bluuebunny/crossref_metadata_2025_split huggingface dataset:
Kamebakaurin inhibits lipopolysaccharide-induced dendritic cell maturation by blocking transforming growth factor-beta activated kinase 1 (116.30)
Authors: Kim Jee Youn , Kim Yeon Jin , Ryu Hwa Sun , Kim Hyung Sook , Roh Eunmiri , Kim Hwan Mook , Kang Jong Soon , Hong Jin Tae , Kim Youngsoo , Han Sang-Bae 
Published: January 2023

Abstract
Transforming growth factor-beta activated kinase-1 (TAK-1) is a serine/threonine kinase in the mitogen-activated protein kinase kinase kinase family. Once activated, TAK-1 will activate p38, JNK, and IKK, followed by activation of transcription factors AP-1 and NF-κB. Due to its role in the regulation of cell survival, differentiation, and inflammation, TAK-1 becomes one of the important drug targets in many human diseases. In this study, we show that TAK-1 can be directly inhibited by natural compound, kamebakaurin. Kamebakaurin inhibited the production of nitric oxide, IL-12, TNF-α, and IL-1β by dendritic cells, but did not affect their maturation process. We also showed that kamebakaurin inhibited the phosphorylation of p38 and JNK, but not ERK, in LPS-treated DCs. Kamebakarin also inhibited the phosphorylation of MEK3/6, upstream kinase of p38, and MKK4/7, upstream kinase of JNK, but not MEK1/2, upstream kinase of ERK. In addition, kamebakaurin inhibited NF-κB pathway, sequentially IKK phosphorylation, IκB-α degradation and nuclear translocation of NF-κB p65. Finally, we proved that kamebakaurin directly blocked autophosphorylation and kinase activity of TAK-1. Taken together, we show that kamebakaurin is a direct inhibitor of TAK-1 and suggest that it might be candidate for the treatment of inflammatory diseases.
Complement alternative pathway is activated in preeclampsia (116.35)
Authors: Zhang Ming , Velickovic Ivan , Dalloul Mudar , Alam Amit , Lai Chih Yuan , Medranda Giorgio , Abulafia Ovadia , Sherer David 
Published: January 2023

Abstract
In a previous study of a cohort of pregnant women (mostly whites, with 4.6% preeclampsia), complement factor B, a central molecule of alternative complement pathway, is observed as a predictor of preeclampsia in early pregnancy. Whether this observation could be applied to other populations is unknown. Further, the status of factor B in the fetal circulation of preeclampsia is unclear. We investigated these important questions by examining a cohort of pregnant women with high risk of preeclampsia. Fifty pregnant women (mostly African Americans, with 14% preeclampsia) were enrolled. Preeclampsia were diagnosed before delivery. Blood samples were collected from maternal and fetal umbilical cord blood immediately after delivery. Plasma Bb levels, the active fragment of factor B, were quantified by ELISA. We found that maternal plasma Bb levels were significantly higher in women with preeclampsia than those of normal pregnancy (increased 72%, P=0.02). In addition, Bb levels in the fetal cord blood of preeclampsia group were also significantly higher than those of normal (increased 92%, P=0.03). In summary, in a cohort of minority pregnant women with high risk of preeclampsia, complement factor B is activated in both maternal and fetal circulations during preeclamptic pregnancy. Activation of alternative complements may contribute to the systemic inflammatory response in both mothers and fetus.
DAP12 negatively regulates toll-like receptor responses in macrophages through SHIP1 and DOK3 (116.5)
Authors: Humphrey Mary Beth , Malhotra Shikha 
Published: January 2023

Abstract
DAP12, an ITAM- adaptor, associates with TREM2 and together they limit cytokine responses to LPS in macrophages. The aim of this study was to identify molecular mechanisms needed for the negative regulation of TLR signals by TREM2 and DAP12. LPS stimulation of macrophages induces the co- immunoprecipitation of DAP12 with src-homology containing inositol phosphatase-1 (SHIP1). LPS induced SHIP1 phosphorylation in wild type but not DAP12- deficient macrophages. LPS stimulated DAP12 or SHIP1-deficient macrophages produced significantly more TNFa than wild type macrophages. Further analysis showed that SHIP1 co-immunoprecipitates with another adaptor protein DOK3 in response to LPS. Phosphorylation of DOK3 was dependent on SHIP1 as SHIP1- deficient macrophages had a defect in DOK3 phosphorylation upon LPS stimulation. Knock down of DOK3 expression in RAW264.7 cells lead to increases in TNF-alpha production in response to LPS. These data indicate that DOK3 has a negative function in regulating LPS response. Thus, our study suggests a novel pathway that negatively regulates TLR4 responses. Upon LPS-stimulation of macrophages, DAP12 recruits SHIP1 which in turn regulates the activation and localization of DOK3. DAP12, SHIP1, and DOK3 are required to control LPS-induced cytokine production possibly via the Erk pathway. Studies are in progress further define the key proteins required for this negative pathway and to validate our findings in mouse models of shock.
EAT-2 links SLAM-family receptors with PLCγ1 to activate human NK cells (116.8)
Authors: Wilson Timothy , Brown Marion 
Published: January 2023

