bluuebunny/crossref_metadata_2025_split huggingface dataset:
Signaling pathway triggered by sBAFF may be impaired in monocytes of the patients with Sjögren’s Syndrome (137.43)
Authors: Yoshimoto Keiko , Tanaka Maiko , Komatsu Sayaka , Setoyama Yumiko , Suzuki Katsuya , Tsuzaka Kensei , Abe Tohru 
Published: January 2023

Abstract
Background and Purpose: B cell activating factor belonging to the TNF superfamily (BAFF) is indispensable for proliferation, differentiation and survival of B cells. Several lines of evidence suggest that elevated production of BAFF seemed to be involved in the development of autoimmune diseases such as Sjögren's syndrome (SS). In our previous study, we investigated that BAFF and IL-6 were abnormally produced in SS monocytes upon stimulation with IFN-gamma. In the present study, we investigated a regulatory mechanism of the production of these cytokines in SS monocytes.
Materials and Methods: Peripheral monocytes prepared from SS patients and normal individuals were stimulated with IFN-gamma or soluble BAFF (sBAFF). The expression levels of sBAFF, IL-6 and transcription factors were measured by ELISA and/or semi-quantitative RT-PCR.
Results and Discussion: IFN-gamma remarkably induced the production of sBAFF and IL-6 in SS monocytes. The production of IL-6 was significantly suppressed by an anti-BAFF antibody. In addition, stimulation of SS monocytes with sBAFF induced robust increase in the production of IL-6. These data suggest that the production of IL-6 in monocytes is induced partly through a signal transduction pathway triggered by sBAFF. We found that the expression levels of several transcription factors were aberrantly elevated in SS monocytes as compared with normal monocytes. These data strongly suggest that sBAFF produced in monocytes acts in an autocrine or paracrine fashion to activate the expression of the IL-6 gene, and that signal transduction triggered by sBAFF may be impaired in SS monocytes. This aberration may underlie the development of SS.
Characterization of CNS antibody-secreting cells, the blood-brain barrier, and autoantibody in the Theiler's model of MS (137.7)
Authors: Li Libin , Brady Jennifer , Lagunoff David , Pachner Andew 
Published: January 2023

Abstract
The determination of the pathogenic role of autoantibody (AuAb) is a challenge in any autoimmune disease, such as multiple sclerosis (MS). This project used Theiler's virus-induced disease to characterize the AuAbs produced in the central nervous system (CNS) by antibody-secreting cells (ASCs).
Mice infected with Theiler's murine encephalitis virus (TMEV) develop an immune-mediated demyelinating disease (TIDD), characterized by progressive weakness, CNS inflammation and demyelination, which represents an excellent model of MS. Flow Cytometry (FACS), ELISpot, and real-time RT-PCR, were used to characterize ASCs and their antibody production in the CNS, and ELISA and immunohistochemistry (IHC) were used to identify autoantibodies and their deposition. We found that ASCs in the CNS were CD138 positive, readily visualized as IgG+ by IHC, and commonly found in clusters of cells in the CNS. AuAbs to CNS antigens were present in the sera, and antibody was deposited in the spinal cord of TIDD mice. The blood-brain barrier, measured by CSF albumin index and Evans Blue injection, was intact.
This study analyzes CNS ASCs and autoantibodies with specificity toward CNS antigens in TIDD, and raises the possibility that AuAb may significantly contribute to disability in this model.
This work was supported by the New Jersey Commission on Spinal Cord Research.
Kinetics of myelin antigen presentation in the CNS during EAE (137.8)
Authors: Murphey Cathi , Forsthuber Thomas G. 
Published: January 2023

Abstract
Despite the plethora of information on immune mechanisms derived from studies of autoimmune experimental encephalomyelitis (EAE), the animal model of multiple sclerosis, little information is available in terms of the kinetics of presentation of myelin antigens by APCs in the CNS. The purpose of this study was to show at which time points and by which APCs myelin antigen (MBP, MOG, PLP) was acquired and presented after induction of EAE. To address this issue, mice were immunized with MOG 35-55 peptide and injected with pertussis toxin on day 0 and 2 and followed for disease. Representative mice were sacrificed every day over forty days and brain tissue was recovered for immunofluorescence analysis. Immunofluorescence and confocal microscopy was used to determine expression of APC cell surface markers and myelin antigens. The results show that T cells or dendritic cells (DC) were rare (&lt;1 cell per 500 fields) in non-immunized mice. In contrast, numbers of CD4+ T cells in immunized mice began to increase at day 3, and numbers of DCs showed a notable increase around day 9. Myelin laden DCs appeared between day 11 and 13. Inflammatory infiltrates peaked between days 15 to day 20, coinciding with peak EAE scores. In conclusion, our data suggest that infiltration of CD4+ T cells precedes substantial uptake and presentation of myelin antigens by CNS APCs, which could have important treatment implications.
Selective expansion of autoreactive T cell clonotypes drives autoimmune diabetes (137.9)
Authors: Li Li , He Qiuming , Wang Bo , Tisch Roland 
Published: January 2023

