bluuebunny/crossref_metadata_2025_split huggingface dataset:
The cysteinyl leukotriene receptor CysLTR1 mediates Th17 cell migration (CAM4P.156)
Authors: Kim Hyeong Su , Lee Gap 
Published: January 2023

Abstract
Th17 cells are one of CD4+ T cells that have crucial role in inflammation and autoimmunity. Although many studies revealed molecular mechanisms about Th17 cells but the migration of Th17 cells by lipid-based chemoattractant leukotriene signaling is not fully understood. Here, we show that Th17 cells are highly sensitive to leukotriene B4 and D4. Th17 cells expressed high levels of leukotriene B4 receptors and cysteinyl leukotriene receptors compare to other effector CD4+ T cell subsets. In chemotaxis assay, we confirmed that migration of Th17 cells was more efficient than that of other effector T CD4+ T cell subsets under the guidance of leukotriene B4 and D4, and it was blocked by specific inhibitors of leukotriene B4 receptor and cysteinyl leukotriene receptor. Finally in an animal disease model of experimental autoimmune encephalomyelitis (EAE), treatment with montelukast which is an inhibitor of cysteinyl leukotriene receptor, it mitigated clinical score of EAE. Thus, we concluded that leukotrienes act as chemoattractant for Th17 cells.
Antibody blocking of CLEC12A delays the course and attenuates EAE severity by impairing myeloid cell CNS infiltration: A potential new therapy for multiple sclerosis (CAM4P.158)
Authors: Sagar Divya , Singh Narendra , Huang Xiaofang , Ginwala Rashida , Philip Ramila , Nagarkatti Mitzi , Nagarkatti Prakash , Khan Zafar , Jain Pooja 
Published: January 2023

Abstract
The mechanism of dendritic cell (DC) recruitment across the blood brain barrier during neuroinflammation has been the least explored amongst all leukocytes. For cells of myeloid origin, while integrins function at the level of adhesion, the importance of lectins remains unknown. We pursued the study of a highly specific aspect of DC biology, namely the expression of surface C-type lectin receptors (CLRs) for their importance in adherence and transmigration across the blood-brain barrier in response to CCL2 chemotaxis. CLEC12A, an immunoreceptor tyrosine-based inhibitory motif (ITIM) known to recruit Src homology region 2 domain-containing phosphatase (SHP)-1/2, was subsequently identified as one such a target on DCs. SHP-1/2 phosphorylation was further found to facilitate the actin polymerization on DCs that express the WASP Interacting Protein (WIP). To test function of CLEC12A in an animal model of multiple sclerosis (MS), we administered an antibody to block CLEC12A that significantly ameliorated the progression and relapse of experimental autoimmune encephalomyelitis (EAE) in mice through inhibition of demyelination and myeloid cell infiltration into the CNS tissue. Correspondingly, a restoration of DC numbers in the periphery along with decrease in TH17 phenotype within CD4+ T cells was also seen. These studies reveal the utility of a DC-specific mechanism in designing new therapeutics for MS.
The recruitment of macrophages to the pre-ovulatory ovary (CAM4P.146)
Authors: Whitaker Alex , Linville Elizabeth , Oakley Oliver 
Published: January 2023

Abstract
Ovulation requires the chemotaxis of leukocytes from the spleen to the ovary. Hormonal signals that are released during the peri-ovulatory period initiate the release of specific leukocyte populations that infiltrate the ovary and facilitate the release of oocytes. We identified a specific population of monocyte that infiltrates the ovary during a specific period of time, as well as M1 and M2-like macrophage populations. Before ovulation occurs, a Ly-6hi monocyte is likely promoting inflammation and follicular rupture. The Ly-6c high monocyte infiltration will start to diminish as they differentiate into M1 macrophages to further promote inflammation. These studies will utilize a super-ovulation protocol to initiate ovulation in immature balb/c mice. Ovaries will be collected at several times post hCG injection and analyzed by multi-color flow cytometry. Oviducts will be collected from mice 20 hours after hCG administration and cumulous oocyte complexes will be collected, counted and averaged to determine ovulation rate. Studies should establish that without these subsets of monocytes and macrophages, the rate of ovulation is reduced. In this regard, in the ovary, the Ly-6c high monocytes may function as a source of M1 macrophages to initiate the inflammatory process.
Roles of the CCL20-CCR6 axis and CXCR3 in CD8 T cell-mediated contact hypersensitivity (CCR5P.211)
Authors: Kish Danielle 
Published: January 2023

