAb-targeted proteins were also disregulated in SLE on mRNA level by microarray analysis. Pathway analysis recognized significant enrichment of antigens involved in cell proliferation and DNA repair pathways which were targeted by IgG autoAbs including APEX1, AURKA, CSNK1G1, EIF2C1, HMGB1, IFIT5, MAPKAPK3, PADI4, PRKRA, RALGPS1, UBE2S and VRK1. The elevated IgG autoAbs to APEX1 and other proteins in the pathway may reflect the compromised DNA-repair activity during DNA replication in SLE. In conclusion, identification of novel autoAbs by protoarray may shed light on some of the pathogenic pathways leading to disease in SLE.
Genetic risk alleles associated with autoantibody production and responsible for initial breach in tolerance to self-antigens in normal individuals (HUM2P.331)
Authors: Li Quanzhen , Raj Prithvi , Raman Indu , Yan Mei , Lian Yun , Zhang Bo , Wakeland Edward 
Published: January 2023

Abstract
Antinuclear antibodies (ANA) are serological hallmarks of SLE. Studies have shown that healthy individuals also develop ANA, raising questions on relevance of ANA positivity and autoimmune disease. We screened 2223 healthy controls (HC) and 143 SLE patients for ANA by ELISA and found that 26% HC are ANA positive, with 17% moderate (20-40 AU) and 9% high (&gt;40 AU) ANA levels. To elucidate the autoantibody profiles associated with ANA in HC and SLE, we performed autoantigen arrays to profile over 90 autoAbs and found that ANA positive HC make significant amount of IgG and IgM autoAbs against many antigens with high frequency to non-nuclear antigens such as collagens and phospholipids. We noticed stronger autoAb signatures in females than males. We then genotyped 1583 HC with high and low ANA by Immunochip and did quantitative association test using ANA as dependent variable. Seven genomic loci showed suggestive genomewide association (p=1.00E-05) with ANA. Multiple independent association signals were observed in HLA class I, II and III region and the strongest association mapped to HLA-DR alleles where peak SNP rs9268832 in HLA-DRA gene reached p value 7.63606E-07. The impact of ANA risk alleles on HLA-DR gene expression was studied in B cells and macrophages from healthy donors by RNA-seq and results show that ANA risk haplotype is associated with up regulation of HLA-DRB1 gene which could be associated with initial breach in tolerance to self-antigens in general population.
MS4A4 regulates KIT receptor trafficking, signaling and function in mast cells (HYP3P.343)
Authors: Cruse Glenn , Music Stephen , Bradding Peter , Beaven Michael , Metcalfe Dean 
Published: January 2023

Abstract
Expression of MS4A family proteins influence the cell cycle and may be involved in tumorigenesis. However, the mechanistic basis for these observations remains elusive. We have identified roles for a member of the MS4A family, FcεRIβ (MS4A2), in protein trafficking and signal transduction in human mast cells (Immunity, 38, p906, 2013) and predicted analogous roles for other MS4A proteins. Employing mast cells, we found that down-regulation of MS4A4 reduced KIT expression (the receptor for SCF), and contributed to effector functions. MS4A4 “stabilized” surface KIT by slowing KIT internalization kinetics in response to SCF. Our data suggest that the mechanism for this action may involve trafficking of KIT by MS4A4 from sorting early endosomes to the endocytic recycling center, diverting receptors away from late endosome-dependent ERK kinase signaling and receptor degradation. Thus, in this instance, gene-silencing of MS4A4 reduced recycling pathways resulting in enlarged EEA1+ early endosomes, prolonged ERK signaling, and increased KIT progression to Rab9+ late endosomes. Therefore, MS4A4 appears to promote mast cell maturation by altering the endocytic trafficking and localization of KIT signaling, fine tuning effector functions. Our proposed function of MS4A4 in KIT trafficking is similar to the role that FcεRIβ plays in FcεRI trafficking and raises the question of whether other MS4A gene family members similarly regulate trafficking of receptors and associated molecules.
Complete suppression of IgE-mediated anaphylaxis by antihistamine plus tyrosine kinase antagonists. (HYP3P.400)
Authors: Morris Suzanne , Perkins Charles , Khodoun Marat , Hogan Simon P , Finkelman Fred 
Published: January 2023