Abstract
It is well established that T cells are the primary mediators of β cell destruction in Type 1 diabetes (T1D). However, the relative role for different β cell-specific T cell clonotypes in the progression of the diabetogenic response, and the molecular basis for their expansion remain poorly understood. CD4+ T cells expressing the BDC2.5 clonotypic T cell receptor (TCR) and/or which recognize the mimetic BDC (mBDC) peptide are thought to have a key albeit undefined role in β cell autoimmunity of the nonobese diabetic (NOD) mouse. Accordingly, using sIAg7-mBDC multimers, BDC2.5 clonotypic CD4+ T cells were tracked and/or isolated in a temporal manner and analyzed at the single cell level. sIAg7-mBDC-binding (g7-mBDC+) CD4+ T cells were detected in peripheral blood lymphocytes (PBL) and the islets at the onset of β cell autoimmunity in NOD female mice, with the frequency and/or number of g7-mBDC+ T cells increasing with age. A comparison of TCR gene usage by single, g7-mBDC+ T cells isolated from 16 week-old NOD female mice demonstrated dominate usage of TRBV15 and monoclonality based on variable β (Vβ) gene complementary determinant region 3 (CDR3) sequences of BDC2.5 clonotypic CD4+ T cells in the islets but not PBL. Furthermore, the analysis of TRBV15+ g7-mBDC+ BDC T cells revealed the biased TRAV13 Vα gene usage. Furthermore, diabetes could be transferred into NOD.scid recipients by g7-mBDC+ T cells purified from the islets but not from PBL. These data demonstrate that g7-mBDC+ T cells are an early indicator and one of the main mediator of the development of destructive insulitis, and that both clonotypic expansion and preferential usage of TCR characterize islet infiltrating g7-mBDC+ T cells.
Functionality of the Bruton’s tyrosine kinase from Mexican patients with XLA. (138.18)
Authors: Vargas-Hernández Alexander , López-Herrera Gabriela , Rojo-Domínguez Arturo , Mogica-Martinez Dolores , Espinosa-Rosales Francisco J. 
Published: January 2023

Abstract
X-linked agammaglobulinemia (XLA) is a primary immunodeficiency originated from mutations in the gene encoding Bruton's tyrosine Kinase (Btk). Btk is a vital protein in the B cell signaling machinery. L111P and E605G are two recently characterized mutations in Mexican patients suffering XLA; they gave us the basis for a more detailed study of the effects of these mutations in the conformation and functionality of Btk. In silico assays showed that L111 and E605 residues are fundamental for the activations and functionality of the Btk. Btk from the two patients and a healthy control were expressed in the Btk-deficient DT40 cell line, in order to confirm the predictions obtained in silico. The absence of the Btk catalytic activity of the patients was reflected in the lack of the Y551 phosphorylation. This may resulted in a reduced activation of PLC-ã2 that was reflected by reduction of intracellular calcium flux. This work show that L111 and E605 residues are fundamental in the structural conformation of the Btk and that in silico analysis can be a useful tool for the analysis of Btk mutations found in primary immunodeficiency patients.
Supported by CONACyT, project No. 70062
Osteopontin is required for natural killer cell development (138.19)
Authors: Kim Mi Sun , Piao Zheng-Hao , Jeong Mira , Yoon Suk Ran , Chung Jin Woong , Choi Inpyo 
Published: January 2023

Abstract
Natural killer (NK) cells are bone-marrow-derived lymphocytes, which share a common progenitor with T cells. Although a model of NK development from hematopoietic stem cells (HSC) has previously been proposed, the factors and detailed mechanisms driving the differentiation of NK remain to be clearly elucidated. Here we show that osteopontin(OPN) is an important molecule as microenvironmental factor for NK development. The OPN deficient mice evidenced distinct defects in the NK populations in both BM and spleen. However OPN deficient HSC also effectively induced the development of NK cells as control HSC, while the experiment of coculture system and in vivo transplantation system showed that the development of NK cells significantly was defected when HSC culture with OPN absence stromal cells, and transplanted HSC rarely induced into NK cells in OPN deficient mice. Also, the level of OPN expression is much more in stromal cells than the developing NK cells, confirming that OPN is important factor as microenvironment for NK development. The secretion of OPN was induced stimulation with IL15 in the BM stromal cells. The function of IL15, as major factor in NK development, might occurs through OPN which is one of other ways for NK development. . In addition, OPN-driven NK maturation was reduced in T-bet-deficient HPC, suggesting that T-bet is required for OPN-mediated NK development. Collectively, these results show that paracrine OPN signaling drives NK-lineage commitment, thus ultimately promoting NK cell development.
Distinct Lymphoid and Myeloid Progenitors Seed the Thymus and Give Rise to all Thymic Lineages (138.7)
Authors: Serwold Thomas , Ehrlich Lauren I Richie , Weissman Irving L 
Published: January 2023

