Minimizing unintended granulocyte activation while measuring functional responsiveness is essential, as the use of external probes, antibodies, or fluorescent dyes can potentially alter cellular responsiveness. To address this, we employed an antibody-free flow cytometry approach that measures forward scatter (FSC) to detect variations in cell-size, morphology, and shape; some key indicators of neutrophil and eosinophil activation. Human peripheral blood neutrophils, containing contaminating eosinophils, were isolated using discontinuous Percoll gradients and pre-treated with receptor antagonists [e.g., cyclosporin-H (an FPR1 antagonist) and CP105696 (a BLT1 receptor antagonist)] prior to stimulation with agonists such as fMLF (an FPR1 agonist) and LTB (a BLT1 agonist). Furthermore, fMLF stimulation resulted in a loss of CD62L and an increase in CD11b expression along with an increase in intracellular ROS production compared to control, as analysed using flow cytometry. Imaging flow cytometry, together with FSC analysis, enabled assessment of cell polarization and associated morphological changes. Importantly, autofluorescence-based gating allowed for the identification of contaminating eosinophils within the mixed granulocyte population, allowing parallel assessment of shape-change in both neutrophils and eosinophils in response to the same ligands. Stimulation of neutrophils with fMLF resulted in distinct FSC shifts compared to unstimulated controls across all flow cytometers tested, which were inhibited by cyclosporin-H, but not CP105696. Morphological analysis confirmed these changes corresponded with increased cell area and perimeter and decreased circularity, hallmarks of cell polarisation. Additionally, selective activation of eosinophils (but not neutrophils) by eotaxin, and dual activation of both cell types by the arachidonic acid metabolite 5-oxo-ETE, were confirmed through specific gating strategies. Taken together, these findings support the use of FSC-based flow cytometry as a rapid, scalable and effective method for evaluating granulocyte polarisation and screening candidate therapeutics targeting immune cell activation in disease contexts.

Chronic obstructive pulmonary disease (COPD) is characterized by inflammation and an immune response. However, the relationship between ferroptosis and COPD remains unknown. We aim to identify pivotal ferroptosis-related biomarkers in COPD and explore their roles in immune infiltration landscapes.

Phosphorylation of eIF4E by MNK1/2 modulates protein synthesis by controlling the translation of specific mRNAs. Immune cells use the MNK1/2-eIF4E axis to adapt their gene expression in response to environmental cues, but its dysregulated activity promotes disease progression. While recent examples using cancer models have identified CD8 T-cells as a conduit for the tumor-supporting role of the MNK1/2-eIF4E axis, the impact of phospho-eIF4E on CD4 T-cell subsets, specifically regulatory T-cells, remains unclear. To fill this knowledge gap, we studied the impact of phospho-eIF4E-deficiency on Treg activity in mice in an inflammatory context using a model of murine colitis and in human PBMCs.

This study aimed to investigate the role of butyrate in regulating STING-induced endoplasmic reticulum stress (ERS) and CD4 tissue-resident memory (TRM) T cells responses during the progression of ulcerative colitis (UC). Our results demonstrated that butyrate significantly alleviated dextran sulfate sodium (DSS)-induced colitis, as evidenced by restored intestinal epithelial architecture, reduced inflammatory cytokine, and decreased CD4 TRM T cells. These protective effects were likely mediated through modulation of the STING-ERS pathway. Using a CT26 cell model, we further confirmed that STING activation promotes ERS, leading to enhanced secretion of inflammatory factors and subsequent induction of CD4 TRM T cells. Importantly, butyrate effectively suppressed this STING-initiated inflammatory cascade in intestinal epithelial cells (IECs). Our findings revealed a novel mechanism by which butyrate ameliorates UC through inhibition of the STING-ERS axis in IECs, highlighting its therapeutic potential for UC treatment.

Emerging evidence suggests that renal inflammation, characterized by the activation of renal macrophages, plays a critical role in the pathogenesis of kidney diseases, including acute kidney injury (AKI). However, reliable research methods for visualizing renal macrophages remain limited. In this study, we utilized positron emission tomography (PET) imaging combined with a radiolabeled ligand targeting colony-stimulating factor 1 receptor (CSF1R) to assess the potential of CSF1R-PET as a non-invasive tool for examining renal inflammation.

This study aimed to investigate the regulation of allergic rhinitis (AR) by methyltransferase 3 through an m6A-dependent mechanism, providing a theoretical foundation for its treatment.

Baicalein (Bai) has been found to alleviate the progression of tuberculosis (TB) by inhibiting mycobacterium tuberculosis (M.tb)-induced macrophage pyroptosis, so it may be used as an adjuvant treatment for TB. However, the underlying molecular mechanism of Bai remains unclear.

To summarize the current understanding of the pathogenesis of cerebral atherosclerosis to provide a basis for developing effective treatments and exploring future research directions for clinical application.

