bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nitric oxide dictates the reprogramming of carbon flux during M1 macrophage polarization
Authors: Palmieri Erika M. , Baseler Walter A. , Davies Luke C. , Gonzalez-Cotto Marieli , Ghesquiere Bart , Fan Teresa W. M. , Lane Andrew N. , Wink David A. , McVicar Daniel W. 
Published: January 2023

Abstract
Classical pro-inflammatory activation of macrophages is characterized by profound intracellular metabolic changes, with increased glycolytic usage of carbon, away from Oxidative Phosphorylation (OXPHOS). We previously demonstrated that Nitric Oxide (NO) levels induced in Bone Marrow Derived Macrophages (BMDMs) from Wild Type (WT) mice are necessary and sufficient for the repression of OXPHOS. Here we demonstrate that NO is also responsible for the “break” in the mitochondrial TCA cycle and citrate accumulation during LPS/IFNγ stimulation; macrophages that lack NO maintain indeed substantial levels of Oxygen Consumption Rates (OCR) and TCA cycle intermediates. Carbon tracing experiments in the presence of U13C-glucose show almost undetected labelled α-ketoglutarate from citrate in WT but a conserved pattern of heavy carbon fate in Nos2−/− macrophages. Moreover we found that mitochondrial respiration elicited through citrate was decreased in WT M1 macrophages, but isocitrate was a full substrate for complex I-dependent OCR suggesting suppression of metabolism at mitochondrial Aconitase (ACO2). Consistent with this data, we found ACO2 enzymatic activity blunted in WT vs Nos2−/−. In addition we observed that M1 macrophages reroute pyruvate away from Pyruvate Dehydrogenase (PDH) in an NO dependent manner since only WT show halted flux through PDH. Surprisingly, we demonstrate this mechanism to be independent on the activation of Hif1α and its suggested effect on limiting acetyl-coA for the TCA. With these data together we hypothesize that NO orchestrates macrophage metabolism during inflammation inhibiting OXPHOS by blocking Krebs Cycle, therefore depriving of substrates the mitochondrial electron transport chain.
mTOR induces lysosome expansion by selective translation of lysosomal transcripts during phagocyte activation
Authors: Hipolito Victoria E. B. , Diaz Jacqueline A. , Tandoc Kristofferson V. , Saric Amra , Toposiviric Ivan , Botelho Roberto J. 
Published: January 2023

Abstract
The molecular mechanisms that govern and adapt organelle number, size, morphology and activities to suit the needs of many cell types and the conditions that a cell may encounter remain poorly defined. Lysosomes are organelles that degrade cargo from a variety of routes including endocytosis, phagocytosis and autophagy. Lysosomes have emerged as a signalling platform that senses and couples stress signals such as nutrient deprivation to regulatory kinase hubs like mTOR and AMPK to modulate metabolic activity. For phagocytes and antigen-presenting cells like macrophages and dendritic cells, lysosomes are a kingpin organelle since they are essential to kill and process pathogens, and present antigens. During phagocyte activation, lysosomes undergo a striking reorganization, changing from dozens of globular structures to a tubular network, in a process that requires the phosphatidylinositol-3-kinase-Akt-mTOR signalling pathway. Ultimately, lysosome tubulation is thought to promote pinocytic retention and antigen presentation. We show that lysosome tubulation is accompanied by a rapid boost in lysosome volume and holding capacity during phagocyte activation with lipopolysaccharides. Unexpectedly, lysosome expansion was paralleled with the induction of lysosomal proteins, which was independent of TFEB and TFE3, transcription factors known to scale up lysosome biogenesis. Instead, we demonstrate a hitherto unappreciated mechanism of lysosome expansion via mTOR-dependent increase in translation of mRNAs encoding key lysosomal proteins including LAMP1 and V-ATPase subunits. Collectively, we identified a mechanism of rapid organelle expansion and remodelling driven by selective enhancement of protein synthesis.
Sympathetic neural control of inflammation by ADRB2-mediated IL-10 secretion
Authors: Agac Didem , Estrada Leonardo , Farrar John David 
Published: January 2023

