bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hippuric acid inhibits survival of germinal center B cells via GPR109A in kidney disease
Authors: Peroumal Doureradjou , Jawale Chetan , Antos Dani , Li De-Dong , Choi Wonseok , Wang Shuxia , Alcorn John , Biswas Partha 
Published: November 2024

Abstract
The formation of germinal center (GC) response is crucial for antibody mediated immunity against infection. However, compromised humoral immunity to infection was seen in patients having pre-existing comorbid conditions including kidney disease. The mechanisms by which kidney disease account for poor antibody response is unknown. Here we show that kidney disease negatively impacted GC formation, antibody affinity maturation, isotype switching and titer. GC B cells exhibit increased apoptosis in kidney disease. Uremic toxin hippuric acid drives loss of mitochondrial membrane potential leading to increased apoptosis of GC B cells in G-protein–coupled receptor109A (Gpr109A) dependent manner. GC B cells express Gpr109A and mice without kidney disease treated with Gpr109A agonist exhibit similar defect in GC formation, affinity maturation and antibody titer. Conversely, immunized Gpr109A global knock-out or mice in which B cells are lacking Gpr109a show no defect in GC formation, affinity maturation and antibody titer following kidney disease. These results shed lights on hippuric acid/Gpr109A axis in causing GC B cells apoptosis and provide a mechanistic understanding of how renal dysfunction suppresses humoral immunity in patients with kidney disease.
Investigating Enzymatic Control of Inflammation Oxo-eicosanoid Synthesis in Live Zebrafish
Authors: Ma Yanan , Hui King Lam , Niethammer Philipp , Gelashvili Zaza , Lengyel Miklos 
Published: November 2024

Abstract
Eicosanoids, derived from arachidonic acid (AA), are bioactive lipids with chemokine-like function. While prostaglandin and leukotriene have been extensively studied, the genetics of oxoeicosanoid metabolism remain relatively unexplored. The proto-type oxoeicosanoid 5-KETE is generated from AA by 5-lipoxygenase and the long-elusive redox-sensitive enzyme 5-HEDH. Pure 5-KETE attracts leukocytes in vitro and in vivo via the G-protein coupled receptor OXER1. 5-KETE is induced in response to Ischemic, allergic challenges, and colitis. OXER1 inhibition suppresses neutrophilic inflammation in zebrafish and primate asthma models and frameshift mutations of OXER1 in mucosal tumors suggest a yet undescribed cancer role. However, the lack of OXER1 orthologs in rodents and uncharacterized 5-HEDH enzyme have severely stalled oxoeicosanoid research for the past 30 years. Through a combination of lipidomics, enzymatic activity screening and genetic perturbation in human cell culture, coupled with intravital imaging of acute inflammation in zebrafish, we here identify the first-in-class 5-HEDH member. Its genetic deletion severely compromises 5-KETE synthesis in human cells and abrogates rapid neutrophil and macrophage recruitment to zebrafish tail fin wounds in vivo, indicating its role in neutrophil and macrophage interplay. Our work will open new therapeutic avenues for treating acute and chronic mucosal inflammation.
Aryl hydrocarbon receptor control of α4β7 and chemokine receptor 9 expression on B cells is partially determined by sex
Authors: Bauerle Logan , DeKrey Gregory 
Published: November 2024

Abstract
The aryl hydrocarbon receptor (AhR) is a ligand activated transcription factor that regulates immune function differentially by sex. In this study, we compared the impact of AhR activation on the expression of chemokine receptor (CCR) 9 and α4β7 integrin on male and female B cells in vitro. Splenic B cells were isolated from C57Bl/6J mice and then stimulated in culture with 300 ng/mL anti IgD-dextran, 2 ng/mL human TGF-β, and 25 μg/mL LPS. Cells were also exposed to either 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, agonist, 50 nM) or CH223191 (100 μM, antagonist). After 96 hours, cells were analyzed by flow cytometry. Under antagonist conditions, the percentage of viable cells expressing CCR9 was 3.8% (male) and 5.2% (female), and α4β7 expression was 1.5% (male) and 1.6% (female) (N=6). AhR activation significantly increased (P &lt; 0.05) the percentage of viable cells expressing either CCR9 or α4β7 by approximately 3-fold for male cells and &gt;4-fold for female cells. Relative to males, a significant increase in the proportion of female cells expressing CCR9, but not α4β7, occurred following AhR agonism. The level of CCR9 expressed on individual cells (mean fluorescence intensity) was also significantly increased by AhR activation (approximately 50%, log scale) in both males and females, but the level of α4β7 expression was significantly increased only in female mice. These results suggest that AhR activation may enhance B cell migration into the gut preferentially in females.
Aryl hydrocarbon receptor activation ameliorates colitis via transcriptional regulation of -defensin1
Authors: Palrasu Manikandan , Kakar Khadija , FNU Hamida , Marudamuthu Amarnath Satheesh , Carter Tayler , Wilson Kiesha , Saxena Archana , Yang Xiaoming , Singh Narendra , Busbee Brandon , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: November 2024

