bluuebunny/crossref_metadata_2025_split huggingface dataset:
Creatine shapes macrophage polarization by reprogramming L-arginine metabolism
Authors: Ji Liangliang , Zhao Xinbin , Zhang Bin , Kang Lan , Song Wenxin , Zhao Baohong , Xie Wei , Chen Ligong , Hu Xiaoyu 
Published: January 2023

Abstract
Macrophage polarization is accompanied by drastic changes of L-arginine metabolism. Two L-arginine catalytic enzymes, iNOS and arginase 1, are well-characterized hallmark molecules of M(IFN-γ) and M(IL-4) polarization, respectively. The third metabolic fate of L-arginine is generation of creatine that acts as a key source of cellular energy reserve, whereas little is known about regulation and function of creatine metabolism in the immune system. Here, combination of genetic, genomic, metabolic, and immunological analyses revealed that creatine fundamentally reprogrammed macrophage polarization by suppressing M(IFN-γ) yet promoting M(IL-4) effector functions. Mechanistically, creatine inhibited IFN-γ-JAK-STAT1-mediated induction of immune effector molecules including iNOS while supported IL-4-triggered STAT6-dependent upregulation of arginase 1. Depletion of intracellular creatine by ablation of creatine transporter Slc6a8 significantly altered macrophage-mediated immune responses in vivo. These results depicted a previously uncharacterized regulatory loop whereby creatine, a single cellular metabolite, shaped macrophage polarization by regulating L-arginine metabolic enzymes.
The role of the HO-1/Heme axis on macrophage cytokine production
Authors: Choa Ruth , Kambayashi Taku 
Published: January 2023

Abstract
Transfusion Related Immunomodulation (TRIM) is a phenomenon that was first described over 30 years ago when it was reported that patients receiving blood transfusions prior to renal transplants had improved graft survival rates. The prevailing hypothesis for these observations at the time was that donor leukocytes were responsible for the effects; however, transfusion of leukoreduced and irradiated blood products still resulted in TRIM, suggesting that another component of the blood products, such as red blood cells (RBCs), might mediate the immunosuppressive effects.
RBCs have been shown to affect the activation of immune cells and immune responses. For example, monocytes co-cultured with RBCs and stimulated with LPS produce lower levels of TNF-a than those that are not co-cultured. This suggests that RBCs can shape the immune response that develops during antigen presentation.
Splenic red pulp macrophages are critical for clearance of both RBCs and heme, and these processes are essential for maintaining RBC homeostasis and preventing heme-mediated toxicity. Inside macrophages, heme is degraded by the enzyme Heme Oxygenase-1 (HO-1) and HO-1 activity leads to increased IL-10 and decreased TNF-a secretion. We hypothesized that erythrophagocytosis by macrophages during RBC transfusion may result in the activation of a heme-mediated HO-1 dependent signaling pathway that causes immunosuppressive effects. We used the robust TLR stimulation models of LPS and CpG challenge, and tested whether RBC transfusion or heme signaling could modulate the resulting macrophage cytokine response. We found that CORM-2 (a synthetic CO releasing molecule) downregulates the macrophage pro-inflammatory cytokine profile.
TRAF3 is a critical regulator of the expansion of myeloid-derived suppressor cells in mice
Authors: Zhu Sining , Lalani Almin , Sant’Angelo Derek , Covey Lori R , Xie Ping 
Published: January 2023