Abstract
The nervous and immune systems reciprocally regulate their functions through the release of chemical messengers. Norepinephrine (NE), a neurotransmitter released by catecholaminergic nerve endings, allows the sympathetic nervous system to communicate with immune cells through adrenergic receptors (ADR). We identified a novel role for the β2-ADR (ADRB2) in controlling inflammation. NE suppresses pro-inflammatory cytokine secretion from primary macrophages in response to multiple TLR agonists, and ADRB2 signaling enhances early induction of IL-10. We assessed the in vivo role of this pathway using a LPS endotoxemia model. ADRB2 −/− animals rapidly succumbed to a sub-lethal LPS challenge, which correlated with elevated serum levels of TNFα and reduced IL-10. The lethality in the ADRB2 −/− animals was rescued by administering exogenous IL-10. Additionally, the ADRB2-specific agonist salmeterol rescued wild-type animals from a lethal LPS challenge, which was reversed by neutralizing anti-IL-10 antibody. Additionally, ADRB2 −/− mice are unable to control inflammation models of infection and colitis. These observations suggest that ADRB2 signaling is critical for controlling inflammation through the rapid induction of IL-10. Through transcriptome analysis, the NR4A nuclear orphan family members were induced by NE, and the presence of several putative NR4A binding sites within the IL-10 promoter suggests that these factors may directly regulate IL-10 expression in response to ADRB2 signaling. Further studies will test the control of IL-10 expression by the NR4A family of transcription factors. Understanding this pathway will provide new insights into how the nervous and immune systems communicate through ADRB2 signaling.
Isoflavones increase radiation-induced apoptosis and modulate pro-inflammatory signaling in EA.hy926 endothelial cells.
Authors: Fountain Matthew Donovan , McLellan Laura Ann , Cannon Alexa C. , Hillman Gilda G. 
Published: January 2023

Abstract

Background
The therapeutic use of x-ray ionizing radiation (XIR) for cancer treatment is limited by off target injury to normal tissues resulting in organ-specific toxicities with perpetual inflammation and fibrosis. XIR causes different forms of cell death including apoptosis, necrosis, and mitotic catastrophe that can dictate the resulting inflammatory response. Our lab has shown that soy isoflavones (SIF) reduce XIR-induced inflammation and fibrosis of thoracic organs. We now investigated the effect of SIF and XIR on endothelial cell death in vitro, which could play a role in SIF mitigation of inflammatory response.

Methods
EA.hy926 endothelial cells were treated with or without 30μM Novasoy 400 SIF for 24 hours prior to 3 Gy XIR and assessed at various time points. Cells were utilized for cell viability assays, stained for g-H2AX, Annexin-V, and NF-κB-p65, and lysed for western blot analysis of proteins involved in apoptosis and inflammatory signaling.

Results
Endothelial cell viability was significantly decreased for cells treated with SIF+XIR at 72 hours compared to XIR alone (p &lt;0.05). Endothelial cells treated with SIF+XIR had increased staining of Annexin-V and g-H2AX staining at 48 hours compared to XIR alone (p &lt;0.05). Western blot analysis showed increases in pro-apoptotic caspases 9 and 3 as well as Bax, IL-1b, ICAM-1 and VCAM-1 after cell treatment with SIF+XIR.

Conclusions
SIF enhances endothelial cell susceptibility to XIR-induced pro-apoptotic pathways resulting in greater apoptotic cell death. These findings suggest a mechanism of SIF-mediated radioprotection caused by increased apoptosis of normal cells and could limit the uncontrolled XIR-induced inflammatory signaling leading to organ fibrosis.
Docosahexaenoic acid as a prophylactic treatment on particle-exposed Balb/c mice
Authors: Fletcher Paige , Hamilton Raymond F. , Pestka James J. , Holian Andrij 
Published: January 2023

Abstract

Rationale
There is a rising trend in the use of nanomaterials in various consumer products; increasing the risk of human exposures. Inflammation is a vital contributing factor to pulmonary diseases which can be triggered by exposure to airborne toxicants such as nanomaterials. However, there are no effective treatments for acute inflammation. One potential treatment shown to have anti-inflammatory effects is the dietary supplement docosahexaenoic acid (DHA); an omega-3 polyunsaturated fatty acid found in fish oil. This study investigated DHA as a prophylactic treatment for particle-induced inflammation.

