bluuebunny/crossref_metadata_2025_split huggingface dataset:
CARD-only proteins (COPs) inhibit uric acid crystal-induced inflammasome activation and ameliorate gout
Authors: Devi Savita , Indramohan Mohanalaxmi , Jager Elisabeth , Carriere Jessica , Chu Lan H , Almeida Lucia , Greaves David , Stehlik Christian , Dorfleutner Andrea 
Published: August 2023

Abstract
The NLRP3 inflammasome mediates the response to monosodium urate (MSU) crystals and is responsible for the debilitating symptoms of gout. ASC is the essential inflammasome adaptor that bridges NLRP3 to caspase-1 and inflammasome assembly is via sequential homotypic PYRIN domain (PYD)-PYD and caspase recruitment domain (CARD)-CARD interactions between NLRP3 and ASC and ASC and caspase-1, respectively. While controlled inflammasome activation is essential for host defense and wound healing, dysregulated and excessive inflammasome responses cause inflammatory diseases. Hence, proper control is key for a balanced inflammasome response. Humans, but not mice, encode a family of 3 small CARD-only proteins (COPs): CARD16, CARD17 and CARD18, but their role in controlling inflammasome responses in vivo is unknown, which is the focus of our study. COP transgenic mice show reduced MSU crystal induced release of IL-1b and consequently ameliorated gout symptoms. Transgenic macrophages and COP expressing THP-1 cells show impaired MSU crystal-mediated NLRP3 inflammasome and caspase-1 activation, cytokine release and pyroptosis, while COP KO results in a hyper response to MSU. Also, COPs show unique binding pattern to the CARD of caspase-1 and ASC in naïve, primed and MSU-activated macrophages and interfere with the essential CARD-CARD-mediated caspase-1 self interaction and the ASC-caspase-1 interaction by a competitive binding mechanism. We demonstrate that COPs are functional in mice and inhibit NLRP3 inflammasome assembly by a competitive binding mechanism that prevents cytokine release and pyroptotis and ameliorates inflammatory disease. Altogether, our findings illustrate that the COPs have a role in maintaining homeostasis.
National Institutes of Health. (AI099009 and AR064349 to C.S, AI134030, AI140702 and AI120625 to C.S. and A.D.)
TLR-mediated immune activation is a strong suppressor of the Treg-attracting chemokine CCL22
Authors: Gärtig Jan , Grün Juliane , Perleberg Carolin , Schnell Konstantin , Beimert Antonia , Knott Maximilian Martin Ludwig , Röhrle Natascha , Bahner Kevin , Layritz Patrick , Endres Stefan , Anz David , Piseddu Ignazio 
Published: August 2023

Abstract
The immunoregulatory chemokine CCL22 is mainly secreted by dendritic cells (DC) in lymph nodes and plays a crucial role in maintaining the balance between immune tolerance and efficient immune response by induction of intercellular contacts between DC and regulatory T cells (Treg) expressing the only so far known CCL22 receptor CCR4. In the absence of CCL22, the immune response to vaccination and cancer has been proven to be enhanced. In this work, we aimed to understand how activation of Toll-like receptors (TLR) modulates CCL22 levels mechanistically in vitro and in vivo. We observed that a bacterial infection with salmonella typhimurium and treatment with different TLR agonists such as Poly (I:C), LPS, R848 and CpG led to a potent and long-lasting suppression of systemic CCL22 in vivo. Furthermore, using in vitro co-cultures of DC, B cells and T cells, we identified TLR-mediated CCL22 suppression to be mediated by soluble factors derived from non-DC. Additionally, we pursued to investigate the underlying mechanism and could show in vitro that CCL22 suppression is particularly driven by redundant action of IL-10, IFN-α and IFN-γ. Moreover, inhibition of NF-κB reverted the CCL22-suppressing effect of CpG on splenocytes. In CpG-treated mice, Treg/DC contacts were diminished to the level known from Ccl22−/− mice, an observation which links TLR-mediated CCL22 suppression to an immunological relevance. Taken together, we demonstrate that TLR signaling results in CCL22 downregulation via different cytokines, a mechanism which is potentially important to enable the induction of a strong immune response and efficient pathogen clearance in the setting of microbial infection.
Lithium chloride attenuates sepsis-induced acute respiratory distress syndrome via decreasing the expressions of TNF-α, IL-6, CXCL1, and pulmonary edema
Authors: Lin Pei-Yun , Shih Fu-Chia , Yang Chi-Hsuan , Chen Chia-Ling 
Published: August 2023

