bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differential expression of IL-17A and IL-17F is coupled to TCR signaling via Itk-mediated regulation of NFATc1 (139.4)
Authors: Gomez-Rodriguez Julio , Sahu Nisebita , Handon Robin , Sacta Maria , Anderson Stacie , Kirby Martha , August Avery , Schwartzberg Pamela 
Published: January 2023

Abstract
Th17 CD4+ effector T cells play important roles in autoimmunity and responses to bacterial infections. The roles of cytokines in regulating the transcription factors STAT3 and RORγT in Th17 differentiation have been extensively studied. However, the role of T cell receptor (TCR) signaling in Th17 differentiation is less appreciated. To examine this issue, we evaluated IL17 production from T cells lacking Itk, a tyrosine kinase required for full TCR-induced activation of PLC-γ and downstream pathways. We find that Itk-/- CD4+T cells exhibit specific defects in the expression of IL-17A despite relatively normal expression of RORγT, RORα and the other Th17 cytokines, including the closely related IL-17F. Defects in IL-17A expression and differential regulation of IL-17A and IL-17F were also observed in vivo in Itk-/- mice challenged with an allergic asthma model. Although Itk-deficient cells have slightly depressed phosphorylation of STAT-3 in response to IL-6, expression of constitutively activated STAT-3 failed to rescue their IL-17A production. In contrast, expression of IL-17A could be rescued by pharmacologically-induced Ca2+ influx or by a constitutively active NFATc1. The effects of Itk on transcriptional complexes and chromatin at the IL-17 locus are under further investigation. Our results suggest that Itk specifically couples TCR signaling strength to IL-17A expression through NFATc1 and that TCR signaling pathways differentially influence Th17 cytokine production.
The LFA-1/ICAM-1 bond: A Spatio-Temporal Cue in PMN Recruitment in Acute Inflammation (140.3)
Authors: Dixit Neha , Simon Scott 
Published: January 2023

Abstract
Polymorphonuclear leukocyte (PMN) adhesion to the inflamed endothelium is in part mediated by LFA-1(CD11a/CD18) which recognizes its ligand ICAM-1. It has long been known that ICAM-1 rapidly forms homodimers and this conformation supports stable bonds specifically with high affinity LFA-1 on the PMN in the presence of shear flow. We hypothesize that shear stress on these dimeric pairs of bonds between high affinity LFA-1 and ICAM-1 result in intracellular signaling that serves as a gatekeeper in the transition from PMN arrest to a migratory phenotype. Imaging PMN rolling on an inflammatory mimetic substrate expressing allosteric activating and blocking antibodies to CD18 under shear flow revealed that calcium flux as well as Talin recruitment was mediated primarily through shear stress acting on high affinity CD18 bonds. Immunofluorescence revealed colocalization of cytoplasmic F-actin, Talin, Rap-1, and phospho-Src within regions correlating with high affinity clusters of LFA-1. In summary, these data sets reveal that an integrin mediated outside-in signal is dependent on shear and the affinity of LFA-1 for its ligand ICAM-1 and that a minimum dimerization of high affinity LFA-1/ICAM-1 bonds elicits the assembly of a macromolecular complex that provides a key navigational step in the transition from PMN arrest to a migratory phenotype.
Development of chemotaxis and cell invasion assays in microfluidic flow cells for use with real-time microscopy. (140.9)
Authors: Conant Carolyn , Kendall Stephen , Schwartz Michael , Ionescu-Zanetti Cristian 
Published: January 2023

Abstract
Many immunological processes in health and disease involve migration, homing, invasion and/or transmigration of cells in response to chemoattractants. In vitro analysis of these phenomena typically involves use of a transwell plate insert to create a two environment experiment. This allows quantitation of either migration or invasion, but the inserts can be cumbersome to handle and are normally processed at the endpoint of the experiment. Here, we demonstrated a microfluidic based method to follow chemotaxis and cell invasion in real time. The method enabled acquisition of high content data by microscopy using microfluidic flow cells which are optimized for imaging. Up to 16 conditions were tested simultaneously. The channels were filled with chemokine or matrix in one half of the channel while the other half was maintained with a neutral buffer. Using this technique, we tested differentiated HL-60 cells’ response to fMLP in real time. We also investigated invasion of serum starved HT1080 and MCF-7 cells into FBS containing matrix and found that as predicted only HT1080 cells successfully invaded the matrix. Using these experimental conditions, the method has the potential to yield insights into the early and ongoing processes that occur during homing and invasion of cells within the vasculature.
Reassessment of innate and adaptive immune pathways in fungal-dependent allergic lung disease (141.1)
Authors: Porter Paul , Abramson Stuart , Delclos George , Kheradmand Farrah , Corry David 
Published: January 2023