Methods
Balb/c mice were fed a control diet or a 1% DHA-containing diet for five weeks. At week four the mice were anesthetized and oropharyngeal instilled with dispersion media (50 μl) or a highly inflammatory multi-walled carbon nanotube (FA21; 50 μg). One week after instillation lung histology, macrophage NLRP3 inflammasome activation (IL-1β release) and lung lavage fluid (LLF) levels of high-mobility group box 1 (HMGB1), protein levels and inflammatory cytokines were evaluated. DHA’s impact on macrophage phenotype was also examined.

Results
Macrophage production of IL-1β and LLF levels of HMGB1 and protein decreased in mice fed the DHA diet. Mice exposed to FA21 on the DHA diet had less pro-inflammatory M1 macrophage phenotype gene expression compared to the control diet-fed mice in lung tissue and decreased pro-inflammatory cytokines in the LLF. The lung tissue airway thickness was also decreased in the DHA-fed mice.

Conclusion
DHA could potentially be used as a prophylactic treatment since it was demonstrated to have a number of anti-inflammatory effects on Balb/c mice exposed to a common nanomaterial.
TCR affinity influences helper T cell differentiation by biasing dendritic cell interactions
Authors: Kotov Dmitri Ivanovich , Mitchell Jason S. , Pengo Thomas , Kotov Jessica Amy , Ruedl Christiane , Way Sing Sing Y. , Langlois Ryan A. , Fife Brian T. , Jenkins Marc K. 
Published: January 2023

Abstract
Naïve CD4+ T lymphocytes differentiate into various subsets when their TCRs detect microbial peptide-MHCII complexes presented by dendritic cells. However, the mechanism by which TCR signaling influences differentiation is unknown. We found that low affinity TCRs biased naïve T cells to become B cell helpers while high affinity TCRs promoted the phagocyte helper fate. We created software for high-throughput, multicolor, quantitative imaging called Chrysalis to determine if differentiation was influenced by the type of dendritic cell that the naïve T cell interacted with. Our results show that T cells with high affinity TCRs had prolonged interactions with XCR1+ dendritic cells and tended to become phagocyte helpers, while T cells with lower affinity TCRs preferentially interacted with SIRPα+ dendritic cells and tended to become B cell helpers. Thus, TCR affinity influences T cell differentiation by dictating interactions with distinct dendritic cell types.
Salt-Inducible kinases are critical regulators of terminal B-cell differentiation
Authors: Robinson Emily , Care Matthew , Tooze Reuben , Doody Gina 
Published: January 2023

Abstract
The generation of antibody-secreting plasma cells is a step-wise process involving initial B-cell activation and expansion, followed by the persistence of end-stage effector cells that reside in supportive niches. The transition through these stages requires reprogramming for high levels of secretion and adaptation to the bioenergetic demands of producing immunoglobulin. To improve our understanding of the pathways involved, we applied gene co-expression network analysis to temporal data derived from in vitro differentiating human plasma cells. We identified salt-inducible kinase 1 (SIK1) as a highly connected hub gene involved in the transition from a cycling plasmablast to a quiescent plasma cell. SIK1 is part of a kinase family previously linked to the control of metabolism and the negative regulation of TLR signaling, in part through the inactivation of cAMP-regulated transcriptional co-activators (CRTCs). To assess the contribution of SIK family kinases during plasma cell differentiation, we utilized the SIK inhibitors HG-9-91-01 and YKL-05-099. In contrast to macrophages where SIK inhibition enhances anti-inflammatory polarization, exposure of primary human or murine B-cells that have activated plasma cell differentiation to the SIK inhibitors lead to a profound reduction in cell viability. These effects were also replicated in a subset of multiple myeloma cell lines. The results are consistent with the described requirement for CRTC2 inactivation to exit the germinal center reaction and initiate plasma cell differentiation. Our findings extend these earlier observations and provide scope for identification of additional mechanisms that control plasma cell quiescence and secretory capacity.
Ocular exocrine glands are infected, harbor T cells, and express select cytokines and chemokines following corneal HSV-1 infection.
Authors: Montgomery Micaela Lee , Carr Daniel J. J. 
Published: January 2023