Abstract
Sepsis is a life-threatening illness caused by an excessive host response to infection. Acute respiratory distress syndrome (ARDS), a serious complication of sepsis, is characterized by pulmonary edema and alveolar barrier injury. However, there is no effective drug proven to reduce the mortality of ARDS yet. Lithium chloride (LiCl) is an inhibitor of glycogen synthase kinase-3β (GSK-3β). Recent studies reveal that GSK-3β inhibition exhibits anti-inflammatory effects in sepsis thereby ameliorating lipopolysaccharide (LPS)-induced acute lung injury. Given that GSK-3β inhibition has been investigated for various diseases, whether LiCl exerting a novel therapeutic effect for treating endotoxemia-induced ARDS remains unclear. Here, we investigated the effects of LiCl on endotoxemia-induced lung inflammation, respiratory failure, and the pulmonary barrier integrity of respiratory membrane. Mice intraperitoneally injected with LPS could induce a significant lymphopenia, lung inflammation with amount immune cells infiltration, and high mobility group box 1 (HMGB1) expression in lung supernatants. LiCl treatment ameliorated lung inflammation and lung function decline in LPS mice. In addition, LPS-induced TNF-α, IL-6, as well as CXCL1 were significantly lessened after LiCl treatment. The measurements of E-cadherin/VE-cadherin expression in lung tissues and IgM in bronchoalveolar lavage fluids indicated that LiCl could partly ameliorate LPS-mediated loss of pulmonary barrier integrity. Accordingly, LiCl effectively attenuated sepsis-caused lung inflammation, lung function decline, and alveolar barrier injury, which suggested that LiCl might act as a potential repurposing drug for treating ARDS in future.
Anti-rheumatic effect of KMU-11361 in various rheumatoid arthritis-related cells targeting multiple protein kinase
Authors: Baek Hye Suk , Hong Victor Sukbong , Lee Hyun-Su , Son Chang-Nam , Kim Sang-Hyon , Jeong Gil-Saeng , Jeon Jieun , Lee Jinho , Kim Shin 
Published: August 2023

Abstract
Protein kinases important for regulating protein phosphorylation, which is a promising therapeutic target for inflammatory disease, such as rheumatoid arthritis (RA). In this study, we synthesized a novel multi-protein kinase inhibitor, KMU-11361, a derivative of indolin-2-one and investigated the mechanisms of anti-inflammatory and anti-rheumatic effects using THP-1 cells, RAW264.7 cells, Jurkat T cells, and human RA fibroblast-like synoviocytes (RA-FLS). Our results revealed that KMU-11361 inhibited LPS-induced upregulation of inducible iNOS, and COX-2, and suppressed LPS-induced phosphorylation of TAK1, JNK, inhibitor of NF-κB kinase α/β (IKKα/β), IκBα, and NF-κB p65 as well as nuclear translocation of NF-κB p65 in THP-1 cells. Furthermore, KMU-11361 attenuated proinflammatory cytokines such as IL-1β, TNF-α, and IL-6, and, remarkably, inhibited LPS-induced activation of the NLRP3 inflammasome in THP-1 cells. Additionally, in activated Jurkat T cells, KMU-11361 inhibited the TAK1-IKKα-mediated NF-κB pathway as well as the phosphorylation of MAPKs. Moreover, KMU-11361 inhibited anti-CD3/CD28 antibodies-induced upregulation of proinflammatory cytokine IL-2. Moreover, it suppressed LPS-induced upregulation of proinflammatory cytokines, p-TAK1, p-IKKα/β, and p-NF-κB p65 in RA-FLS. Surprisingly, KMU-11361 also inhibited the receptor activator of NF-κB-induced osteoclast differentiation and up-regulation of nuclear factor of activated T-cells, NFATc-1 and c-Fos in RAW264.7 cells. Taken together, our results suggest that KMU-11361 has anti-inflammatory and anti-rheumatic effects in various RA-related cells and could be developed as a potent anti-rheumatic drug.
FoxO6 attenuates neuroinflammation by the regulation of anti-inflammatory factors
Authors: Kim Mi Eun , Lee Jun Sik 
Published: August 2023