Abstract
Contact hypersensitivity (CHS) is a CD8 T cell-mediated response to skin sensitization and challenge with hapten. Hapten-primed CD8 T cell populations producing IFN-g and IL-17 are required to induce hapten-presenting endothelial cells (EC) in the challenged skin to produce CXCL1 and CXCL2 that direct Gr-1+CXCR2+ cell recruitment into the skin challenge site during elicitation of CHS. This study investigated mechanisms directing the localization of the hapten-primed IL-17- and IFN-g producing CD8 T cells to EC in the challenge site. qPCR analyses indicate DNFB hapten application to the skin of sensitized mice or to EC cultures induces TNFa-dependent expression of CCL20 within 1 h. Hapten sensitized mice treated with anti-CCL20 mAb at the time of challenge decreases CXCL1 and IL-17, but not IFN-g, expression in skin challenge site 4 hrs. post-challenge. ELISPOT assay analyses of flow-sorted CD8 T cells from draining lymph nodes of sensitized mice indicate populations of CCR6+CXCR3+, CCR6+CXCR3- and CCR6-CXCR3+ cells producing IL-17 in response to hapten-presenting cells. In contrast, IFN-g is produced only by CXCR3+ CCR6- cells. Hapten sensitization and challenge of CCR6-/- mice induces CHS responses that are only slightly lower than those in WT controls. These results indicate a role for CCR6+, CCR6- and CXCR3+ CD8 T cells in producing the IL-17 and IFN-g required for elicitation of CHS responses.
Pathogenic regulatory circuits in B cell lymphoma (HEM8P.238)
Authors: Koues Olivia , Kowalewski Rodney , Schmidt Jennifer , Pyfrom Sarah , Cashen Amanda , Payton Jacqueline , Oltz Eugene 
Published: January 2023

Abstract
Reprogramming gene expression via epigenetic perturbation of transcriptional control elements has emerged as a major mechanism of oncogenesis. Non-Hodgkin Lymphoma (NHL) is the most frequently diagnosed hematopoietic tumor. While distinct NHL gene expression patterns were identified a decade ago, underlying changes to the control elements driving these alterations remain poorly understood. Comparative integration of epigenome and transcriptome data from NHL cells and their normal counterpart B cells reveal the oncogenic regulatory circuitry that links differentially regulated control elements to target genes. Although many of these circuits are engaged in GC B cells or during B cell activation, a portion is commandeered from other cell lineages to promote transformation. Independent sets of transcription factors, whose expression are also deregulated in NHL, target augmented or attenuated circuit components. Importantly, these components are enriched for sequence variants that disrupt transcription factor binding and expression of circuit-linked genes. The pathogenic regulatory circuitry of NHL reveals genetic and epigenetic etiologies for B-cell transformation, providing unprecedented opportunity for therapeutic intervention.
Recombinant Salmonella typhimurium outer membrane protein A and D reactive T cells are expanded in synovial fluid of patients with reactive arthritis and undifferentiated spondyloarthropathy (HUM6P.251)
Authors: Chaurasia Smriti , Shasany Ajit , Aggarwal Amita , Misra Ramnath 
Published: January 2023