Abstract
It is unclear whether myeloid lineages develop within the thymus, or whether their presence relies on trafficking of mature cells from the blood. Recent studies suggested that a multi-potent thymus-homing progenitor gives rise to both lymphoid and myeloid lineages. This conclusion is inconsistent with models of hematopoiesis that include a common lymphoid progenitor (CLP) - a cell that has no myeloid potential but gives rise to all lymphoid populations, including B, T, and NK cells.
We have addressed this issue and find that the earliest cells within thymus (commonly referred to as DN1) are lymphoid committed when transplanted in vivo, but when put into certain in vitro conditions, these cells make macrophages and neutrophils. Similarly, CLP, which in vivo are lymphoid committed, when put into these same in vitro conditions, also make myeloid cells at high clonal efficiency. These results bring into question conclusions about the myeloid potential of thymic progenitors that were determined in vitro.
To address the source of myeloid populations within the thymus, we asked whether they developed from an intermediate that expressed the RAG recombinase using RAG reporter mice (SVEX). We found that 20% of neutrophils, but very few or no macrophages or dendritic cells within the thymus come from a RAG expressing progenitor. This suggests that a fraction of RAG-expressing progenitors retain neutrophil potential. However, neutrophils make up approximately 0.0001% of the thymus, suggesting that this potential is very limited. Furthermore, we show that separate myeloid committed and lymphoid committed progenitors home to the thymus, suggesting that the myeloid and lymphoid cells within the thymus are mostly, if not entirely derived from separate seeding cells.
C/EBPb Mediates the Synergistic Upregulation of Gene Expression by IFNg and TNF in Bone Marrow-Derived Mesenchymal Stem Cells (138.9)
Authors: Xu Guangwu , Zhang Yingyu , Zhang Liying , Roberts Arthur I. , Shi Yufang 
Published: January 2023

Abstract
Mesenchymal stem cells (MSCs) are potent immunoregulatory cells that have shown clinical utility in suppressing immunity. IFN-γ and TNF play critical roles in the process of immune response and have been shown to regulate the function of MSCs. However, there is limited data about synergism of IFN-γ and TNF in inducing gene expression in MSCs. Here, we found that IFNγ and TNF synergistically induce the expression of IL-6, and several other immune-related molecules in MSCs in vitro. Knockdown of C/EBPβ by specific siRNA inhibited the expression of the genes induced by IFN-γ alone or TNF alone; importantly, it markedly inhibited the synergistic gene expression induced by the combination of IFN-γ and TNF in MSCs. In addition, IFN-γ alone or TNF alone induced the expression of C/EBPβ in MSCs; and these two cytokines cooperatively induced C/EBPβ expression. This suggests that C/EBPβ is a key transcription factor employed by IFN-γ and TNF in synergistically regulating gene expression. We further found that C/EBPβ similarly mediated synergistic gene induction by IFNγ combined with IL-1β or LPS, suggesting that synergy between IFNγ and other stimuli share C/EBPβ as a common mechanism. Furthermore, while STAT1 is critical in IFN-γ signaling, we found that STAT1 knockdown in MSCs did not affect C/EBPβ expression or the synergistic induction of IL-6 by IFNγ and TNF. Thus, C/EBPβ is not regulated by STAT1. These results will advance our understanding of cytokine interactions in general and their role in MSC immunobiology, which may allow better clinical application of these cells.
Opposing Roles for Lyn and Fyn Kinases in Mast Cell IgG Receptor Signaling (139.4)
Authors: Falanga Yves , Ryan John 
Published: January 2023