Abstract
TRAF3 is an adaptor protein and E3 ligase that regulates the signal transduction pathways of the TNFR superfamily, pattern recognition receptors, and cytokine receptors. We previously showed that specific deletion of TRAF3 in myeloid cells leads to spontaneous inflammation and tumor development in aging M-TRAF3−/− mice, which is consistently associated with a striking expansion of CD11b+Gr1+ myeloid cells. Here we report that these CD11b+Gr1+ cells were myeloid-derived suppressor cells (MDSCs), as they exhibited a strong suppressive activity on CD8 T cell proliferation and also displayed a nuclear morphology of immature myeloid cells. Using a chronic inflammation model induced by repeated injections of heat-killed Bacillus Calmette-Guérin (BCG), we showed that young adult M-TRAF3−/− mice also exhibited hyperexpansion of MDSCs after BCG injections. Our in vivo BrdU labeling experiments revealed that MDSC hyperexpansion in M-TRAF3−/− mice was mainly due to the increased proliferation of MDSCs or progenitor cells in the spleen but not in the bone marrow (BM). Interestingly, TRAF3 does not appear to be an intrinsic regulator of the development, survival or proliferation of MDSCs, as suggested by the results of cultured BM progenitor cells and purified splenic MDSCs, in the absence or presence of CSFs or agonists of TLRs or CD40. Furthermore, we identified environmental factors that induced the aberrant expansion of MDSCs in M-TRAF3−/− mice using cytokine protein array analyses, which were further verified by macrophage cultures and in vivo neutralization studies. Taken together, our findings identify TRAF3 as a new regulator of MDSC expansion, and therefore also a critical regulator of chronic inflammation and immunosuppression.
Distinct heat shock protein-mediated immune responses are induced by macrophages and dendritic cells
Authors: Sedlacek Abigail L , Kinner-Bibeau Lauren B , Wang Yifei , Mizes Alicia P , Binder Robert J 
Published: January 2023

Abstract
Heat shock proteins (HSPs) induce potent immune responses against their chaperoned antigens and thus have potential as cancer immunotherapy. APCs, T cells, and NK cells are all required in HSP-mediated immune responses. NK cells are activated indirectly via APCs, but the mechanism of this activation is poorly understood. Interestingly, MΦ are capable of inducing IFNγ production by NK cells, however, DC are not. This is observed in response to multiple HSPs including gp96, calreticulin and HSP90. Thus, APC production of potential mediators of NK cell activation were investigated. Gp96 induces CXCL10 production in MΦ but not DC and this chemokine is necessary for NK cell activation. Since CXCL10 is STAT1 induced, STAT1 phosphorylation (STAT1p) was examined. A unique pattern of STAT1p is observed in MΦ where S727 is phosphorylated early, followed by delayed phosphorylation of Y701. In contrast, DC exhibit delayed phosphorylation of S727, and phosphorylation of Y701 is not observed. Upstream, p38 MAPK is phosphorylated to comparable levels in both APCs. However, basal expression of the phosphatase SHP2 is higher in DC. Blocking SHP2 in DC leads to earlier phosphorylation of S727 but no recovery of Y701 phosphorylation or subsequent induction of IFNγ production by NK cells. Thus, additional phosphatases and inhibitors are likely contributing to these signaling events. Collectively, these data demonstrate that targeting both MΦ and DC in vivo is critical for optimal HSP-mediated immune responses.
TRIM14 promotes endothelial activation via NF-κB signaling pathway
Authors: Li Yong , Fan Daping , Fu Mingui 
Published: January 2023

Abstract
Endothelial activation induced by proinflammatory cytokines is closely associated to the pathogenesis of atherosclerosis and other vascular diseases, however, the molecular mechanisms controlling endothelial activation are not fully understood. Here we report that TRIM14 promotes endothelial activation by enhancing cytokine-induced NF-κB activation. TRIM14 is highly expressed in human vascular endothelial cells, and markedly induced by TNF-α, IL-1β and LPS in both HUVECs and aortas of LPS-injected mice. Overexpression of TRIM14 significantly increased TNF-induced expression of adhesion molecules such as VCAM-1, ICAM-1, E-Selectin and cytokines in human endothelial cells, and by which it facilitated monocyte adhesion to the activated endothelial cells. Conversely, its silencing has opposite effect on endothelial activation. We further found that TRIM14 interacted with TAK1 and NEMO to facilitate the IKK complex formation and enhanced cytokine-induced phosphorylation of IκBα and NF-κB nuclear translocation. Moreover, we found that TRIM14 underwent K63-linked ubiquitination during TNF-induced endothelial activation, which is required for its positive role in the regulation of NF-κB signaling and endothelial inflammation. Meanwhile, NF-κB can directly bind to the promoter regions of TRIM14 gene and control its mRNA transcription. Finally, TRIM14 protein levels were significantly up-regulated in mouse and human atheroma compared to normal arteries. Taken together, the current study demonstrates the new role of TRIM14 in the regulation of endothelial activation and suggests that TRIM14 may be a potential therapeutic target for vascular inflammatory diseases such as atherosclerosis.
CFTR, which not only serves as a TCR signaling molecule but also function as an anion channel, dual-negatively regulates IFN-γ production and tumor immunity in γδ T cells
Authors: Duan Yuanyuan , Li Guangqiang , Xu Miaomiao , Yin Zhinan 
Published: January 2023