Abstract
Cell surface receptors are important mediators of pathogen recognition and intercellular communication. The SLAM-family receptors are a group of Immunoglobulin Superfamily receptors that signal through the small SH2 domain-containing adapters SAP and EAT-2. These adapters promote cellular activation through interactions with Src-family kinases and PKC-θ, though putative inhibitory signaling can be found under some conditions. Our laboratory previously reported a novel interaction between the adapter protein EAT-2 and PLCγ1, suggesting a potent activating potential for EAT-2 in NK cells. Subsequent experiments have demonstrated that EAT-2 phosphorylated on Tyr-127 specifically recruits PLCγ1 but not PLCγ2. By shRNA-mediated knockdown of human NK cell gene expression, we determined that activation of cytotoxicity through SLAM-family receptors was critically dependent on PLCγ1. Importantly, cytotoxicity through other receptors such as CD2 or NKp30 was unaffected by the absence of PLCγ1. While mouse studies have implicated PLCγ2 as a primary mediator of NK cell activation, our study indicates parallel pathways for induction of calcium flux, and demonstrates the importance of the EAT-2/PLCγ1 pathway in human NK cell activity. Further experiments to measure the individual contributions of SAP and EAT-2 have revealed important roles for these proteins in NK cell cytotoxicity.
Stimulating effect of alternatively spliced Interleukin-4 on cultured human T lymphocytes (117.15)
Authors: Luzina Irina , Lockatell Virginia , Atamas Sergei 
Published: January 2023

Abstract
Interleukin-4 is naturally produced in two splice isoforms, one encoded by exons 1-4 (IL-4), and the other encoded by exons 1, 3, and 4 (IL-4δ2). Our recent studies have suggested that both variants are biologically active in vivo, with partially overlapping, yet distinct effects on immune inflammation. In contrast, experiments in cell culture revealed minimal, if any, independent activity of recombinant IL-4δ2. We hypothesized that the reason for this difference between the in vivo system and cell culture may be the instability of purified recombinant IL-4δ2. To address this hypothesis, we co-cultured primary human T cells with normal human fibroblasts that have been engineered to secrete either IL-4δ2 or IL-4. In this system, IL-4δ2 can act on T cells immediately upon release, and it may be also stabilized by other factors generated by cells. Lymphocytes were labeled with CFSE, and their proliferation was analyzed by flow cytometry after 7 days of co-culture. One to three successive generations of cells were observed. Multiplex ELISA assays revealed a 2.5-fold increase in IFN-γ, a 10-fold increase in TNF-α, and more than a 100-fold increase in IL-1β in the presence of IL-4δ2, compared to control co-cultures or those expressing IL-4. There was no difference in the expression of IL-6 or MCP-1 between the IL-4δ2- and IL-4-expressing co-cultures. Thus, IL-4δ2 is biologically active in vivo and in vitro, and its effects are different from those of IL-4.
Human CD4 and CD8 T cells distinctly upregulate Suppressors of Cytokine Signaling 1 and 3 following Interferon-β treatment. (117.24)
Authors: Schneider Raphael , El-Khoury Lama , Beauseigle Diane , Duquette Pierre , Prat Alexandre , Arbour Nathalie 
Published: January 2023