Abstract
Ulcerative colitis (UC), a chronic inflammatory disease of the colon and rectum accounts for 1.5 million individuals, with an annual incidence of 15 per 100,000 individuals. The etiology of colitis includes host, genetic, environmental, and microbiological factors. The intestinal epithelium can produce antimicrobial molecules for epithelial protection from commensal microorganisms. Aryl hydrocarbon receptor (AhR) plays an important role as an environmental sensor and a regulator of the immune response. Indeed, studies have shown that the expression of AhR was correlated with the expression of a-defensins in inflammatory bowel disease patients. Therefore, the present study was designed to investigate how AhR-ligand activation regulates a-defensin1 in a colitis model. Here, we discovered that in intestinal epithelial cells from colitis mice, dietary Indole-3-carbinol (I3C), a ligand for the AhR, induced AhR and α-defensin1. We also found that AhR-null animals, which are more likely to develop colitis, did not exhibit an increase in α-defensin1 following I3C treatment, indicating that AhR regulates α-defensin1. Similar findings were observed in MC38 cells. Furthermore, the DRE region in the a-defensin1 promoter was involved in the upregulation of a-defensin1 by AhR upon I3C treatment. Collectively, these results provide evidence that AhR activation followed by a-defensin1 production may be a vital prophylactic and therapeutic measure for colitis.
Perturbation of azurophilic granule integrity drives NLRP3-independent IL-1b processing and release in neutrophils
Authors: Miller Brandon , Dubyak George 
Published: November 2024

Abstract
IL-1β is an inflammatory cytokine secreted by myeloid cells in response to infection or sterile tissue damage. Non-canonical secretion of IL-1β from monocytes downstream of activated NLRP3 inflammasomes is the best-characterized model; this is mediated by caspase-1 cleavage of GSDMD and N-GSDMD plasma membrane (PM) pore formation to permit release of IL-1β and induction of pyroptosis. NLRP3 is a cytosolic sensor of cellular homeostasis including [K+]. In monocytes, K+ efflux mediated NLRP3 activation is triggered by agents that disrupt lysosomal integrity. Neutrophils assemble competent NLRP3 inflammasomes and release IL-1β via GSDMD-dependent mechanisms, but they resist N-GSDMD PM pore formation and pyroptosis. We tested whether lysosome-disrupting stimuli in neutrophils would phenocopy the monocyte mechanism of NLRP3-dependent IL-1β release. Surprisingly, our data indicates that lysosome disruption induces release of mature IL-1β and lytic cell death in an NLRP3-independent manner. Kinetic analysis demonstrates an alternate mechanism of release such that early phase IL-1β release is NLRP3-dependent, but prolonged stimulation leads to NLRP3-independent release. Collectively, our data supports a signaling mechanism by which azurophilic granule disruption and cytosolic accumulation of neutrophil serine proteases disrupt canonical NLRP3 inflammasome assembly and directly cleave proIL-1β as part of a novel neutrophil-specific signaling mechanism of IL-1β processing and export.
Single-cell proteomic analysis of peripheral blood mononuclear cells in patients with pulmonary long COVID
Authors: Guinto Elizabeth , Kuo Grace , Messing Melina , Cheung Chung , Yip Will , Gerayeli Firoozeh , Eddy Rachel , McNagny Kelly , Sin Don 
Published: November 2024