Abstract
Mast cells are key players of the innate immune system. In addition to their role in atopic disease, mast cell involvement is strongly suspected in inflammatory diseases such as multiple sclerosis, rheumatoid arthritis, and atherosclerosis. Once activated, they degranulate and produce cytokines, resulting in the recruitment of other immune cells. It is well-appreciated that mast cells are activated via the high-affinity immunoglobulin E (IgE) receptor FceRI. In this study we focus on mast cell activation via the immunoglobulin G (IgG) receptor FcγRII/RIII and downstream signaling by Fyn and Lyn kinases. Lyn-deficient mast cells displayed increased histamine and leukotriene C4 secretion, but no change in cytokine production, suggesting a restricted and negative regulatory role for Lyn kinase in mast cell IgG signaling. In contrast, we show that Fyn kinase is required for IgG-mediated histamine, LTC4, and cytokine production in mast cells, and has a similar role in basophils and macrophages. These data illustrate the importance of Src family kinases in IgG signaling, a means by which mast cells may be activated in chronic inflammatory diseases. Supported by NIH grants 1R01 AI59638 and U19A1077435.
Dissection of the SHP-1 functions in FcεRI signaling (139.8)
Authors: Mizuno Kazuya , Nakata Kazuko , Yakura Hidetaka 
Published: January 2023

Abstract
Although SHP-1, a cytoplasmic protein tyrosine phosphatase (PTP), is shown to be a negative modulator in hematopoietic cells, its precise molecular mechanisms are not well defined. To clarify how SHP-1 regulates FcεRI-mediated signaling in mast cells, we generated bone marrow-derived mast cells (BMMC) from motheaten (me) that lack SHP-1 expression, viable motheaten (mev) that express phosphatase-deficient form of SHP-1, and control (WT) mice and examined downstream signaling events and final outputs following FcεRI crosslinking. FcεRI-induced tyrosine phosphorylation of SLP-76 and LAT, activation of MAPKs, and cytokine production were increased in me- and mev-derived BMMC, in good accordance with the previous implication of SHP-1 as a negative regulator. Since the activity of Syk that is responsible for the phosphorylation of SLP-76 and LAT was comparable in all BMMCs, both molecules might be substrates of SHP-1 in mast cells. In contrast, me-BMMC showed severely impaired PLCγ phosphorylation, FcεRI-induced calcium mobilization, especially extracellular calcium entry, and degranulation. We could not observe these phenotypes in mev-BMMC, thus suggesting that the expression of SHP-1, irrespective PTP activity, is sufficient for the positive regulation of these pathways. The absence of SHP-1 expression disrupted the association between SLP-76 and PLCγ, which might explain, at least in part, the decrease of calcium response in me-BMMC. Collectively, all these results suggest that SHP-1 regulates FcεRI-induced downstream signaling events both negatively and positively by functioning as a PTP and an adaptor protein contributing to the formation of signaling complex, respectively.
The role of Tyrosine kinase 2 in a murine model of allergic asthma (139.9)
Authors: Uebel Caroline , Karwot Roman , Maxeiner Joachim , Soó-Becker Kinga , Lehr Hans-Anton , Finotto Susetta 
Published: January 2023

Abstract
Tyrosine Kinase 2 is a member of the mammalian Janus kinase (JAK) family of protein tyrosine kinases which consists of three more kinases (JAK1-3). Tyk-2 mutation in humans is associated with a completely defective in signaling in response to IFNα and IL-12, abnormal responses to IL-6, IL-10 and IL-23, susceptibility to viral infection and to mycobacterial infection and Hyper IgE Syndrome.
Microscopic analyses showed that asthmatic Tyk2(-/-) mice suffered from extreme airway eosinophilic inflammation as compared to wild type mice.
We found that Th1 differentiation is severely impaired in Tyk2(-/-) mice as shown by decreased IFNγ production by lung CD4+ T cells. By contrast, an increase in Th2 cytokines could be measured in the supernatants of lung CD4+ T cells isolated from Tyk2(-/-) mice.
Preliminary results on T regulatory cell differentiation revealed an ascent of CD4+CD25+Foxp-3+ T regulatory cells in asthmatic Tyk2(-/-) mice by FACS analysis. Consistent with this finding, quantitative Real-Time PCR also showed an increase of the T regulatory cell-specific transcription factor Foxp3 in Tyk2(-/-) mice. As both the T regulatory inducing IL-10-pathway and the T reg inhibiting IL-6-pathway are blocked in Tyk2 deficient mice only TGFβ might be regulating Foxp-3 expression in T regulatory cells in these mice. We next wish to investigate the suppressive function of T regulatory cells in the absence of Tyk-2 in allergic models of asthma.
This work is supported by Graduiertenkolleg 1043 "Antigen-specific Immunotherapy".
Non-depleting anti-CD4 antibody alleviates ovalbumin-induced airway inflammation and T-helper type 2 responses in mice (140.13)
Authors: Wu Cheng-Jang , Lin Chun-Yen , Kuo Ming-Ling 
Published: January 2023