Abstract
Genes relevant to allergic lung disease were re-evaluated using our fungal inhalation model. We focused on the asthma phenotype as well as fungal clearance in IL-13, IL-5, Stat6, Rag1, CD1, and TSLP deficient mice. For asthma phenotyping, genetic knock-out (or anti-IL-5 treated) mice were intra-nasally inoculated with 400,000 Aspergillus niger conidia every other day for 16 days, and endpoints measured 24 hours after the final dose. The endpoints measured were airway hyper-reactivity (AHR), bronchial lavage fluid differential cell analysis, and lung IL-4 and IFNγ elispot analysis. For fungal clearance, mice were intra-nasally inoculated with 400,000 Aspergillus niger conidia, and lungs removed at various time points to determine the number of viable conidia remaining. Our results confirmed the requirement of IL-13, Stat6, Rag1, and TSLP for AHR induction; the requirement of Stat6, Rag1, IL-13, IL-5 and TSLP for eosinophil recruitment; the requirement of IL-13, Stat6, Rag1, and TSLP for IL-4 producing cells recruitment. CD1 KO mice showed an exaggerated phenotype with increased AHR and IL-4 production. IL-13, Rag1, and IL-5 were required for optimal fungal clearance. Thus, similar to non-infectious asthma models, conventional T cells, the IL-13 signaling pathway and TSLP were important for disease expression induced by viable fungi. In contrast, non-conventional CD1-resrticted NKT cells were found to be potent suppressors of fungal-dependent allergic lung disease.
Notch ligand Delta-like 4 regulates development and pathogenesis of allergic airway responses by modulating IL-2 production and Th2 immunity (141.15)
Authors: Jang Sihyug , Schaller Matthew , Lukacs Nicholas 
Published: January 2023

Abstract
Activation of the conical Notch pathways have been implicated in Th cell differentiation, but the role of specific Notch ligands in Th2 mediated allergic airway responses has not been completely elucidated. In this study, we show that delta-like 4 (Dll4) was up-regulated on dendritic cells in response to cockroach allergen. Blocking Dll4 in vivo at either the afferent or efferent phases of the immune response enhanced allergen-induced pathogenic responses including airway hyperresponsiveness (AHR) and mucus production via increased Th2 cytokines. In vitro assays demonstrated that Dll4 regulates IL-2 in T cells from established Th2 responses as well as during primary stimulation. Interestingly, Dll4 blockade during the afferent, but not the efferent phase, increased IL-2 levels in lung and lymph node of allergic mice. Moreover, Dll4-mediated Notch activation of T cells during primary stimulation in vitro increased apoptosis during the contraction/resting phase of the response, which could be rescued by exogenous IL-2. Consistent with the role for Dll4-mediated IL-2 regulation in overall T cell function, the frequency of IL-4 producing cells were also significantly altered by Dll4 both in vivo and in vitro. These data demonstrate a regulatory role of Dll4 on both Th2 differentiation as well as on Th2 cytokine production in established Th2 responses.
Artemisinin inhibits lipopolysaccharide-induced nitric oxide production by blocking IFN-β production and STAT-1 signaling in macrophages (142.2)
Authors: Kang Jong Soon , Park Ki Hwan , Lee Hyunju , Han Sang-Bae , Lee Kiho , Park Song-Kyu , Kim Hwan Mook 
Published: January 2023

Abstract
Artemisinin is a well-known anti-malarial drug and has been reported to exert anti-inflammatory and anti-tumor effects. In this study, we investigated the effect of artemisinin on lipopolysaccharide (LPS)-induced NO production in macrophages and the molecular mechanisms responsible for its effects. Artemisinin significantly suppressed NO production and inducible nitric oxide synthase (iNOS) mRNA expression in LPS-stimulated macrophages. NF-κB activity, which is known to be important for NO production and iNOS expression, was minimally inhibited only by highest concentration of artemisinin used and MAPKs were not affected by artemisinin treatment. In contrast, LPS-induced activation of STAT-1 was suppressed by artemisinin treatment in a concentration-dependent manner. Further studies demonstrated that the inhibition of STAT-1 activation by artemisinin might be mediated by blocking interferon-β (IFN-β) production. These results suggest that artemisinin suppresses NO production and iNOS expression, at least in part, by blocking IFN-β production and concomitant down-regulation of STAT-1 signaling.
Long lasting elimination of B cells in NZB/W F1 mice can be induced by NZB but not NZW parental CD8 CTL. (143.12)
Authors: Via Charles , Puliaeva Irina , Soloviova Kateryna , Puliaiev Maksym , Puliaiev Roman 
Published: January 2023