Abstract
An estimated 15% of patients are expected to continue to exhibit respiratory symptoms after their SARS-CoV-2 infection. The mechanisms behind these symptoms remain unknown. We recruited twenty-three patients (65% female) who were on average 406 ± 251 days post-COVID-19. The cohort consists of primarily mild (non-hospitalized) patients (87%), with an average age of 43 ± 15 years. Nine of these patients had self-reported dyspnea, which we defined as pulmonary long COVID (PLC). The remaining 14 patients recovered without persistent dyspnea. We performed a 38-panel cytometry by time-of-flight on the peripheral blood mononuclear cells of this cohort. Quality control was performed using FloJo. Further analysis, was done using the CyCombine, CATALYST, and FlowSom packages in R. Our final annotation uncovered 17 different cell clusters. In PLC patients, double-positive T-cells (DPTs) were significantly increased (Wilcoxon p&lt;0.05), indicating that T-cell development or differentiation may be altered in these patients. Though DPTs are known to be increased in females, there was no significant difference in the proportion of DPTs between males and females in our cohort. PLC stem cells had increased expression of CD304 and CD200R (Wilcoxon p&lt;0.05). CD304 is as a co-receptor for SARS-CoV-2, while CD200R is a limiter of pro-inflammatory activity. These findings indicate that the immune landscape in PLC patients is altered, compared to those who recover without residual pulmonary symptoms.
Gpx4 regulates iNKT cell homeostasis and function by preventing ferroptosis and lipid peroxidation
Authors: Sok Sophia P M , Pipkin Kaitlyn , Van Remmen Holly , Fan Zhichao , Zhao Meng 
Published: November 2024

Abstract
Invariant natural killer T (iNKT) cells are a group of innate-like T cells recognizing lipid Ags and plays important roles in immune responses. We found iNKT cells, compared to CD4T cells, have significantly higher level of lipid peroxidation, lower labile iron pool, and higher level of Gluthathione peroxidase 4 (GPX4), a major antioxidant enzyme protecting cells from ferroptosis. T cell-specific KO of Gpx4 reduced iNKT cells, especially NKT1 cells, the IFNg producing subset. RNAseq revealed Th1 pathway, cell cycle, and mitochondrial function are perturbed by Gpx4 deletion in iNKT cells. Indeed, we detected impaired cytokine production, elevated cell proliferation and cell death, accumulation of lipid peroxides and mitochondrial ROS in Gpx4 KO iNKT cells. Ferroptosis inhibitor, iron chelator, vitamins E, and mitochondrial antioxidant (MitoQ) prevented the ferroptotic cell death induced by Gpx4 inhibition in iNKT. Moreover, the function of the Gpx4 KO iNKT cells can be restored by antioxidants or iron chelator. Notably, vitamin E rescued iNKT cells at homeostasis in Gpx4KO mice. Furthermore, vitamin K2 prevented the cell death and restored the cytokine production in Gpx4-cKO iNKT cells in vitro, and it also recovered the percentage of iNKT1 in Gpx4 cKO mice when administrated in vivo, consistent with a protective role of vitamin K cycle during ferroptosis.
Complement C4 and Autoimmune Diseases
Authors: Zhou Danlei , Coss Samantha , Abdul Aziz Rabheh , Drew Joanne , Miller Katherine , Driest Kyla , Sivaraman Vidya , Spencer Charles , Barbar-Smiley Fatima , Ardoin Stacy P , Yu Chack-Yung 
Published: November 2024

Abstract
Complement activation on autologous cells causes tissue injuries but the process is not well studied. We seek to elucidate how C4 variants, their activation products and isotype deficiencies contribute to increased risks of children systemic lupus erythematous (cSLE) and juvenile dermatomyositis (JDM). We performed kinetic studies of C4A and C4B reactivities by measuring hemolysis of sensitized sheep erythrocytes (EA). EDTA-plasma diluted to 1% showed that C4B lysed EA immediately, as revealed by released hemoglobin at OD415 nm. Those of C4A exhibited hysteresis property. C4A probably mitigates damaging effects of activated C4B on host cells. We performed flow cytometry to evaluate cell-bound complement split products on blood cells using specific monoclonals for samples with cSLE (n=119), JDM (n=159), and controls (HC, n=109). Non-parametric analyses for median fluorescent intensities were compared with median and IQR presented. The three sets of data among cSLE, JDM and HC were 795 (523, 1161), 350 (181, 779) and 252 (163, 252), respectively, for E-C4d (cSLE/HC, p=3.8x10-22; JDM/HC, p=8.8x10-8); 201 (95, 328), 33 (4, 101), and 26 (6, 70), respectively, for E-C4Ad (p=5.3x10-10, cSLE/HC; p=0.02, JDM/HC); and 68 (30, 154), 11.9 (1.9, 23.8), and 13.5 (8, 19.2) for E-C3d (p=1.8x10-6, cSLE/HC; p=0.16, JDM/HC). In conclusion, complement activation on host cells is retarded after activation of C4. Moreover, activation of C4 is substantially higher in cSLE and JDM than HC.
Proton therapy inhibits the production of immune suppressive exosomes in head and neck squamous cell carcinoma
Authors: Chimote Ameet , Lehn Maria , Bhati Jay , Mascia Anthony , Sertorio Mathieu , Lamba Michael , Ionascu Dan , Tang Alice , Langevin Scott , Khodoun Marat , Wise-Draper Trisha , Conforti Laura 
Published: November 2024