Abstract
Forkhead transcription factor (FoxO) plays key roles in the induction of various downstream genes involved in the inhibition of cellular metabolism, cell death, cell cycle, and oxidative stress responses. FoxO6 is expressed in the liver, skeletal muscle, and the hippocampus of the brain. FoxO6 also has been proposed to play a role in the promotion of insulin resistance, and inflammation. In this study, we tried to determine the neuroinflammation inhibitory response and signaling mechanism for FoxO6. In order to confirm the inhibitory function of neuroinflammation, FoxO6 knock out cell line and mice were prepared and their functions were confirmed. We have confirmed that neuroinflammation is suppressed in FoxO6 knock out cell line and mice. These findings may suggest that FoxO6 is a novel factor in the regulation of neuroinflammation.
Supported by grants from NRF (NRF-2020R1A2C1012984)
Extracellular vesicles derived miRNAs from bovine milk modulate the inflammatory response to lipopolysaccharide in vitro
Authors: Rosa Fernanda , Santos Rafaela , Lugo Tomas , Brown Audrey , Bickel Ulrich 
Published: August 2023

Abstract
Bovine milk is composed of a variety of biologically active compounds including mi-croRNAs (miRNA), which are found within extracellular vesicles (EVs). Extracellular vesicle-derived miRNAs may play a crucial role in the regulation of the immune response. Thus, this study aimed to identify specific miRNAs present in bovine milk that modulate the immune system’s response to infection. To address this, RAW-Blue™ Cells, a mouse macrophage reporter cell line was challenged with E. coli lipopolysaccharide (LPS) and incubated with a pool of miRNAs isolated from bovine milk through a 24-hour period. After incubation, QUANTI-Blue™ assay was used to detect the activation of the transcription factor NF-κB by the different treatments. Raw-Blue™ cells treated with miRNAs induced to some extend the signaling pathways leading to the activation of NF-κB relative to control cells (P&lt;0.01). Bovine-milk derived miRNAs suppressed LPS-induced NF-κB activity in a dose-dependent manner (P=0.01). To identify the top expressed miRNAs that are driving such effects in vitro, real-time qPCR assays were performed in the milk samples. The miRNA-profiling revealed the presence of miRNAs involved in tumor suppression and cell apoptosis. Overall, our preliminary findings indicated that miRNAs derived from bovine milk have the potential to modulate the immune response to LPS in vitro.
Texas Tech University, School of Veterinary Medicine
Trogocytic-molting of T-cell microvilli controls T-cell clonal expansion
Authors: Jun Chang-Duk , Park Jeong-Su , Kim Jun-Hyeong , Soh Won-Chang , Kim Na-Young , Lee Kyung-Sik , Kim Chang-Hyun , Chung Ik-Joo , Lee Sunjae , Kim Hye-Ran 
Published: August 2023