Abstract
Suppressor of cytokine signaling (SOCS) proteins have been identified as a negative feedback loop to cytokine signaling. Emerging evidence supports the potential role of SOCS in controlling immune disorders, but data regarding their expression in human T cells is still sparse. Interferon-β (IFN-β) is a broadly used and effective treatment for multiple sclerosis (MS), although mechanisms of actions are still incompletely resolved. We postulate that IFN-β mediates beneficial effects through the induction of SOCS in peripheral T cells. We first evaluated the expression of SOCS-1 and SOCS-3 at the mRNA (qRT-PCR) and protein levels (FACS, Western blot, immunocytochemistry) by human CD4 and CD8 T cells following IFN-β treatment. A rapid and significant increase of SOCS-1 and SOCS-3 was observed upon cytokine addition, especially in the CD8 T cell compartment. To mimic patients under IFN-β treatment, both T cell subsets were chronically exposed to physiological doses of IFN-β measured in treated patients. Only SOCS-1 but not SOCS-3 levels were elevated after each IFN-β re-exposure. These effects were more pronounced in CD8 T cells. Our preliminary data indicate different expression of SOCS in untreated and IFN-β-treated MS patients. Our results suggest that SOCS-1 contributes to the IFN-β-mediated beneficial effects in MS and that SOCS proteins are distinctly expressed in human T cell subsets. We are currently testing the functional impact of SOCS-1 on human T cell responses.
MicroRNA-21 targets the vitamin D-dependent antimicrobial pathway in leprosy (147.12)
Authors: Liu Philip , Wheelwright Matthew , Teles Rosane , Komisopoulou Evangelina , Edfeldt Kristina , Ferguson Benjamin , Nehta Manali , Vazirnia Aria , Rea Thomas , Sarno Euzenir , Graeber Thomas , Modlin Robert 
Published: January 2023

Abstract
Leprosy is a human disease in which the clinical manifestations correlate with the host immune response to the pathogen, Mycobacterium leprae. In tuberculoid (T-lep) patients the infection is localized and Th1 cytokines predominate in lesions; whereas, in lepromatous patients (L-lep) the infection is disseminated and Th2 cytokines predominate. Evidence also suggests a role for the vitamin D-dependent antimicrobial pathway in contributing to host defense in leprosy. By profiling miRNAs in leprosy lesions, we identified 13 miRNAs differentially expressed in L-lep vs. T lep lesions. Bioinformatic analysis revealed a significant enrichment of L-lep-specific miRNAs that preferentially target genes important in T-lep lesions, including twelve Th1 and antimicrobial pathway genes. The most differentially expressed miRNA in L-lep lesions, hsa-mir-21, was upregulated in M. leprae-infected monocytes and induced by the mycobacterial virulence factor phenolic glycolipid-I (PGL-I). Hsa-mir-21, by downregulating Toll-like receptor induced CYP27B1 and IL1B and upregulating IL-10, inhibited the vitamin D-dependent antimicrobial peptide gene expression. In contrast, knockdown of hsa-mir-21 in M. leprae-infected monocytes enhanced antimicrobial peptide gene expression. The ability of the pathogen to upregulate hsa-mir-21 which targets multiple genes associated with the immunologically localized disease form provides an effective mechanism to escape the vitamin D antimicrobial pathway.
Endothelial-like cells differentiated from bone marrow-derived mesenchymal stem cells preserve T-cell suppressive function (147.13)
Authors: Kim Jung-Sik , Park Min Jung , Lim Hyun-Ju , Kim Young-Hee , Kim Sang-Joon , Park Chung-Gyu 
Published: January 2023

Abstract
Endothelial-like mesenchymal stem cells (EMSCs) differentiated from human mesenchymal stem cells (MSCs) have great potential in cell therapy for ischemic diseases and in tissue engineering for vascular grafts. Although they was proven to stimulate functional blood vessel formation in vivo, it still need to be investigated whether EMSCs harbor immunomodulatory capacity for applying immune-mediated vascular disorders. In this study, therefore, we compared in vivo angiogenic potential of EMSCs with MSCs and studied their immunosuppressive functions. In the phenotype analysis, differentiated EMSC showed several endothelial cell markers including vWF, TGF-beta and Flt-1. In in vitro angiogenesis analysis by using Matrigel and fibrin, they have augmented angiogeneic potential compared to MSCs. Moreover, EMSCs showed neovascularization potential superior to MSCs in subcutaneous transplantation of fibrin-islet-cell composite. In allogeneic mixed lymphocyte reaction, EMSCs showed preserved immunosuppressive function on T cell comparable to that of MSCs. Apoptosis induction and cell-cycle arrest mediated by cell free-culture supernatant were associated with EMSC-mediated T-cell immunosuppression, as evidenced by the increased P27kip and decreased cdk2, cyclin D2 and Bcl-2. These results suggest that ex-vivo expanded EMSC could be a useful source for postnatal neovascularization and for cell-based therapies of tissue damages involving immune reactions.
Topical application of mesenchymal stem cells effectively reduces corneal surface inflammation by inhibiting Th17 T cells proliferation in mouse chemical burn model (147.16)
Authors: Kim Mee Kum , Han Eui Seok , Jeong Hyun Jeong , Lee Hyun Ju , Jeong So Hee , Wee Won Ryang 
Published: January 2023