Abstract
Proton therapy (PT) is a promising new alternative to conventional X-ray-based photon therapy (XRT) for patients with advanced head and neck squamous cell carcinomas (HNSCC) owing to its precise dosimetric characteristics and reduced toxicity. While tumor-derived exosomes contribute to immune suppression in HNSCC, the effects of PT on tumor-derived exosomes and anti-tumor immune responses are poorly understood. In this study, we generated primary HNSCC cell cultures from resected tumors and irradiated them with 5 Gy PT or XRT, and isolated exosomes from cell culture supernatants. We observed that compared to XRT, PT-exposed HNSCC cells produced 75% fewer exosomes. To assess the immunomodulatory effects of PT, we exposed peripheral blood mononuclear cells (PBMCs) to exosomes from PT- and XRT-irradiated HNSCC cells and conducted preliminary single-cell studies by transcriptome analysis (NanoString), secretome profiling (Isoplexis) and immune function (IFN-g release). Transcriptomic studies revealed that exosomes from PT- and XRT-irradiated cells comparably reduced cytotoxic and Th1 cell abundance, and inhibited immune responses associated with cytotoxicity, NK, and T cell functions. Exosomes from PT- and XRT-irradiated cells equally suppressed IFN-g release and reduced the polyfunctionality (the ability to secrete multiple cytokines from a single cell) of CD8+ T cells. Our results suggest that PT limits the immune suppressive effect of exosomes by reducing their production.
Postprandial changes to systemic metabolism imprint durable changes on T cell immune responses
Authors: Kumar Alok , Rivadienera Dayana , Delgoffe Greg , Poholek Amanda , Kohan Alison 
Published: November 2024

Abstract
Here we show short periods of fasting and refeeding can have long-lasting effects on T cell immunity. Mouse or human T cells from fed hosts have higher mitochondrial quality and respiratory capacity than those from fasted hosts ex vivo. These metabolic phenotypes persist after activation and expansion, both in vitro culture and in vivo in response to vaccination. Fed T cells outcompete fasted T cells in a co-transfer model, such that T cells from fasted hosts form far fewer memory T cells. Serum profiling revealed triglycerides as a driver of postprandial metabolic reprogramming: chylomicrons enriched from the lymphatics of fed mice were sufficient to enhance mitochondria of fasted T cells. LDLR-deficient T cells had the metabolic phenotype of fasted T cells and were insensitive to fed serum, confirming the role of triglyceride uptake in postprandial T cell metabolic programming. Transcriptomic analysis revealed higher expression of LDLR, OXPHOS proteins, and fatty acid metabolism enzymes in fed T cells over fasted T cells. Finally, human CAR T cells manufactured from the same donor after a meal show a therapeutic advantage over T cells collected in the fasted state. In summary, postprandial metabolism imparts durable metabolic and functional advantage on T cells. Our study highlights the need to consider diet content and timing as key factors in immune cell analysis, vaccination strategies, and the generation of cellular therapies for cancer.
Electrical stimulation of the arterial splenic nerve has durable therapeutic efficacy on murine collagen-induced arthritis.
Authors: BONDUE Alexandre 
Published: November 2024

Abstract
Vagus nerve (VN) stimulation has anti-arthritic effects in animal models and in patients with rheumatoid arthritis (RA). However, VN stimulation may affect several physiological functions. The splenic nerve (SpN) has been implicated in VN-mediated immunomodulation. We tested the therapeutic efficacy of arterial SpN stimulation (SNS) on collagen-induced arthritis (CIA) in mice as a single treatment or in combination with etanercept or Jak inhibitor. SNS was evaluated in mice with CIA before or after disease onset and applied one or six times daily. Alternatively, mice were treated with etanercept or propranolol three times a week. SNS treatment of CIA mice before the onset of clinical symptoms prevented disease in a β1/β2-adrenoceptor-dependent manner. After SNS treatment, splenic inflammatory monocytes, B cells and autoantibody-secreting cells were reduced, as were serum levels of TNF and anti-collagen antibodies. When SNS was tested as a therapeutic modality in already diseased mice, clinical scores were significantly reduced compared to sham-treated animals. Importantly, the therapeutic efficacy of SNS was enhanced when combined with etanercept treatment. The results of this study suggest that SNS may have potent therapeutic anti-arthritogenic effects.
Efficacy of a novel oral, potent, and highly selective PKCt inhibitor in the SCID T-cell transfer colitis model
Authors: Harden Jamie , Whitener Robert , Kim Andrea , Hofland Hans , Patel Jeegar 
Published: November 2024