Abstract
Internalization of the T-cell antigen receptor (TCR) is intimately linked to T-cell activation: a phenomenon thought to be related to the “exhaustion” of T-cell responses. To date, however, no report has considered that during physical interaction with cognate antigen-presenting cells, T cells release many TCRs via T-cell microvilli particles, which are derived from finger-like membrane structures (microvilli) in a combined process of trogocytosis and enzymatic vesiculation and correspond with the loss of membrane TCRs and many external membrane components. Surprisingly, in contrast to TCR internalization, this event leads to rapid upregulation of surface TCRs and remarkable metabolic reprogramming of cholesterol and fatty acids synthesis to meet the demands of clonal expansion, which drives multiple rounds of division and cell survival. We called this event “trogocytic-molting,” which represents an intrinsic molecular basis of T-cell clonal expansion by which T cells gain increased sensitivity to low antigen concentrations.
This work was supported by the Creative Research Initiative Program (2015R1A3A2066253); Bio-Synergy Research Project (2021M3A9C4000991); Bio & Medical Technology Development Program [2020M3A9G3080281] through National Research Foundation (NRF) grants funded by the Ministry of Science and ICT (MSIT), the Basic Science Program (2019R1C1C1009570 & 2022R1A2C4002627) through National Research Foundation (NRF) grants funded by the Ministry of Education (MOE), and supported by Global University Project (GUP), GIST Research Institute (GRI) IBBR grant funded by the GIST (in 2021-2022), and the Joint Research Project of Institutes of Science and Technology (2021–2022), Korea.
3D co-culture models demonstrate radiation-mediated activation of tumor-associated macrophages
Authors: Kaur Aanchal Preet , Baird Jason , Alice Alejandro F , Ball Megan , Kramer Gwen , Crittenden Marka R , Gough Michael J 
Published: August 2023

Abstract
Type I IFN signaling mediated by STING pathway is important for the induction of radiation-stimulated adaptive immune responses in responsive tumors. Macrophages recruited to tumors following radiotherapy (RT) can influence the response to RT. Reprogramming immunosuppressive M2-like macrophages in tumors to exhibit a more activated M1-like phenotype can help improve response to therapies. Here, we hypothesize that a 3D co-culture model of cancer cells and macrophages will be more contextually relevant to the events occurring in tumors and can be used to study the crosstalk that occurs between these cells in vivo. We initially optimized seeding densities of murine MC38 colorectal carcinoma cells to develop spheroids of consistent size. Using GFP+ macrophages and Hoechst stained MC38, we imaged the development and localization of these cells during spheroid formation. Next, co-culture spheroids were exposed to 12 Gy radiation on day 1. Viability and phenotype of macrophages in spheroid co-cultures was assessed on day 4 using flow cytometry by staining for CD80, CD86, MHC Class II, CD69 and CD206 expression. STING ligand, cyclic di-AMP (CDA) treated spheroid co-cultures were used as positive controls. CDA treatment significantly increased CD80/86 expression in 3D co-cultures. Treatment with 12 Gy radiation showed similar effect. Interestingly, macrophages derived from MyD88 Lyz2Cre, MyD88fl/fland STING knockout mice also showed a significant increase in CD80/86 and CD40, suggesting that macrophage activation was independent of MyD88 or STING signaling. Ongoing studies aim to explore the pathways activated on irradiation in these cells and further cross-validate with the macrophage phenotypes observed in vivo in response to RT.
PIEZO1 Deletion in CD11b+ Cells Suppresses Rhabdomyosarcoma Tumor Rejection and Generates a Tumor-Permissive Immune Landscape
Authors: Bonner Melissa Romain , Tomchuck Suzanne , Askew David , Abiff Muta , Huang Alex Y 
Published: August 2023

Abstract
In several cancers, including the aggressive sarcoma, rhabdomyosarcoma (RMS), tumor-associated myeloid cells (TAMCs) are the largest immune populations and are correlated with poor survival outcomes. TAMCs are a mixed and ‘plastic’ population that can sense changes in their environment, including intra-tumoral ECM stiffness, and change their functioning accordingly. Despite the clinical influences these cells have and the detrimental effects that elevated tumor microenvironment (TME) stiffness has on immune cells’ phenotype, little is known about the proteins that sense and mediate these changes. We propose inhibition of mechanosensitive cation channel, PIEZO1, promotes a pro-tumor phenotype in TAMCs while its activation skews this population to anti-tumor functioning thereby enhancing tumor rejection. PIEZO1’s influence on tumor rejection was assessed by inoculating a novel murine CD11b-conditional homozygous PIEZO1 knockout strain with syngeneic RMS cells and performing several analyses. Tumor volume and survival analyses found that RMS-bearing CD11b-conditional PIEZO1 KO mice developed larger tumors and had diminished overall survival as compared to wild-type mice. Flow cytometric analysis of tumors revealed increased pro-tumor macrophage frequency in tumor and expansion of MDSCs in the TME and tumor-draining lymph node (TDLN). Interestingly, despite PIEZO1 KO tumors having increased T cells, they had lower PD-1+ CD4 and CD8 T cell frequencies; a trend also observed in the spleen and TDLN. These findings suggest without PIEZO1 expression, TAMCs predominantly polarize to a ‘pro-tumor’ state, thereby preventing T cell and adaptive immune cells’ activation, leading to lower survival and increased tumor burden.
NCI T32 (5T32CA059366-27)
The ubiquitin ligase Itch skews light zone selection in germinal centers
Authors: Renshaw Lindsay R , Kim Peter , Crici Macaul , Fazelinia Hossein , Spruce Lynn , Oliver Paula M , Moser Emily 
Published: August 2023