Abstract
B cells are a target in current lupus therapy. In the parent-into-F1 (P-&gt;F1) model of induced murine lupus, lupus is prevented by donor anti-host cytotoxic T lymphocytes (CTL) elimination of B cells. To determine the therapeutic feasibility of CTL in a spontaneous lupus model, we transferred NZB or NZW parental splenocytes (60 or 90 x 106 cells) into 8 week old female NZB/W F1 mice. 2 weeks after transfer, NZB-&gt;F1 mice exhibited significant host B cell reduction and upregulation of B cell Fas consistent with in vivo CTL activity, better seen at the higher dose. By contrast, neither dose of NZW donor cells induced elimination of host B cells below control despite significant donor CD4/CD8 T cell engraftment and donor CD4 upregulation of CD44. At 12 weeks after transfer, NZB-&gt;F1 mice exhibited persistent reduction of host B cells significantly below control at both doses whereas B cells were not eliminated in NZW-&gt;F1 mice at either dose. No mice developed proteinuria &gt;2+. Thus, NZW splenocytes fail to induce detectable B cell elimination short or long term despite engraftment and activation of donor T cells, indicative of defective effector CTL function. In contrast, NZB-&gt;F1 mice support the feasibility of CD8 CTL induction and B cell elimination both short and long term. Studies are in progress to determine if lupus-like disease is also attenuated. If so, more narrowly targeted CD8 CTL may be useful in selectively targeting pathogenic B cells in lupus.
PD-1 expression is required in recent thymic emigrants to establish systemic self-tolerance (143.15)
Authors: Anderson Colin , Uwiera Richard , Thangavelu Govindarajan 
Published: January 2023

Abstract
Homeostatic activation of T cells occurs in both natural and artificially induced states of lymphopenia. Control of this process may be particularly important during the period when the first T cells are exported from the thymus, as recent thymic emigrants (RTE) have a heightened sensitivity to homeostatic cytokines. We tested whether PD-1 is required to prevent lymphopenia-induced immunopathology, and assessed its importance in recent thymic emigrants vs. established peripheral T cells. Established T cells express low levels of PD-1 and established PD-1-/- T cells did not cause overt disease in lymphopenic Rag-/- recipients. In contrast, PD-1 was expressed at high levels on RTE, and PD-1-/- RTE produced from hematopoietic stem cells (HSC) generated a rapidly lethal multi-organ inflammatory disease in the Rag-/- recipients. Disease could be blocked by increasing competition through co-transfer of wild type HSC or by using cytokine receptor deficient or neonatal recipients that have reduced lymphoid space for homeostatic activation. Thus, the PD-1 pathway has a broadly important role in self-tolerance, controlling a key checkpoint in the prevention of lymphopenia driven autoimmunity by RTE. Clinical therapies for autoimmune disease employing lymphoablation and hematopoietic stem cell transplantation will need to take into account functional polymorphisms in the PD-1 pathway, if the treatment is to ameliorate rather than exacerbate autoimmunity.
Oligoclonality and innate-like features in the TCR repertoire of type II NKT cells reactive to a β-linked self-glycolipid (143.16)
Authors: Arrenberg Philomena , Halder Ramesh , Dai Yang , Maricic Igor , Kumar Vipin 
Published: January 2023