Abstract
γδ T cells have been demonstrated to have the MHC independent-broad anti-tumor activities with high IFN-γ production in the infiltration of the tumor tissue, therefore, they become the strongest interest for cancer immunotherapy. Ion channels play critical roles in the cell functions, especially in the regulation of immune cells functions. Cystic fibrosis transmembrane conductance regulator (CFTR) is an important chloride channel and regulator in the apical membrane of epithelial cells. Recently, CFTR was reported to participate in the immune regulation and likely account for the risk of developing cancer. However, little is known about the effect of CFTR on γδ T cells tumor immunity. In this study, we would like to elucidate the regulatory mechanisms and indispensable functions of CFTR on γδ T cells in tumor immunity. We found that genetic deletion of CFTR displayed the increased TCR signaling and elevates IFN-γ release in peripheral γδ T cells, but not in CD4+ T cells. The absence of CFTR in γδ T cells strengthened the TCR signaling. The underlying mechanism of higher TCR signaling was likely that CFTR may participate in TCR signaling via interacting with the PLCγ-1-LCK-ZAP70 signaling complex in γδ T cells. Consistent with the data in mice, CFTR inhibition, which increased intracellular Ca2+ concentration, significantly increased the IFN-γ production in γδ T cells. Finally, CFTR−/−γδ T cells displayed higher cytolytic ability against the B16 melanoma and protected against B16 tumor growth. This study systematically defined the roles of CFTR in the activation of γδ T cells and its potential contribution to tumor immunity, which will enrich the γδ T cells biology and expand the sight for the application of CFTR in γδ T cells cancer immunotherapy.
PD-1 is negatively regulated by LSD1 through interactions with Blimp-1 during acute viral infection
Authors: Neeld Dennis Kyle , Bally Alexander , Lu Peiyuan , Majumder Parimal , Barwick Benjamin , Boss Jeremy M 
Published: January 2023

Abstract
Prolonged exposure to antigen, such as during chronic viral infections and cancer, leads to sustained TCR signaling and ultimately T cell exhaustion, which is in part mediated by up regulation of Programmed cell death 1 (PD-1). Antibody blockade of PD-1 does not work in all instances and can lead to autoimmunity, thus identifying novel tools to manipulate PD-1 are needed. Therefore, we sought to understand the genetic and epigenetic mechanisms that regulate PD-1 during acute and chronic antigen conditions in order to identify additional therapeutic targets to modulate PD-1. We determined the chromatin status at the PD-1 locus and found that activating H3K4me1 and H3K4me2 epigenetic modifications were still present at day 8 during a chronic but not acute LCMV infection. This suggested a failure to remove the active histone marks in chronic infections. Indeed, CD8 T cells lacking lysine-specific histone demethylase 1A (LSD1), a H3K4 demethylase, display significantly higher levels of PD-1 mRNA and protein during acute but not chronic infection. Molecular analysis revealed that Blimp-1 recruits LSD1 to the PD-1 locus to induce silencing during acute infection. Despite Blimp-1 binding to the PD-1 locus during both acute and chronic LCMV infection, it fails to recruit LSD1 during the chronic setting. Additionally, failure to remove H3K4 methylation resulted in less DNA methylation at known PD-1 control regions compared to wild-type cells, consistent with the differential expression of PD-1. Taken together, these results demonstrate that LSD1 downregulates PD-1 by interacting with Blimp-1 during acute, but not chronic LCMV infection, facilitating full epigenetic repression of the locus.
Simultaneous analysis of mRNA and proteins in immune cells using the BD&[trade] Single-Cell Multiplexing Kit and BD&[trade] AbSeq reagents on the BD Rhapsody&[trade] system for high-resolution interrogation of differential immune regulation
Authors: Song Hye-Won , Baracho Gisele V , Bansal Nidhanjali , Taylor Ian , Shum Eleen , Widmann Stephanie , Mortimer Stefanie 
Published: January 2023