Abstract
Immunoglobulin diversification occurs in peripheral lymphoid organs, after the central tolerance checkpoints during B cell development. High levels of somatic hypermutation (SHM) of Ig genes occurs in germinal center (GC) B cells, followed by selection of mutated clones. This process is essential for generation of high affinity antibody to pathogens, but can also produce autoreactive Ig receptors. Selection mechanisms that govern antigen-driven Ig diversification are still incompletely understood. The E3 ubiquitin ligase Itch prevents the emergence of autoimmune disease and autoantibodies in humans and mice, and patients lacking Itch develop debilitating, multi-faceted, potentially fatal, autoimmune disease; yet how Itch regulates GC B cell fate or function is not well understood. By studying Itch deficient mice, we have recently shown that Itch directly limits GC B cell activity to shape antibody responses. Proteomic profiling of GC B cells uncovered that Itch deficient cells exhibit high mTORC1 and Myc activity, hallmarks of positive selection. Mixed bone marrow chimeras revealed that Itch acts within B cells to limit proportions of light zone GC cells, and Itch deficient light zone GC cells were mainly in G1/G0 rather than in cell cycle. These results support a novel role for Itch in skewing selection of GC B cells to restrict LZ accumulation and shape GC-derived humoral immunity. Determining how B cells integrate cues within GCs to navigate through LZs and DZs will aid in understanding how autoreactive clones emerge from GCs in those suffering from autoimmune disease.
Supported by a grant from NIH (K22AI148597)
Deletion of Vβ3+ CD4+ T-cells by endogenous mouse mammary tumor virus 3 prevents type 1 diabetes induction by autoreactive CD8+ T-cells
Authors: Driver John P , Ye Cheng , Clements Sadie , Gu Weihong , Geurts Aron , Mathews Clayton , Serreze David V , Chen Yi-Guang 
Published: August 2023

Abstract
In both humans and NOD mice, type 1 diabetes (T1D) develops from the autoimmune destruction of pancreatic beta cells by T-cells. Interactions between both helper CD4+ and cytotoxic CD8+ T-cells are essential for T1D development in NOD mice. Previous work has indicated that pathogenic T-cells arise from deleterious interactions between relatively common genes which regulate aspects of T-cell activation/effector function (Ctla-4, Tnfrsf9, Il2/Il21), peptide presentation (H2-Ag7, B2m), and T-cell receptor signaling (Ptpn22). In this study, we used a combination of subcongenic mapping and a CRISPR/Cas9 screen to identify the NOD-encoded Mtv3 provirus as a genetic element affecting CD4+/CD8+ T-cell interactions through an additional mechanism, altering the T-cell receptor (TCR) repertoire. Mtv3 encodes a superantigen that deletes the majority of Vβ3+ thymocytes in NOD mice. Ablating Mtv3 and restoring Vβ3+ T-cells has no effect on spontaneous T1D development in NOD mice. However, transferring Mtv3 to C57BL/6 (B6) mice congenic for the NOD H2g7 MHC haplotype (B6.H2g7) completely blocks their normal susceptibility to T1D mediated by transferred CD8+ T-cells transgenically expressing AI4 or NY8.3 TCRs. The entire genetic effect is manifest by Vβ3+ CD4+ T-cells, which unless deleted by Mtv3, accumulate in insulitic lesions and trigger in B6 background mice the pathogenic activation of diabetogenic CD8+ T-cells. Our findings provide evidence that endogenous Mtv superantigens can influence autoimmune responses. Furthermore, since most common mouse strains have gaps in their TCR Vβ repertoire due to Mtvs, it raises questions about the role of Mtvs in other mouse models designed to reflect human disorders.
Supported by grants from the American Diabetes Association (1-14-BS-051) and NIH (AI130656)
Generation of autoreactive CD8 T cell in a mouse model of alopecia areata
Authors: Lee You Jeong , Lee Hee Eun 
Published: August 2023