Abstract
Protein kinase C family theta isoform (PKCt) is a key kinase downstream of T-cell receptor (TCR) and CD28 co-signaling in T-cells, leading to T-cell activation and production of effector cytokines. As T-cells are key pathogenic cells in a multitude of chronic inflammatory diseases, inhibition of PKCt with an oral small molecule may provide a novel mechanism to treat chronic inflammatory diseases, such as colitis. In this study, we evaluated the efficacy of two orally bioavailable, potent, and highly selective PKCt inhibitors in the severe-combined-immunodeficient (SCID) T-cell transfer colitis model; this model is considered the most relevant to human inflammatory bowel disease (IBD), and displays characteristics of both Crohn’s disease (CD) and ulcerative colitis (UC). PKCt inhibition resulted in a dose-dependent prevention of colitis symptoms, including maintenance of body weight and stool score, with efficacy similar to an anti-IL-12 monoclonal antibody. Histological assessment and tissue cytokine and chemokine protein quantification of the colon were also performed. These data suggest that an oral PKCt inhibitor may have utility in the treatment of IBD, and other chronic inflammatory diseases driven by pathogenic T-cell activation.
Efficacy of lymphatic pump treatment in conjunction with methotrexate on rats with adjuvant-induced arthritis
Authors: Volin Michael , Patel Saahil , Zanotti Brian , Monroy Del Toro Rosalinda , Incrocci Ryan , Swanson-Mungerson Michelle 
Published: November 2024

Abstract
Rheumatoid Arthritis (RA) is an autoimmune disease targeting synovial joints in which self-reactive T cells release cytokines and B cells produce self-reactive antibodies resulting in an inflammatory response. Lymphatic pump treatment (LPT) promotes lymphatic flow resulting in a flushing out of the immune-complexes and cytokines. A rat mode on RA (AIA) was used to determine whether LPT in conjunction with the anti-rheumatic drug, methotrexate (MTX), enhanced its therapeutic impact on RA. A preventative study was performed in which MTX was given along with LPT (3 times a day for 7 days) . Ankles were assessed daily for signs of inflammation. Flow cytometry was performed on spleen and popliteal lymph nodes (PLN) to determine changes in leukocyte populations. Cytokine levels were determined on ankle lysates. The study showed that MTX treatment delayed the onset of arthritis. MTX reduced the spleen weights (p&lt;0.05). LPT also reduced spleen weights but did not enhance the MTX reduction. LPT reduced PLN weights compared to MTX. MTX resulted in higher numbers of CD4+ T cells and lower numbers of CD8+ T cells and B cells in spleens. PLNs treated with MTX had reduced lymphocyte numbers. MTX treatment reduced CINC-1 and TNFa levels (p&lt;0.05). LPT also reduced CINC-1 levels. In conclusion, MTX treatments delayed arthritis formation in AIA rats while LPT affected both spleen and PLN weights. Thus, the addition of LPT to RA treatment showed potential benefits without negative effects.
Joint single-cell transcriptomic and epigenomic analysis reveals key regulators of CAR T cell stemness and antitumor immunity
Authors: Wu Tuoqi , Zhu Ziang , Wizzard Safuwra , Yao Chen 
Published: November 2024

Abstract
The differentiation of antitumor T cells is tightly controlled by transcriptional and epigenetic pathways. Understanding the programs governing the differentiation of therapeutic T cells such as chimeric antigen receptor (CAR) T cells is crucial for improving their efficacy. Using a single-cell multi-omics approach, we simultaneously profiled chromatin accessibility and transcription within the same single CAR T cell in mice with B-cell leukemia or melanoma. One of the CAR T-cell subsets displayed stem cell-like properties and greater proliferative potential upon tumor challenge. In addition, we uncovered primed chromatin states foreshadowing transcriptional changes and lineage-defining genes heavily regulated at the epigenetic level. By constructing gene regulatory networks in CAR T cells based on the single-cell multi-omics data, we identified hub transcription factors of stem-like CAR T cells including BACH2. By knocking out BACH2 in CAR T cells, we demonstrated an essential role of BACH2 in the development of the stem-like subset and antitumor immunity of CAR T cells. Thus, our study elucidated the transcriptional and epigenetic mechanisms underlying CAR T-cell differentiation and revealed key transcriptional regulators controlling the differentiation and antitumor immunity of CAR T cells.