Abstract
Herpes simplex virus (HSV)-1 is a pathogen that infects a large portion of the human population and establishes a life-long, latent infection. Reactivation of HSV-1 can manifest as “cold sores” or more severe pathologies, including corneal herpes stromal keratitis. The potential involvement of ocular glands following corneal HSV-1 infection is not well described, despite their significant contribution to eye health. CD-1 outbred mice were ocularly infected with HSV-1, and ocular exocrine glands, including harderian glands (HGs) and extraorbital lacrimal glands (ELGs), were evaluated for susceptibility to infection and immunological activity. HSV-1 was found to infect ocular glands in a time-dependent fashion with HGs infected earlier than ELGs. Surgical removal of HGs prior to infection resulted in a significant increase in infectious virus recovered from corneas, but not ELGs or brainstems, when compared to mice with intact HGs (p≤0.01). Bidirectional flow was shown to occur between ocular surface contents and HGs in less than 4 hours, which might contribute to HSV-1 trafficking to HGs. CD4+ and CD8+ T cell infiltration/expansion occurs within HGs and ELGs in a time-dependent fashion post-infection (pi) compared to B cells that began to infiltrate HGs, but not ELGs, by day 5 pi. Cytokines, including IL-4, IL-6, IL-22, and IFN-γ, were found to be significantly (p≤0.05) elevated in ELGs, but not HGs, at day 7 pi. Chemokines, including CCL2, CCL5, and CXCL10, were significantly (p≤0.05) elevated in both ELGs and HGs at day 7 pi. These results underscore a dynamic response of ocular exocrine glands to corneal HSV-1 infection and the potential contribution these glands make to viral resistance.
Deficiency of neutrophil extracellular traps increases susceptibility to colonic inflammation
Authors: Singh Vishal , Yeoh Beng San , Xiao Xia , Wang Yanming , Vijay-Kumar Matam 
Published: January 2023

Abstract
Neutrophil extracellular traps (NETs), DNA structures released from neutrophils, are primarily enriched with bactericidal proteins. We envision that the antimicrobial nature of NETs would be an appropriate immune response in the gut, which is continuously exposed to trillions of bacteria. NETs formation is peptidyl arginine deiminase-4 (PAD4)-dependent, therefore, we employed Pad4KO mice and their WT littermates in two well-established models of murine colitis, i.e., loss of IL-10 signaling (immune hyperactivation) and chemical-induced injury (DSS). Upon treatment with αIL-10R mAb, Pad4KO mice developed robust chronic colitis as exemplified by colomegaly, colonic crypt elongation, and increased immune cell infiltration. In comparison, αIL-10R-treated WT mice exhibited modest colitis. We theorized that NETs may function as a ‘safety net’ which confine the pro-inflammatory neutrophil granule proteins (NGP) and prevent them to cause further damage. Indeed, the concentration and activity of NGP [i.e. lipocalin 2 (Lcn2), myeloperoxidase (MPO) and neutrophil elastase (NE)], in serum and in colons, were strikingly elevated in colitic Pad4KO mice, when compared to WT mice. The level of colonic secretory leukocyte protease inhibitor (SLPI) was reduced in the colitic Pad4KO mice. Such impaired SLPI response coupled with the ‘spillover’ of NGPs may, in part, explain the aggravated colitis in mice lacking NETs. Similarly, upon DSS treatment Pad4KO mice developed more severe chronic colitis than WT mice. Taken together, our findings advance the notion that gastrointestinal NETs are beneficial, and elucidation of NETs-mediated signaling, during colitis, may yield a novel therapeutic approach to mitigate intestinal inflammation.
Resveratrol increases efferocytosis via both Mertk-GAS6/PROS and Integrin-TG2-Mfge8 pathway
Authors: Tsay Gregory J. 
Published: January 2023

Abstract
The impairment of clearance of apoptotic cells (efferocytosis) leads to local inflammatory response and tissue damage. Recent studies have found the defective mechanism in apoptotic clearance is related to the pathogenesis of autoimmune disease such as systemic lupus erythematosus (SLE).
There have been many studies about the positive effects of resveratrol, including anti-diabetic effect, anti-inflammation, anti-oxidation, and anti-aging. However, the effect of resveratrol on clearance of apoptotic cells by macrophage is still remaining unknown. We investigated the effects of resveratrol on efferocytosis.

Methods
HaCaT and the thymocytes from C57BL/6 were induced apoptosis by UVB and stained with Deep red mitotracker. CFSE stained RAW 264.7 was treated with 1, 5, 10 mM resveratrol for 24 hrs then co-cultured with apoptotic HaCaT to investigate the effect of resveratrol on efferocytosis. To measure the mRNA expression, RAW 264.7 RNA was extracted by RNAzol and then progress reverse transcription and real time PCR.