Abstract
Introduction: Low molecular weight fractions of outer membrane protein (OMP) of S. typhimurium have been shown to stimulate synovial fluid mononuclear cells (SFMCs) of patients in reactive arthritis (ReA) and undifferentiated spondyloarthropathy (uSpA). By proteomic analysis the stimulatory fractions predicted to consists of 10 proteins including OMPA and OMPD. We studied the frequency of recombinant OMPA and OMPD reactive T cells in the synovial fluid (SF) of patients with ReA/uSpA. Methods: Recombinant OMPA and D were synthesized and SFMCs of 20 patients with ReA/uSpA and 8 patients with rheumatoid arthritis (RA) were cultured with rOMPA/D. IFN-γ and IL17 production by SF T cells was assessed by Flow cytometry and in culture supernatant by ELISA. Antigen specific IFN-γ producing T cells were also enumerated by ELISPOT. A positive response was taken if % cells or Mean spots/well with antigen/Control ≥ 3 Results: Frequency of IFN-γ producing CD8 T cells was increased to crude OMP in 13/20, rOMPA in 9/20 and to rOMPD in 4/20 patients. Frequency of IFN-γ producing CD4 T cells was increased to crude OMP in 6/15, rOMPA in 4/15 and to rOMPD in 2/15 patients. By ELISPOT, 13, 7 and 3 patients showed increased IFN-γ producing SF T cells to crude OMP, rOMPA and rOMPD respectively. IFN-γ and IL17 levels were found to be significantly increased in culture supernatant of cells treated with rOMPA. Conclusion: OMPA is a major antigenic target in patients with Salmonella triggered ReA/uSpA.
Gender differences at baseline in autoantibody levels but no gender differences in vaccine-induced responses and no changes in autoantibody levels following influenza vaccine (HUM6P.246)
Authors: Jiang Wei , Kilby J. , Gilkeson Gary 
Published: January 2023

Abstract
Background. Gender bias presents in most autoimmune diseases such as systemic lupus erythematosus (SLE) disease. In the current study, we investigate the mechanisms of gender bias in autoreactive antibody (Ab) production in healthy controls after influenza (flu) vaccination. Method. Plasma levels of anti-nuclear Ab (ANA), anti-dsDNA autoAb, and anti-CD4 autoAb in a cohort of 5 healthy men and 11 healthy women received vaccines at 2012-2013 and 2013-2014 seasons. Blood draws were taken at 0, 7, and 14 days after the latest vaccination. Vaccine responses were defined by neutralization activities in plasma. Flu-specific Ab avidity was tested by ELISA. The levels of autoAbs were analyzed in plasma by ELISA. Results: Women have higher levels of all 3 autoAbs than men at D0, but not the flu-specific neutralizing activities or flu-specific Ab avidities. The median plasma levels of anti-ANA Ab (OD) at D0 were 0.2281 (IQR, 0.2058 - 0.2338) and 0.3697 (IQR, 0.2918 - 0.4261); the median plasma levels of anti-CD4 Ab (OD) were 0.1678 (IQR, 0.1589 - 0.1932) and 0.2148 (IQR, 0.1986 - 0.2696); and the median plasma levels of anti-dsDNA Ab (IU/mL) were 122.3 (IQR, 91.86 - 175.8) and 197.3 (IQR, 131.2 - 389.8), for men and women respectively (P &lt; 0.05, Mann Whitney U test). Influenza vaccination did not change the titer of autoAbs at any time point. Conclusions: Women have increased levels of autoAbs than men, and these autoAbs are not resulted from repeated influenza vaccinations.
Activation of monocytes through FcRγ-signaling promotes IL-33-dependent migration into the lung interstitium (HYP7P.270)
Authors: Tjota Melissa , Sperling Anne 
Published: January 2023