Abstract
Obesity can weaken the body’s immune system and trigger chronic inflammation. To understand the root cause of this problem, we used single cell sequencing to examine thousands of immune cells isolated from primary and secondary lymphoid organs as well as adipose tissue and compared a diet-induced obesity mouse model with control mice. Cells were stained with 30 DNA-barcoded antibodies from BD™ AbSeq reagents to enable multiomic analysis, i.e. examining protein alongside mRNA expression in tandem. We also utilized DNA-barcoded universal antibodies from the BD™ Single-Cell Multiplexing Kit, which allowed us to combine 8 samples from different mice and tissue types into a single pooled sample, significantly reducing experimental scale and cost while eliminating potential batch effects. The pooled samples were loaded on the BD Rhapsody™ system to perform cell lysis and individual mRNA and cell barcoding, allowing measurement of ~400 immune-related mRNAs and 30 proteins at the single cell level. We were able to de-multiplex the pooled samples with high specificity after sequencing. The targeted mRNA and AbSeq panel provided robust clustering of immune cell types and showed that genes related to critical immune responses, including inflammation and lymphocyte activation, are differentially regulated in specific immune-cell subsets in obese mouse. Using this multiomic analysis of genes differentially regulated in immune cells from different tissues, we propose a model to explain the immuno-phenotype we observed in obese mouse.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
BD, the BD Logo, and Rhapsody are trademarks of Becton, Dickinson and Company. © 2019 BD and its subsidiaries. All rights reserved.
Single cell transcriptomics resolves activation dynamics and cellular states of human blood and tissue T cells
Authors: Szabo Peter Anthony , Levitin Hanna M , Miron Michelle , Snyder Mark E , Senda Takashi , Yuan Jinzhou , Chen Yim Ling , Bush Erin C , Dogra Pranay , Thapa Puspa , Sims Peter A , Farber Donna L 
Published: January 2023

Abstract
T cells persist as heterogeneous subsets throughout the body and are essential in mounting protective immune responses. In healthy humans, most of our knowledge of T cell activation derives from sampling the peripheral blood and therefore the transcriptional states of tissue T cells, their functional responses to stimulation, and how they relate to T cells in blood have been poorly defined. Here, we profile the activation dynamics of T cells isolated from human lungs (LG), lymph nodes (LN), bone marrow (BM) and blood following TCR-stimulation by single cell RNA-sequencing (scRNA-seq). Analysis of &gt;50,000 individual resting and activated T cells using clustering and new factorization methods reveals lineage-specific gene expression signatures and discrete activation trajectories in all tissues. Between sites, T cells from LG and LN are most distinct, while blood T cells are most similar to those in BM but persist in a more activated basal state. We identify a common transcriptional profile of tissue T cells and detect trace numbers of these cells in the blood. We also define cellular states for resting and activated T cells across tissues, including an interferon-induced state in CD4+ T cells and distinct effector states specific to CD8+ T cells, and uncover new markers of T cell activation that may be central to T cell function. Importantly, we demonstrate that the T cell activation states resolved here serve as a new baseline for defining T cell dysfunction in disease, revealing novel insights into T cell states among tumor-infiltrating lymphocytes from previous studies. Our investigation couples scRNA-seq with new analysis methods to define the activation dynamics of human T cells from disparate anatomical sites on a single cell level.
Engaging with Pacific Communities to support effective dialogue, appropriate outreach and scientific knowledge translation activity
Authors: Sika-Paotonu Dianne , Anitelea Toni , Uiese Tina , Cockburn Alana 
Published: January 2023