Results
The results showed that 10 mM resveratrol increased efferocytosis. We further investigated the molecular mechanism of efferocytosis. We found that ITGa5, ITGb3, TG2, Mfge8, and Mertk mRNA were up-regulated by resveratrol after co-cultured with or without apoptotic HaCaT; GAS6 and PROS mRNA expression were up-regulated by resveratrol only after co-cultured with apoptotic cells. The results revealed that resveratrol might enhance efferocytosis by regulating Mfge8-Integrin and GAS6/PROS-Mertk signaling pathway.

Conclusion
Resveratrol enhances efferocytosis via both Mertk-GAS6/PROS and Integrin-TG2-Mfge8 pathway. Resveratrol may be used for the therapy of autoimmune diseases.
Modulating T cell function through elongation of the mitochondrial network to improve cytotoxic CD8+ tumor infiltrating lymphocyte function
Authors: Gonzales Marissa A. , Bullock Timothy N. J. 
Published: January 2023

Abstract
For a number of tumor types, including melanoma, positive clinical outcomes correlate with infiltration of T-cells into the tumor. However, failure to completely control disease progression often occurs, even with immune responses enhanced by checkpoint blockade or other immunotherapy. When T-cells fail to control tumor growth, the tumor infiltrating T lymphocytes (TIL) exhibit characteristics of exhaustion such as poor proliferation, poor effector function, and an increase in inhibitory receptor expression. Metabolic activity of T-cells contributes to differentiation and effector function, leading us to investigate the metabolic state of TIL. By flow cytometry, we find that CD8+ cytotoxic TIL have lower mitochondrial mass, as well as lower mitochondrial membrane potential, compared to effector CD8+ T-cells that respond to vaccination. We hypothesize that TIL are unable to effectively regulate their mitochondrial network, and it is likely that one or more fusion or fission machinery proteins has altered expression or can be manipulated to improve TIL function. We find that TIL have punctate mitochondria as compared to functional effector cells and hypothesize that elongation of the mitochondrial network will improve TIL function such as increasing the capacity for OXPHOS, which is seen in memory T cells with elongated mitochondrial networks. Using mice lacking the fission protein, Drp1, in T cells as a model of elongated mitochondria, we see increased mitochondrial membrane potential, spare respiratory capacity and OXPHOS as compared to WT cells with fission capability. We continue to investigate the effects of mitochondrial elongation of CD8+ cytotoxic TIL.
T cell co-stimulation reprograms the metabolic fitness of highly differentiated CD8+ T cells for improved adoptive T–cell therapy
Authors: Srivastava Abhishek K. , Paria Biman C. , Chandran Smita , Kammula Udai S. 
Published: January 2023

Abstract
Adoptive T cell therapy (ACT) using tumor infiltrating lymphocytes (TIL) can result in durable complete cancer remission in select patients with metastatic melanoma. These clinical responses have strongly correlated with the ability of the transferred cells to engraft and persist in these patients. Preclinical murine studies have further supported that T cells with superior anti-tumor efficacy have higher proliferative capacity, reduced AICD and enhanced in vivo persistence. Thus, in an effort to further improve ACT, we sought to exploit ex vivo 4-1BB co-stimulation to generate T cells with enhanced in vivo anti-tumor activity. We used an artificial APCs genetically engineered to express 4-1BBL to provide ex-vivo 4-1BB co-stimulation to human tumor specific CD8+ T cells. Our results demonstrate that ex-vivo 4-1BBL co-stimulation enhanced proliferative capacity and reduced AICD of the CD8+ T cells without loss of functional tumor avidity. Mechanistically, 4-1BB co-stimulation demonstrated enhanced glycolysis (ECAR) and mitochondrial respiration (OXPHOS) further suggested that 4-1BB co-stimulation could induce significant reprogramming of the bioenergetic pathways used by highly differentiated T cells. The 4-1BB co-stimulation further restored the ability of the highly differentiated T cells to persist and survive after adoptive transfer in NSG mice and resulted in improved survival of established tumor bearing NSG mice. These findings shed light on a fundamental role of 4-1BB co-stimulation and further suggest a practical approach to reprogram the metabolic fitness of highly differentiated T cells for adoptive immunotherapy