Abstract
Bronchial asthma is an emerged chronic inflammatory disorder of the airways. It is becoming urgent to identify target molecules for the preventive and therapeutic purposes. Non-depleting anti-CD4 monoclonal antibody, YTS177, has been used to induce immunotolerance in allograft transplantation through antigen-specific Treg cells. This study aims for determining whether YTS177 is able to inhibit the ovalbumin (OVA)-induced allergic inflammation in an asthmatic murine model. The injection of YTS177 before and after OVA-sensitization was able to suppress airway hyperresponsiveness, pulmonary eosinophilia, OVA-specific immunoglobulin (Ig) E synthesis, and the production of T helper (Th) 2 cytokines from OVA-activated splenocytes. Furthermore, YTS177 had the therapeutic capacity to relieve the attack of asthmatic symptoms when it was administrated after the mice were sensitized and challenged with OVA. However, the depletion of CD25+ cells did not reverse YTS177-induced hyporesponsive status. The state might be resulted from the angery of CD4+ T cells with the increased level of anergy factor-Cbl-b in activated CD4+ cells cultured with YTS177. Moreover, the high level of cell cycle inhibitor p27Kip1 and increased number of G1 phase arrested-T cells were detected. Therefore, p27Kip1 might be involved in the preventive and therapeutic efficacy of YTS177 for allergic diseases.
Identification of TSLP-dependent mediators in a calcipotriol (Dovonex®) driven mouse model of skin inflammation (140.16)
Authors: Jessup Heidi K , Comeau Michael R 
Published: January 2023

Abstract
The epithelial-derived cytokine thymic stromal lymphopoietin (TSLP) is sufficient to induce asthma or atopic dermatitis (AD)-like phenotypes when selectively overexpressed in transgenic mice. Although T and B cells have been reported to be dispensable for the TSLP-induced inflammation in these models, little is known about the downstream pathways or mediators involved in the inflammatory response driven by TSLP. Topical treatment with vitamin D3 (1α,25-(OH)2D3) or MC903, the low-calcemic analog of vitamin D3, has been shown to induce TSLP expression in epidermal keratinocytes. This induction resulted in an AD-like phenotype similar to that seen in TSLP transgenic mice. 0.005% calcipotriol (Dovonex®) is a synthetic, low-calcemic vitamin D3 analog which is FDA-approved for the treatment of psoriasis. Following topical application of Dovonex® onto mouse ears, TSLP mRNA expression increased over time. Concomitant increases in CCL17/TARC, CCL22/MDC, and IL-6 mRNA were also observed. These increases were dependent on TSLP as an antibody that blocked TSLP signaling abrogated the response. Similarly, a variety of TSLP-dependent protein analytes were increased in the skin following application of Dovonex®. Together these data identify TSLP-dependent mediators that may play a role in the development of skin inflammatory diseases such as atopic dermatitis.
A Novel Anti-Inflammatory Role for Andrographolide in Asthma via Inhibition of The Nuclear Factor-kappaB Pathway (140.17)
Authors: Wong WS Fred , Bao Zhang , Guan Shou Ping , Cheng Chang , Kemeny D Michael , Leung Bernard P 
Published: January 2023

Abstract
Persistent activation of nuclear factor (NF)-κB has been associated with the development of asthma. Andrographolide, the principal active component of a medicinal plant Andrographis paniculata, has been shown to inhibit NF-κB activity. We hypothesized that andrographolide may attenuate allergic asthma via inhibition of the NF-κB signaling pathway. BALB/c mice sensitized and challenged with OVA developed airway inflammation. Andrographolide inhibited OVA-induced increases in total cell count, eosinophil count, and IL-4, IL-5 and IL-13 levels in bronchoalveolar lavage fluid, and reduced serum level of OVA-specific IgE. It attenuated OVA-induced lung tissue eosinophilia and airway mucus production, mRNA expression of E-selectin, chitinases, Muc5ac and inducible nitric oxide synthase in lung tissues, and airway hyperresponsiveness to methacholine. In human lung epithelial cells, andrographolide blocked TNF-α-induced phosphorylation of inhibitory κB (IκB) kinase-β (IKKβ), and downstream IκBα degradation, p65 subunit of NF-κB phosphorylation, and p65 nuclear translocation and DNA-binding activity. Our findings implicate a potential therapeutic value of andrographolide in the treatment of asthma and it may act by inhibiting NF-κB pathway at the level of IKKβ activation. (This work was supported by a BioMedical Research Council grant BMRC06/1/21/19/443)