Heme oxygenase-1 (HO1, Hmox1) degrades excess heme and is considered an anti-oxidative and anti-inflammatory enzyme. Our previous studies in Hmox1 knockout mice revealed the induction of interferon-stimulated genes (ISGs) in all cell types analyzed, despite unchanged interferon production. Here, we sought to determine whether this induction is driven by intrinsic cellular mechanisms or extrinsic cues at the organismal level, and to identify the pathway underlying HO1-dependent ISG regulation. To this end, we analyzed how ISG expression changes in cultured cells exposed to stressors typical of Hmox1 knockout mice. Using murine wild-type and Hmox1-deficient (Hmox1 KO) fibroblasts, we found that under control conditions, the expression of most tested ISGs was independent of cellular HO1 status. We next examined the effects of extrinsic stressors, including hemolytic, oxidative, genotoxic, and replication stress, proinflammatory TNFα, and endogenous heme overload. TNFα, which is upregulated in Hmox1 knockout mice, was the sole and universal inducer of ISGs in both wild-type and Hmox1 KO fibroblasts. Unexpectedly, the response of Hmox1 KO cells to exogenous TNFα was weakened, likely due to impaired NF-κB activity and reduced nuclear retention of the p65 subunit. A similar decrease we observed for STAT1. Additionally, the presence of the TREX1 exonuclease in the nucleus pointed to compromised nuclear envelope integrity in HO-deficient cells. Notably, HO1 colocalizes with PARP1, a protein involved in envelope maintenance and regulation of cytoplasmic-nuclear transport. Inhibition of PARP1 with olaparib dampened TNFα-induced nuclear accumulation of p65 and STAT1 in wild-type cells, but not in Hmox1 KO counterparts. In summary, the inflammation observed in Hmox1-deficient mice appears to be the main cell-extrinsic driver of ISG induction in vivo. Despite this, the inflammatory response to exogenous TNFα is intrinsically attenuated in Hmox1 KO cells, likely due to decreased nuclear retention of NF-κB and STAT1.

1-carbon (1 C) metabolism is a key process required for nucleotide synthesis in activated lymphocytes and other immune-regulatory mechanisms. Commonly used and well-established drugs for the treatment of inflammatory disorders such as methotrexate and fluorouracil (5-FU) are known to interact with 1 C metabolism And provide appreciable clinical benefit despite challenges with tolerance And patient compliance. Advances in our understanding of the enzymes involved in 1 C metabolism coupled with differential expression highlights specific targeting of key enzymes may greatly enhance clinical efficacy And reduce side effect burden in comparison to the established therapies that are more indiscriminate in their mode of action. In recent years the methylenetetrahydrofolate dehydrogenase enzymes have gained increasing interest because of the role they play in 1 C metabolism in the immune system and in progression of inflammatory diseases. High expression of these enzymes in activated immune cells and inflamed tissues coupled with chemical tractability provides compelling reasons to actively consider these enzymes as novel therapeutic targets.

Traumatic brain injury (TBI) triggers neuroinflammation both acutely and chronically, the latter which might be involved in neurodegenerative disorders. Resolvins are neuroinflammatory modulators, hypothesized to improve resolution of inflammation. This study sought to explore sustained immunological responses after delayed treatment with resolvins utilizing novel tools for automated cellular assessments.

Natural killer (NK) cells are responsible for monitoring and eliminating malignant or virus-infected cells. To become activated, NK cells must upregulate oxidative phosphorylation and glycolysis to meet the high energetic demands associated with cytotoxic and effector functions. While glutamine can also fuel the tricarboxylic acid cycle through its conversion to alpha-ketoglutarate, the precise role of this pathway in NK-cell cytotoxic activity is unclear.

Alcohol consumption is the main cause of acute pancreatitis (AP). Alcohol combined with cerulein (CI) exerts direct damage to pancreas, with its mechanism undefined.

Immune cells can rewire their metabolism in response to various stimuli. Crosstalk between the nucleus and mitochondria allows for tight regulation of this metabolic reprogramming. Research has emerged showing several TCA cycle-derived metabolites exhibiting moonlighting functions in the nucleus, modulating chromatin modifications in order to control inflammation. These TCA cycle-derived metabolites include acetyl-CoA, α-ketoglutarate, succinate, fumarate, itaconate, and succinyl-CoA which can modify DNA or histone to drive or inhibit gene expression. In this review, we look at the mechanisms of TCA cycle metabolites' non-canonical functions in the nucleus in the context of inflammation. In addition, we discuss the known and possible links between these metabolites' nuclear moonlighting functions and the pathogenesis of diseases, including inflammatory diseases and cancers.

Aberrant activation of mTORC1 is clearly defined in TSC and causes uncontrolled cell growth. While mTORC1 inhibitors show efficacy in stabilising tumour growth in patients with TSC, they are not fully curative. Disease facets of TSC that are not restored with mTOR inhibitors might involve NF-κB. This study aimed to characterise NF-κB in the context of TSC.

Tumor necrosis factor-alpha (TNF) is an inflammatory cytokine implicated in the development of many chronic inflammatory diseases and TNF-α inhibitors (TNF-I) are frequently prescribed as treatment. Their malignancy risk is debated, with pro-oncogenic effects of decreased immune surveillance and anti-oncogenic effects of decreasing chronic inflammation. As such, the literature is inconclusive in the malignancy risk of these medications. Here we investigate the malignancy risk in patients with chronic TNF-I exposure.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the progressive loss of motor neurons. Although it is traditionally viewed as a neuron-centric disease, neurodegeneration is increasingly linked to immunosenescence and age-related immune dysfunction, but the mechanisms connecting immune ageing to neurodegeneration remain poorly understood. In this review, we explore how metabolic reprogramming, especially the loss of metabolic plasticity in senescent immune cells, drives neuroinflammation in ALS. Senescent immune cells, including microglia and T cells, exhibit mitochondrial dysfunction, redox imbalance, impaired autophagy, and altered nutrient-sensing pathways that impair their homeostatic and reparative capacities. These cells adopt a metabolically demanding pro-inflammatory phenotype, sustaining an inflammatory secretome while promoting glial activation and neuronal damage. Finally, we discuss how targeting immunometabolic pathways may offer new therapeutic opportunities to restore immune balance, mitigate neuroinflammation, and potentially slow ALS progression. Understanding the metabolic basis of immune ageing is essential for developing effective, age-tailored interventions for ALS.

Neutrophil extracellular traps (NETs) have been demonstrated to play an important role in acute myocardial infarction (AMI), but their specific mechanisms in the development of AMI have not been investigated.