Abstract
The development of Type 2 responses, such as those seen in asthma, has been significantly associated with IL-33. Previously, we demonstrated that allergen-specific IgG immune complex (IC) signaling through FcγRIII on IL-33 sufficient DCs induced Type 2 allergic airway inflammation. These data suggested that IL-33-/- mice, which had diminished Type 2 responses in the lungs, had a defect in APC function. For this study, we focused on monocytes and CD11b+ monocyte-derived DCs (moDCs) as they were shown to have the highest levels of FcγRIII expression, antigen uptake, and IL-33 upregulation. Notably, there was a significant reduction in the numbers of monocytes and CD11b+ moDCs in the lungs of IL-33-/- mice compared to WT mice after IC sensitization, although no defect was observed at baseline. Using intravascular staining to identify cell localization, we demonstrated that antigen uptake of ICs promoted migration of monocytes from the lung vasculature to the lung interstitium in an FcγRIII- and IL-33-dependent manner. Functionally, failure of monocyte migration during IC-mediated Type 2 inflammation led to reduced eosinophilia in the lungs. Overall, our findings suggest that during sensitization, activation of FcγRIII in monocytes promotes migration into the interstitium in an IL-33 dependent manner, and that this migration is essential for the development of Type 2 responses in the lungs during allergic airway inflammation.
Preparation of human IgG1 monoclonal antibody specific for house dust mite allergens Der p 2 and Der f 2 by single B cell PCR of VH and VL segments (HYP7P.275)
Authors: Chen Jou-Han , Chen Jiun-Bo , Hsieh Yi-Chun , Chu Chia-Yu , Chang Tse-Wen 
Published: January 2023

Abstract
In allergic patients sensitive to house dust mite allergens, the inhaled allergenic proteins are dissolved in mucosal fluid, cross mucosal epithelia of the airway, and induce crosslinking of FcεRI-bound allergen-specific IgE on mast cells and basophils, leading to the degranulation of those inflammatory cells. Our group has proposed that allergen-specific IgG, delivered to the airway mucosa, can bind to the incoming allergens and preclude their entry into the mucosal layer. In this work, we prepared human monoclonal antibodies by isolating dust mite allergen-specific single B cells from patients with atopic dermatitis and performing PCR on the VH and VL cDNA of the B cells. One mAb, 1D11, an IgG1 (γ1, λ), bound strongly to both Der p 2 and Der f 2 and recognized a linear epitope in the peptide segment, a.a. residues #41-70, of the two proteins. We proceeded evaluating the ability of 1D11 to trap Der p 2 and Der f 2 in a culture of rat basophilic leukemic (RBL) SX-38 cells, which were charged with allergen-specific IgE contained in patient sera. In the culture, 1D11 was capable of inhibiting the degranulation of RBL SX-38 cells triggered by Der p 2 and Der f 2. Furthermore, the activation was reduced to the background level when protein A-conjugated Sepharose was employed to remove the 1D11-allergen complexes. These results indicate that mAb 1D11 is effective in trapping Der p 2 and Der f 2 allergens and prevent their activation of allergen-specific IgE-sensitized basophils.
Regulation of neutrophil cytokine production during inflammation (INC6P.319)
Authors: Deguine Jacques , Sinha Meenal , Lowell Clifford , Barton Gregory 
Published: January 2023

Abstract
Neutrophils are usually among the first immune cells present at a site of inflammation, and are critical for bacterial elimination. While neutrophils produce a variety of cytokines and chemokines, the local regulation of this activity, and its contribution to inflammation, remain unclear. To determine whether neutrophil cytokine production is affected by local cues, we studied neutrophils from various inflammatory sites. Here, we show that neutrophils elicited in the presence of ligands for C-type lectin receptors (CLR) can produce high levels of several pro-inflammatory cytokines, while naïve neutrophils or neutrophils elicited in the absence of CLR ligands are poor cytokine producers. Interestingly, this increase is not mediated by a transcriptional increase in cytokine mRNAs, but rather by a direct increase in protein production. Furthermore, we demonstrate that increase of cytokine production does not require direct sensing of CLR ligands, but is instead mediated by paracrine signals delivered by neutrophils themselves. Altogether, our results show that specific local cues influence the ability of neutrophils to produce cytokines. Identifying these signals may provide us with new tools to modulate the functions of neutrophils during infections and inflammatory diseases.
SLO induced inflammasome activation is triggered by pore-formation instead of membrane repair (INM6P.337)
Authors: Romero Matthew , Bhattacharjee Pushpak , Keyel Peter 
Published: January 2023