Abstract
IgE/FcεRI-mediated mast cell and basophil activation has a central role in food allergy and drug allergy. Although desensitization therapy can suppress these disorders, it can be complicated by anaphylaxis. Pre-treatment with antihistamines and other mediator antagonists can increase the safety of desensitization, but is not completely effective. Because the tyrosine kinase Syk has an important role in FcεRI-dependent mast cell activation and the tyrosine kinase Kit is critical for mast cell survival, we evaluated whether treating mice systemically with a combination of an antihistamine (triprolidine, 200 µg), the Syk inhibitor fostamatinib (80 mg/kg), and the c-Abl/c-Kit inhibitor, imatinib mesylate (1.25 mg), 30 min prior to challenging them intravenously with an anti-IgE mAb, would prevent development of anaphylaxis (measured by hypothermia); mast cell degranulation (measured by increases in serum MMCP1); and/or basophil activation (measured by increases in IL-4 secretion). Pretreatment with this 3-agent cocktail completely suppressed hypothermia and decreased MMCP1 by &gt;80%, but had little effect on IL-4 secretion. Triprolidine and fostamatinib independently partially suppressed hypothermia and fostamatinib, by itself, suppressed MMCP1 by 50-60%. No obvious drug toxicity was noted. These results suggest the possibility of using a combination of mediator and tyrosine kinase antagonists to increase the safety of rapid and conventional desensitization therapy.
n-3 polyunsaturated fatty acids (PUFAs) inhibit mast cell activation by disrupting FcεRI association with lipid rafts (HYP3P.401)
Authors: Wang Xiaofeng , Kulka Marianna 
Published: January 2023

Abstract
INTRODUCTION: n-3 PUFAs can suppress allergic inflammation. We hypothesized that n-3 PUFAs inhibit mast cell (MC) activation through modification of FcεRI association with lipid rafts on plasma membrane. METHODS: Bone marrow-derived MCs (BMMC) were cultivated from C57BL/6 wild-type (WT) and fat-1 transgenic mice which express fatty acid n-3 desaturase and produce endogenous n-3 PUFAs. Exogenous n-3 PUFAs were supplemented to WT BMMC. Degranulation, cysteinyl leukotriene (cys-LT), and tumor necrosis factor (TNF) and chemokine (C-C motif) ligand 2 (CCL2) release were evaluated following FcεRI activation. Lipid rafts were isolated by sucrose gradient centrifugation. FcεRI expression was determined by flow cytometry and western blot analysis. RESULTS: Gas chromatography-mass spectrometry analysis showed increased n-3 PUFA levels in whole cell lysates and lipid raft fractions of fat-1 BMMC compared to WT BMMC and in whole cell lysates of PUFA-supplemented BMMC. n-3 PUFAs inhibited degranulation and cys-LT, TNF and CCL2 production compared to untreated BMMC. n-3 PUFAs did not alter expression of surface or total cell FcεRI. However, n-3 PUFAs suppressed FcεRI localization in lipid rafts of unstimulated cells, and disrupted FcεRI shuttling to rafts in stimulated BMMC. CONCLUSIONS: Our results suggest that n-3 PUFAs inhibit MC activation by disruption of FcεRI localization and shuttling into lipid rafts.
The role of lactic acid on mast cell function (HYP3P.404)
Authors: Spence Andrew , Abebayehu Daniel , Ryan John 
Published: January 2023

Abstract
Lactic acid is produced from pyruvate during anaerobic respiration due to a high demand and low supply of oxygen. Therefore, it stands to reason that hypoxic tissues such as tumors or asthmatic bronchioles would have increased lactic acid production relative to their healthy counterparts. Mast cells are also prevalent in these tissues. The purpose of this research is to determine the effect lactic acid can have on mast cell function. Murine bone marrow derived mast cells were cultured in the presence of various amounts of L-(+)-lactic acid (0-25mM). BMMCs were activated with either IgE+antigen or IL-33. Cytokine and chemokine production was measured using ELISAs. Our results showed that IgE-mediated cytokine/chemokine production increased with the amount of lactic acid, with significantly enhanced MCP-1, IL-6 and TNFα at 12.5mM lactic acid. However, IL-33-induced cytokine/chemokine production was decreased in the presence of lactic acid, with significant suppression of MCP-1 and IL-6 at 12.5mM lactic acid. These data suggest that a hypoxic environment can alter mast cell inflammatory responses in a stimulus-dependent manner, and warrant further study into molecular mechanisms.