Abstract
Mesenchymal stem cells(MSCs) has known to have antiinflammatory properties in various inflammatory disease models, however the in vivo effect of MSCs on the inflammation is variable depending on the administration site for the in vivo delivery. To investigate the mechanism of antiinflammatory effect of topical application of MSCs on chemcal burn model, and to compare whether the effect of systemic application of MSCs is the same as topical introduction of MSCs, we made mouse chemical burn model, delivered MSCs, and evaluated local and systemic changes. It had been conducted in conformance with the FASEB Statement of Principles for the use of animals in research and education. We found that topical application of MSCs reduced corneal inflammation in mouse chemical burn model, however, systemic administration of MSCs could not decrease corneal inflammation in the same model. Direct application of MSCs to inflammatory corneal surface diminished not only infiltration of inflammatory cells in the cornea but also Th17 cells in the draining lymph nodes, on the other hand, systemic administration of MSCs did not. It suggested that systemic application of MSC might not effectively reach the target organ (eye) or the draining lymph nodes, and implied that adequate administration of MSCs might be crucial to maximize efficient antiinflammatory property of MSCs in various inflammatory disesase models.
Modulation of MHC II and costimulatory molecules expression on monocytes by intravenous immunoglobulins (147.17)
Authors: Padet Lauriane , Bazin Renée 
Published: January 2023

Abstract
Intravenous immunoglobulins (IVIg) are therapeutic preparations of human IgG isolated from thousands of plasma donations and are known to have anti-inflammatory effects in many autoimmune diseases. IVIg are also used as prophylactic agent to reduce episodes of allograft rejection but the mechanisms responsible for this effect remain to be determined. In the present work, we showed by using a mixed lymphocyte reaction as an in vitro model of allograft rejection and GvHD, that IVIg strongly inhibited T cell activation. To explain this inhibition, we postulated that IVIg modulated the expression of MHC II and costimulatory molecules expressed on APC and involved in antigen presentation and regulation of T cell activation. To study this hypothesis, PBMC were cultured with or without 10 mg/ml IVIg for 24 hours, followed by flow cytometry analysis of cell surface expression of 8 different molecules. Among these, the expression of 5 was significantly modulated on monocytes. IVIg decreased the expression of MHC II (50%), CD80 (80%), CD86 (52%) and L-ICOS (50%). Interestingly, preliminary results showed a 5-fold increase in the expression of PD-L1, a negative regulator of T cell activation. These results suggest that IVIg impair the presentation of allopeptides to T cells as well as induce negative signalling in these cells, leading to their reduced activation. Altogether these findings could help explain the induction of peripheral tolerance and improvement of graft survival by IVIg.
Osteopontin induced inhibition of dendritic cell polarization towards pDC cells is not exclusively mediated by NKT cells (147.28)
Authors: Habeeb Keith , Durre Kainath , Beal Brooks , Qureshi Mahboob 
Published: January 2023

Abstract
Pediatric post-RSV infection associated bronchial asthma is known to be mediated via an enhanced pulmonary Th2 response. However, adult RSV infection is not known to be associated with this pathology. Further, osteopontin and NKT cell have been reported to be involved with induction of Th1 and Th2 type host-immune responses, respectively. The effect of NKT cells on naïve T helper cells is suggested to be a result of NKT cells’ subsequent modulation of naïve dendritic cells (DCs) towards plasmacytoid DCs (pDCs). So, in our present studies, using an NKT cell KO (CD1d-/-) mouse model, we have studied the role of osteopontin and NKT cells in modulating the polarization of DCs towards pDC in RSV-infected neonatal lungs. Flowcytometry analysis determined that there was an enhanced NKT cell (CD19-CD4+CD1d+) response in the RSV-infected neonatal lungs compared to that in the infected adult lungs. There was a decrease in the NKT cell response to RSV infection in the neonatal lungs upon concomitant administration of exogenous osteopontin as compared to those without osteopontin. Furthermore, in the absence of NKT cells (CD1d-/-), the administration of exogenous osteopontin induced inhibition of DC polarization towards pDC. Thus, differential pDC responses to RSV infection in neonates with exogenous osteopontin administration, even in the absence of NKT cells, warrants investigation into involvement of other factors in the development of RSV-induced bronchial asthma in later life.
Suppression of VαiNKT cell activation by Kupffer cells. (147.5)
Authors: Tyznik Aaron , Ichikawa Shintaro , Cheroutre Hilde , Kronenberg Mitchell 
Published: January 2023