Abstract
Alopecia areata (AA) is the most frequent form of an organ-specific autoimmune disease in humans mediated by NKG2D+ CD8 T cells destroying hair follicles. C3H/HeJ mice naturally develop AA similar to humans, and we found it accompanies the expansion of exhausted phenotype (CD101+ Tim3+) memory CD8 T cells not only in the skin and skin-draining lymph nodes (SDLNs), but also in the liver and lungs. Before the disease induction, we found these CD8 T cells are activated in the lungs and liver, suggesting they are pre-activated in these organs. In the skin, expanded CD8 T cells additionally upregulated CD39, PD-1, and CTLA-4, which produced less IFNg than those in the SDLN, liver and lungs when stimulated. These features suggest that autoreactive CD8 T cells in the skin are exhausted by chronic antigenic stimulation. However, they did not upregulate TOX, a hallmark of T cell exhaustion. Intriguingly, CD44high NKG2D-expressing CD8 T cells appeared in the thymi of aged C3H/HeJ mice before disease induction. These cells express TBET and EOMES without overt expansion of PLZF-expressing T cells. Collectively these features suggest AA is mediated by autoreactive activation CD8 T cells that are not explained by previously known mechanisms.
This research was supported by the National Research Foundation of Korea (NRF-2022R1A2C1007692)
The role of ST8Sia6 in braking inflammation in Inflammatory Bowel Disease
Authors: Crotts Sydney B , Antonetti Regina , Shapiro Michael , Rajcula Matthew , Hammer Brady , Mai Elissa , Shapiro Virginia 
Published: August 2023

Abstract
Inflammatory bowel disease (IBD) including ulcerative colitis (UC) and Crohn’s disease (CD) affects between 1–3 million people in the United States and costs an estimated $11–28 billion in national healthcare costs. Patients present with symptoms including diarrhea, rectal bleeding, and fatigue. Characterization of molecules that regulate IBD is critical in understanding pathogenesis. ST8Sia6 is a sialic acid transferase that adds terminal a2,8 disialic acids to cell surface glycoproteins, and we have found that ST8Sia6 knockout (KO) mice exhibit increased spontaneous small bowel (SB) lamina propria CD4 TH1 polarization, increased inflammatory cytokine production, decreased IgA class switching of SB plasma cells, and alterations of the SB innate lymphoid cells, indicative of gut inflammation. Moreover, adult ST8Sia6 KO mice have shortened small and large bowels, likely due to baseline inflammation. These mice do not exhibit overt baseline symptoms, but ST8Sia6 KO mice exhibit increased weight loss and decreased survival when challenged with dextran sodium sulfate (DSS)-induced colitis, which was also enhanced when co-housed with wildtype (WT) mice. DSS-challenged ST8Sia6 KO mice exhibit increased histological damage and immune infiltration of the gut as compared to WT controls. Accelerated DSS-induced colitis is also observed when ST8Sia6-deficient bone marrow is transplanted into WT recipients, indicating that the effect is mediated specifically by immune cells. This data demonstrates a role for ST8Sia6 in regulating the host immune response to IBD, and further elucidating this role will aid in developing a mechanistic understanding of IBD pathogenesis and in understanding potential immunotherapies for the disease.
R01CA243545, 5T32DK124190-03