Abstract
Macrophages activate membrane repair and the inflammasome in response to bacterial pore-forming toxins like streptolysin O (SLO). SLO mutants that are more easily resisted by repair processes produce increased amounts of IL-1β. This suggests that membrane repair may influence inflammasome activation, though it is not clear whether pore-formation or triggering repair pathways directly activates the inflammasome. In order to determine the relationship between membrane repair and the inflammasome, we first determined the mechanism of membrane repair in macrophages. Dose-dependent cytotoxicity of SLO is lower in macrophages than in either 3T3 or HeLa cell lines, indicating increased repair efficiency in macrophages. This was not due to decreased SLO binding to macrophages. Macrophages challenged with sublytic doses of SLO shed microvesicles similar to HeLa and 3T3 cells. Although pore-formation was necessary for inflammasome activation, the pore-dead SLO N402E still promoted microvesicle shedding. However, a second SLO pore-dead mutant, SLO G395V/G396V failed to promote shedding. This suggests that membrane binding and the extent of oligomerization, rather than pore-formation, is necessary to trigger membrane repair. Since pore-formation is necessary for IL-1β secretion, these data suggest that membrane repair itself does not promote inflammasome activation. Instead membrane repair preserves the cell long enough to execute its inflammatory program.
Exploring mechanisms of Interleukin-1α release from HSV-1 infected keratinocytes during active skin infection (INM6P.340)
Authors: Milora Katelynn , Jensen Liselotte 
Published: January 2023

Abstract
IL-1 cytokines, IL-1α and IL-1β, have been widely studied in a variety of disease models. These molecules are mostly known for their inflammatory properties; however, they have been shown to have pleiotropic effects under diverse circumstances and within different cell types. Recently, the herpes simplex virus-1 (HSV-1) was shown to inhibit IL-1β release during infection and consequently possibly blocking IL-1β dependent immune responses. We previously demonstrated that IL-1α is expressed and released from cells exposed to bacteria. Utilizing a mouse model of HSV-1 skin infection, we recently further showed that an intracellular store of preformed IL-1α is released from keratinocytes soon after infection, independent of inflammatory caspase activation. IL-1α signaling through its receptor, IL-1R1, was subsequently responsible for leukocyte recruitment to infected epidermal cells. This showed, for the first time, this cytokine’s ability to act as an alarmin during cutaneous viral infection. Consequently, IL-1R1 knockout mice had an increased mortality rate compared to wild type mice due to viral dissemination. Recent studies have aimed to evaluate mechanisms whereby IL-1α is released from infected keratinocytes and discover potential proteins involved. These experiments will help to elucidate the role of IL-1α during viral infection while furthering our understanding of IL-1 signaling mechanisms.
SHARPIN, a novel regulator of the NLRP3 inflammasome (INM6P.334)
Authors: Gurung Prajwal , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
NLRP3 recruits adaptor ASC and protease caspase-1 to assemble the inflammasome, which is critical for maturation of IL-1β and IL-18. Although, NLRP3 is one of the most-studied inflammasome, the direct ligand or the mechanisms that govern activation of NLRP3 inflammasome remains obscure. Recent study with Hoil1-/- mice demonstrated that HOIL1 is required for NLRP3 inflammasome activation. It was further proposed that HOIL1 was required for linear ubiquitination of ASC to regulate NLRP3 inflammasome assembly. HOIL1 along with HOIP and SHARPIN form linear ubiquitin assembly complex (LUBAC) that is essential for linear ubiquitination of target proteins. Herein, we wanted to determine whether SHARPIN is also involved in the activation of NLRP3 inflammasome. Using Sharpincpdm mice (spontaneous mutation that results in SHARPIN deficiency), we demonstrated that SHARPIN is critically required for activation of both canonical and non-canonical NLRP3 inflammasome, but dispensable for activation of NLRC4 and AIM2 inflammasome. Interestingly, our studies further show that unlike HOIL1, SHARPIN regulates NLRP3 inflammasome by controlling priming events. To this end, we demonstrated that Sharpincpdm BMDMs have dramatically reduced NFkB, ERK, p38 MAP kinase activation and pro-Il1b expression. Thus, our study for the first time demonstrates SHARPIN as a novel regulator of NLRP3 inflammasome adding to the complexity of our understanding of NLRP3 inflammasome.
Global regulation of Toll-like receptor 4-induced inflammatory gene networks by physiologic stress signals in macrophages (INM7P.356)
Authors: Botten Giovanni , Scumpia Philip , Doty Kevin , Smale Stephen 
Published: January 2023