Abstract
Invariant NKT (VαiNKT) cells are a unique subset of T lymphocytes that rapidly produce and secrete large amounts of cytokines following activation. These cytokines have a profound impact on a variety of different immune reactions. However, little is known how VαiNKT cell activation is regulated at steady state or curtailed to avoid the escalation of an out of control inflammatory response. Our preliminary data indicates that Kupffer cells, liver resident macrophages, produce soluble factors that can block the cytokine production of primary VαiNKT cells. Surprisingly, this inhibition acts directly on VαiNKT cells stimulated either by cytokine mediated activation or antigen-specific TCR activation. The Cytokine suppression is unique to Kupffer cells as macrophages from the spleen are able to stimulate VαiNKT cells and have no inhibitory action. These studies have revealed a unique mechanism of VαiNKT cell regulation in the liver.
CD11blo Gr1+ osteoclast precursors are increased in inflammatory arthritis and have myeloid derived suppressor cell function. (148.1)
Authors: Charles Julia , Hsu Lih-Yun , Niemi Erene , Weiss Arthur , Nakamura Mary 
Published: January 2023

Abstract
We have identified a CD11blo Gr1+ bone marrow population of osteoclast precursors (OCP). This population is further defined by the markers Ly6Chi CX3CR1+ CCR2+ CD117- CD11c- and is distinct from previously characterized myeloid precursors and monocyte subsets, suggesting that osteoclasts derive from a distinct precursor analogous to the common dendritic cell precursor. OCP increase in inflammatory arthritis in mice and humans. We find that CD11blo Ly6Chi OCP increase significantly prior to the development of synovitis in zymosan-induced SKG arthritis and remain elevated throughout the course of disease. Like the OCP we identified, monocytic myeloid derived suppressor cells (M-MDSC) are Ly6Chi. We tested our OCP for suppressive activity in vivo using an adoptive transfer model of SKG arthritis. Co-transfer of OCP with SKG CD4+ T-cells into RAG-/- recipients significantly decreased development of inflammatory arthritis compared to transfer of CD4+ T-cells alone, although less dramatically than co-transfer of CD4+ T-cells with wild-type Treg cells. We tested for in vitro suppression of T-cell proliferation by OCP and found that CD11blo Ly6Chi OCP suppress both CD4+ and CD8+ T-cell proliferation. Suppression by OCP is dependent on IFNγ and NOS2, similar to the mechanism described for M-MDSC. Our findings suggest that a myeloid precursor population with both T-cell suppressive activity and the potential to become osteoclasts is expanded in inflammatory arthritis.
Differential requirement for STAT4 in IL-18R expression by CD4 T cells during EAE (148.11)
Authors: McWilliams Ian , Harrington Laurie 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is an autoimmune disease characterized by demyelination of neurons in the central nervous system. In order to study the mechanisms underlying MS, we use the well-described mouse model, experimental autoimmune encephalomyelitis (EAE), during which, the CD4 T cell subsets, TH1 and TH17, are associated with immunopathology. STAT4 is an important TH1 transcription factor that, when activated by IL-12, results in the production of IFNγ. Deletion of STAT4 protects mice from EAE; in contrast, deletion of IL-12 and IFNγ does not ameliorate disease. This disconnect raises the question about the role of STAT4 in EAE. The IL-18R complex is thought to be regulated by STAT4, and it has been shown that IL-18R-/- mice are not susceptible to EAE indicating the importance of this receptor during disease. Due to the association between STAT4 and IL-18R, we have investigated the regulation of the IL-18R complex in vitro as well as during disease and the requirement of STAT4 for its expression. Interestingly, we find that in vitro maximal IL-18R levels are detected on WT TH1 CD4 T cells and that this expression is dependent on STAT4. However, in vivo, we find the IL-18Rα expression is similar on WT and STAT4-/- CD4 T cells during EAE. Taken together, these data indicate an important role for STAT4 in IL-18R expression. Nevertheless, during autoimmune chronic inflammation, other factors promote IL-18R independent of STAT4 signaling.