Abstract
Indigenous Māori and Pacific population groups remain disproportionately affected by Acute Rheumatic Fever (ARF) and Rheumatic Heart Disease (RHD). ARF is an autoimmune response to untreated throat (and skin) infection by Group A Streptococcal bacteria. Untreated and repeated episodes of ARF or a severe attack, can lead to permanent cardiac damage known as RHD. High rates of ARF and RHD persist in New Zealand and the Pacific Region. To help raise awareness and support knowledge translation efforts that facilitate dissemination of research work and findings of relevance to Pacific populations, appropriate community engagement is needed. The purpose of this work was to host a Pacific community event to support appropriate engagement and dialogue between researchers and Pacific communities in the southern Wellington Region that would facilitate communication and discussion of ARF/RHD related Penicillin research works and findings. A Pacific community event was held on 30th July, 2018 in Porirua that involved Pacific researchers, University Pacific staff a Medical student and various Pacific groups in the area. Scientific information and research focused on Immunological components related to ARF and RHD were presented by a Pacific Biomedical Scientist with the audience participating in a lengthy discussion session that followed. Feedback demonstrated those involved had engaged appropriately and successfully with the Pacific community groups represented with requests made for similar future events to be held. The response to this Pacific community event highlights the importance of cultural awareness being combined with appropriate scientific and knowledge translation communication activity to support effective dialogue.
Development and evaluation of an interactive simulation to teach undergraduate immunology students all stages of monoclonal antibody (mAb) production
Authors: Costabile Maurizio 
Published: January 2023

Abstract
Monoclonal antibodies (mAbs) have revolutionised science since the method was first reported in 1975. The resultant impact of this technique led to Köhler and Milstein being jointly awarded the Nobel prize in 1984. Given the wide use of mAbs in science, it is important that undergraduate immunology students understand how they are produced including the principles of the selection process. Past experience has demonstrated that some undergraduate Immunology students at the University of South Australia (UniSA) experienced difficulties in learning these principles. While there are numerous resources available to students on the internet, they have their shortcomings, such as the focus being on the selection process alone, omitting the prior steps involved and all these resources are completely passive ways of learning. In an attempt to enhance undergraduate student learning of all aspects of mAb production, an interactive simulation was developed. The impact of the simulation was evaluated in 2017 and 2018 with significant improvements in student performance in a quiz and end of semester examination seen for those students that made use of the simulation. Anonymous student feedback was overwhelmingly positive, with students enjoying the new teaching approach. I suggest that interactive simulations can enhance the teaching of complex immunology topics to undergraduate students. The data clearly shows that this approach has significantly enhanced student learning and now forms a core part of teaching mAb production at UniSA. The simulation is freely available to any teaching staff that wish to include it as part of their teaching approach.
Potassium efflux and serum amyloid A3 are involved in IL-1β production by macrophages infected with Mycobacterium tuberculosis
Authors: Jung Bock-Gie , Samten Buka 
Published: January 2023

Abstract
While interleukin (IL)-1β is essential for the protective immunity against tuberculosis infection, it also exacerbates the pathology of tuberculosis. Although previous studies suggested that early secreted antigenic target of 6-kDa (ESAT-6) of Mycobacterium tuberculosis (Mtb) is a potent activator of NLRP3 inflammasome for IL-1β maturation, the mechanisms remain unclear. Therefore, we studied IL-1β production by C57BL/6 mouse bone marrow-derived macrophages (BMDM) infected with Mtb H37Rv, its esat-6 deletion mutant or esat-6 complemented strain. Our data clearly showed that BMDM infected with esat-6 deletion mutant produced significantly reduced IL-1β compared with the cells infected with H37Rv and esat-6 complemented strain despite having comparable levels of pro-IL-1β, implying that ESAT-6 is critical for the maturation but not synthesis of pro-IL-1β. IL-1β production by Mtb-infected BMDM is inhibited by MCC950, a selective NLRP3 inhibitor. Interestingly blocking K+ efflux by high concentrations of extracellular K+ or quinine, an inhibitor of TWIK2 K+ efflux channel, inhibited IL-1β production, but increased bacilli growth in BMDM. Infection of mice with esat-6 deletion mutant showed defects in lung IL-1β production compared with H37Rv. Recombinant ESAT-6 induced IL-1β production in mouse lung. RNA-seq analysis identified serum amyloid A (Saa) 3 as one of the most highly expressed genes in ESAT-6 inoculated mouse lung. Silencing Saa3 in BMDM led to reduced IL-1β production by H37Rv or ESAT-6. These data together imply that Mtb induces secretion of IL-1β in an ESAT-6 dependent manner with the involvement of NLRP3, Saa3 and K+ efflux channel which is also required for Mtb growth control in macrophages.