Abstract
CD1d-restricted type II NKT cells are distinct from the invariant type I NKT cells and are poorly characterized. A major subset recognizes the β-linked self-glycolipid sulfatide, playing a role in autoimmunity and anti-tumor immunity. Here, we analyzed the TCR repertoire of this type II NKT cell subset in mouse, using flowcytometry, TCR gene sequencing and spectratyping analysis of sorted sulfatide/CD1d-tetramer+ cells. For the TCR β-chain, we found predominant usage of the Vβ8.1/Vβ3.1 and Jβ2.7 gene segments with few dominant clonotypes of restricted CDR3 lengths and conserved acidic residues. The TCR α-chain was predominantly encoded by Vα1/Vα3 and Jα7/Jα9 gene segments and showed defined CDR3α regions of 8 or 9 a.a. lengths comprising conserved basic residues. CDR3 regions of both TCR α- and β-chains were encoded by germline sequences or N-additions. Though potential CD1d-binding tyrosine residues in the CDR2β region are conserved between most type I and type II NKT TCRs, CDR1α and 3α regions differ significantly between the two subsets. Collectively, these findings emphasize innate-like features of sulfatide-reactive type II NKT cells as well as oligoclonal TCR Vα-Jα/Vβ-Jβ gene usage characteristic of the antigen-specific responses of conventional MHC-restricted T cells. These data provide important clues to the recognition of β-linked glycolipids by CD1d-restricted T cells in general. (Supported by grants from NIH, JDRF, MSNRC and DNRG (to VK) and DFG (to PA).)
Cross Talk Between Pathogenic CD4+T Cells and B Cells (143.35)
Authors: Xiang Yufei , Peng Jian , Tai Ningwen , Hu Changyun , Zhou Zhiguang , Wen Li 
Published: January 2023

Abstract
The effect of B cell depletion on diabetogenic T cells is not fully understood. We generated hCD20BDC2.5 NOD mice to specifically address this question. We treated hCD20BDC2.5 NOD mice with 2H7 or control IgG.Our results showed that dividing BDC2.5 cells from 2H7 treated mice produced more IFNg than their counterparts from IgG treated mice (P=0.0035). Interestingly, circulating IFNg in serum of IgG treated mice was higher than that in 2H7 treated mice (p=0.0029). To investigate effect of B cell depletion on diabetogenicity of BDC2.5 T cells, we transferred purified CD4+ BDC2.5 T cells, from either 2H7 or IgG treated mice, into NOD/SCID mice. The overall incidence of diabetes was much higher in NOD/SCID mice received BDC2.5 CD4+ cells from 2H7 treated donors than that from IgG treated donors. To investigate the direct effect of B cells on diabetogenic CD4+ T cells, we co-transferred BDC2.5 CD4+ T cells with purified B cells from NOD mice. Interestingly, we found that B cells may have a protective effect as diabetes development was delayed in the NOD/SCID mice received more B cells. In vitro, the addition of B cells inhibited the proliferation of BDC2.5 CD4+ T cells, supporting the in vivo diabetes development results. In summary, B cell depletion affects pathogenic T cells and there is a dichotomy in the role of B cells in diabetes development.
Protection against lupus nephritis in MRL-lpr mice expressing a natural autoantibody transgene (143.39)
Authors: Chen Qing , Mannoor Kaiissar , Matejuk Agata 
Published: January 2023

Abstract
Natural autoantibodies (NAA) are antibodies present in normal sera without exogenous antigen stimulation. They often have relatively low yet specific reactivity towards multiple self and non-self antigens. Despite their potential importance, the functional role of NAA is poorly understood. We recently generated an Ig site-directed transgenic (sd-tg) mouse model carrying a prototypic NAA that binds a variety of antigens including DNA and phosphocholine. Using this model, we have shown previously that the NAA-producing B-cells are positively selected in normal mice. In the present study, we show that expression of NAA in autoimmune MRL-lpr mice results in nearly complete protection from autoimmune nephritis as assessed by proteinuria, kidney histopathology and immune complex deposition. Accordingly, the NAA sd-tg mice have significantly improved survival. The non-tg and tg MRL-lpr mice have very different IgG subclasses: while the non-tg MRL-lpr mice have high titers of IgG2 and IgG3, both of which are highly pathogenic in lupus nephritis, the sd-tg mice have a predominance of IgG1, the least pathogenic IgG subclass. In addition, the tg MRL-lpr mice have fewer CD44hiCD62L+/- activated T-cells and more CTLA-4+ T-cells as compared with the non-tg mice. These results indicate that NAA and NAA-producing B-cells play an important role in protection from lupus nephritis, and suggest that NAA B-cells may have regulatory effects on CD4+ T cells.
A rapid and efficient method for the isolation of distinct human regulatory T cell populations using combinations of antibody mediated buoyant density centrifugation and/or column-free immunomagnetic cell separation (143.41)
Authors: Kokaji Andy , MacDonald Neil , Fairhurst Maureen , Thomas Terry 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are a specialized subset of T cells that play a key role in immune regulation. Harnessing the suppressive function of Tregs is a major area of interest as they hold great potential for the treatment of autoimmune disorders. The first step towards quality Treg research is the isolation of highly purified, functional Tregs. With unique cell isolation platforms, STEMCELL has developed a full range of products for the rapid and efficient isolation of highly functional Tregs from virtually any peripheral blood sample. RosetteSep is an antibody mediated buoyant density centrifugation method used to isolate unlabelled cells specifically from whole blood or buffy coat samples. EasySep is an immunomagnetic cell separation method used to isolate cells from fresh or previously frozen PBMCs. Treg pre-enrichment is achieved by antibody mediated crosslinking of unwanted cells to either red blood cells (RosetteSep) or magnetic particles (EasySep) allowing their removal by Ficoll centrifugation or magnetic separation, respectively. RosetteSep or EasySep pre-enriched Treg populations consist of CD4+, CD4+CD127low or CD4+CD127lowCD49d- T cells. Pre-enriched Tregs can be further purified using EasySep positive selection to isolate Tregs expressing high levels of cell surface CD25. Purities of 85% +/- 10% CD4+CD25highFOXP3+ human Tregs can be achieved depending on the Treg population. From start to finish, Treg isolations can be completed in less than 3 hours.
Sulfatide-mediated activation of type II NKT cells prevents hepatic ischemic reperfusion injury (143.43)
Authors: Arrenberg Philomena , Maricic Igor , Kumar Vipin 
Published: January 2023