Abstract
Physiologic stress mediated through adrenergic (catecholamines) and neuroendocrine (glucocorticoids) signals negatively impacts cutaneous host defense to pathogens and worsens inflammatory skin conditions such as psoriasis. Toll-like receptors (TLRs) on macrophages and other innate immune cells sense danger signals and trigger innate and adaptive immunity, but whether these stress stimuli affect similar or exclusive TLR-dependent immune pathways is unclear. We hypothesize that quantitative analysis by RNAseq will provide critical information into the mechanism by which stress affects macrophage inflammatory signaling. Using RNASeq, we find distinct regulatory profiles for β-adrenergic agonist and GC on TLR4-mediated inflammatory responses. β-adrenergic stimuli are highly selective inhibitors of a small subset of TLR4-inducible genes, inhibiting less than 15 genes by 5-fold or more (80% inhibition), showing striking selectivity. Three of these genes, Il12b, Il27, and Il6, are induced with similar kinetics and high magnitude, suggesting similar regulation of these genes. In contrast, the glucocorticoid Dexamethasone (Dex) inhibited 22 of the 283 induced genes. Importantly, Dex targeted a different subset of inflammatory genes, including Il1b, Il1a, and Ptgs2. In conclusion, we found that adrenergic and neuroendocrine stress signaling targets distinct macrophage inflammatory responses, which may cooperate to inhibit different immune pathways during a physiologic stress response.
BCAP promotion of TLR-induced IFNα production in plasmacytoid DC (INM7P.346)
Authors: Chu Talyn 
Published: January 2023

Abstract
B cell adaptor for PI3-kinase (BCAP) is a cytosolic adaptor protein involved in phosphatidylinositol-3 kinase (PI3K) activation in several immune lineages. In macrophages, we have shown that BCAP links TLR to phosphatidylinositol-3 kinase (PI3K) activation, thereby negatively regulating the production of proinflammatory cytokine production. BCAP is also expressed in plasmacytoid DC (pDC), in which PI3K signaling has been implicated in TLR responses. Therefore we investigated the role of BCAP in TLR7 and TLR9 responses in pDC. We confirmed high expression of BCAP in pDC by intracellular staining. We then examined pDC responses to TLR7 and TLR9 agonists. pDC grown from bone marrow in Flt3L from BCAP-/- mice secreted lower amounts of IFNa than wild type (WT) pDC in response to the TLR9 agonist CpG-C or TLR7 agonists R848 and ssRNA40. BCAP-/- pDC also had decreased Akt phosphorylation upon TLR7 and TLR9 stimulation, consistent with the function of BCAP to activate the PI3K pathway downstream of TLR in macrophages. BCAP-/- pDC secreted similar or higher levels of IL-6 than WT pDC, similar to inhibition of PI3K in human pDC. We found that BCAP-/- pDC directly isolated from bone marrow also showed reduced production of IFNa in response to CpG-C compared to WT pDC. Therefore, BCAP is a protein that specifically promotes TLR7/9-induced PI3K activation and IFNα production in pDC.