Abstract
Hepatic ischemic reperfusion injury (IRI) is a major complication of liver transplantation and resectional hepatic surgeries. Recent studies suggested involvement of NKT cells in the inflammatory response leading to hepatocellular damage. However, no detailed analysis on the role of the distinct type I and type II CD1d-restricted NKT cells has been carried out. We have identified a major subset of type II NKT cells reactive to the self-glycolipid sulfatide. Here, we show that mice lacking type I NKT cells (Jα18-/-) are protected from hepatic IRI, as reflected by reduced hepatocellular necroses and serum ALT levels. In contrast, sulfatide-mediated activation of type II NKT cells is protective, associated with reduced intracellular IFN-γ expression by type I NKT cells. These data are consistent with our earlier report of anergy induction in type I NKT cells following sulfatide administration. Prevention of hepatic IRI also involved inhibition of hepatic recruitment of myeloid cell subsets, especially the CD11b+Gr-1int and Gr-1- subsets, and of NK cells. Our findings clearly indicate a pathological role for type I NKT cells and a protective role for sulfatide-reactive type II NKT cells in hepatic IRI. Our data have important implications for the development of novel strategy for amelioration of clinical problems associated with hepatic reperfusion, including liver transplantation. (Supported by grants from NIH, JDRF, MSNRC and DNRG (to VK) and DFG and YAEL Foundation (to PA)).
Breaking B cell tolerance by multimerized self antigens on follicular dendritic cells (143.7)
Authors: El Shikh Mohey Eldin , El Sayed Rania , Szakal Andras , Tew John 
Published: January 2023

Abstract
Self antigen-bearing follicular dendritic cells (FDCs) appear to be involved in the pathogenesis of autoimmunity, and interfering with the development of FDC-reticula attenuates ectopic germinal centers (GCs) & auto-Ab production. FDCs multimerize monomeric antigens in immune complexes (ICs) and present them polyvalently in an array that extensively cross-links BCRs and induces T cell independent Ig responses with FDC-BAFF & -C4bBP providing co-signals. We hypothesized that self antigens presented as multimerized ICs on FDCs would break B cell tolerance and that would be indicated by induction of autoreactive GCs & auto-Ab responses. To test this, mice were challenged with murine TNF-α or IgE in the form of ICs. Sera were assessed for auto-Ab production, and draining lymph nodes were examined for autoreactive GCs and plasma cells. Murine TNF-α & IgE ICs were retained on well-developed FDC reticula and numerous GCs & plasma cells were induced. Murine TNF-α & IgE-specific IgM auto-Abs were detectable in 48 hrs, and switched to IgG by day 6. This is the first report indicating that B cell tolerance can be broken by presenting self antigens and co-stimulatory molecules on FDCs. The ability to deliberately break B cell tolerance and induce self-regulated auto-Abs provide potential novel therapies targeting endogenous mediators of chronic inflammation & hypersensitivity diseases.