bims-cediti 2026-05-03 papers

bims-cediti

Biomed News

on Cell death in innate immunity, inflammation, and tissue repair

Issue of 2026–05–03
28 papers selected by
Kateryna Shkarina, Universität Bonn

Human non-canonical inflammasomes activate CASP3 to limit intracellular Salmonella replication in macrophages.
Measurement and quantification of macrophage lytic cell death by kinetic microscopy.
Lipid-NLRP3 interplay in inflammasome regulation.
d-amino acids restrain macrophage IL-1β release through gasdermin D acetylation.
The dynamic and heterogeneous structure of the non-canonical inflammasome.
Cytosolic CTH senses bacterial lipoproteins and drives noncanonical inflammasome activation.
MTCH2 promotes BAX and BAK self-assembly and apoptotic pore growth.
Functional assessment and quantitative kinetic analysis of BAX activation using large unilamellar vesicles.
Site-Specific Photo-Cross-Linking Reveals CCT2 as a Regulator of cGAS-STING Signaling via Clearance of cGAS-DNA Condensates.
PARP4 ADP-ribosylates PIDD1 to complete a phospho/SUMO/PAR-ylation cascade that orchestrates PIDDosome assembly.
Glue it together: self-assembly of the antibacterial GTPase GBP1.
Semi-automated, quantitative immunofluorescence analysis of single cell cytochrome c release during apoptotic cell death.
Protective versus deleterious roles of pyroptosis in Mycobacterium avium and Mycobacterium marinum infections.
Regulated cell death in sepsis: reframing NETosis within the spectrum of apoptosis and inflammatory lytic death.
Cytoplasmic DNA sensors and their regulators.
Sorcin couples Annexin A11 recruitment and ESCRT-III assembly during plasma membrane repair.
NOD2 drives regenerative fetal-like reprogramming in the intestinal epithelium.
Modular double-stranded DNA recognition defines specificity of the exonuclease TREX1 in cGAS-STING control.
Innate lymphoid cells integrate sensing and plasticity to control fungal infections.
Lysosomal permeabilization by Group A Streptococcus releases proteins into the macrophage cytosol.
Submicrometre sampling of living cells by macrophages.
Stabilizing MARCH7 as a ferro-guardian against ferroptosis.
Identification of a novel MEFV L560F variant in a patient with Mollaret meningitis.
Context-dependent release of HMGB1: cell death mode, cell type and LPS stress drive monomer and heterocomplex formation.
Purification and analysis of cell death complexes.
Guarding versus self-guarding in innate immunity.
Glycolytic reprogramming and immune responses in macrophages: a crosstalk driven by bacterial infection.
Techniques for monitoring and isolating lipid droplets in cell death.

PLoS Pathog. 2026 Apr 27. 22(4): e1014178

Human non-canonical inflammasomes activate CASP3 to limit intracellular Salmonella replication in macrophages.

Madhura Kulkarni, Christopher M Bourne, Ashutosh B Mahale, Patrick M Exconde, Cecelia Murphy, Haley T Goodrow, Sofia Cervantes, Matilda Kardhashi, Mirai Kambayashi, William Yoo, Tristan J Wrong, Robert C Patio, Bohdana M Discher, Cornelius Y Taabazuing.

Inflammasomes are signaling platforms that activate inflammatory caspases to initiate innate immune responses. Canonical inflammasomes sense diverse threats and activate CASP1, which cleaves the pro-inflammatory cytokines IL-1β and IL-18 and the pore-forming protein gasdermin D (GSDMD) to induce pyroptosis. In contrast, the non-canonical inflammasome senses bacterial lipopolysaccharide (LPS) through CASP4 and CASP5 to induce pyroptosis. While CASP1 substrates are well defined, those of CASP4 and CASP5 remain less understood. Here, we show that intracellular LPS and the gram-negative bacterial pathogen Salmonella activate CASP4/5 in macrophages to directly cleave and activate CASP3 and CASP7. Activated CASP3 subsequently cleaves gasdermin E (GSDME). Surprisingly, CASP3, but not GSDME, was required for restricting intracellular Salmonella replication, suggesting a protective role for apoptotic signaling. We further find that most GSDMD cleavage during non-canonical signaling is mediated by CASP1. Consistent with this, loss of GSDMD, but not GSDME, reduced LDH release, establishing GSDMD as the primary driver of pyroptosis during LPS transfection. In contrast, during Salmonella infection, cell lysis occurred independently of both GSDMD and GSDME, suggesting the involvement of alternative lytic mechanisms. Finally, we demonstrate that CASP4/5 activation of CASP3/7 and GSDME occurs in human primary macrophages, defining CASP4/5 as dual apoptotic initiator and inflammatory caspases in innate immunity.

DOI: https://doi.org/10.1371/journal.ppat.1014178
Methods Cell Biol. 2026 ;pii: S0091-679X(26)00087-7. [Epub ahead of print]206 109-122

Measurement and quantification of macrophage lytic cell death by kinetic microscopy.

Sara Cahill, Fiachra Humphries.

  Macrophages are innate immune cells that are critical in the maintenance of tissue homeostasis and defense against pathogens. Programmed cell death is a critical tool macrophages use to clear pathogens and to alert surrounding cells to damage following induction of cell death. Diverse forms of cell death, including pyroptosis, necroptosis, ferroptosis, and secondary apoptosis following apoptosis, result in loss of plasma membrane integrity. Membrane disruption occurs following the oligomerization of pore-forming proteins into the cell membrane, releasing cytokines and damage associated molecular patterns that trigger the immune response. Thus, quantification of cell membrane permeability is an effective method for assessing cell death in macrophages. Many different factors, including macrophage polarization, stimulation, and inflammatory state, can impact macrophage predisposition to, and rate of, programmed cell death. Thus, it is important to have a method of accurately assessing macrophage cell death kinetically, rather than at a single endpoint. In this protocol, we describe a protocol for assessing cell death in macrophages by quantifying cell membrane permeability using kinetic microscopy. This method overcomes limitations of common single time point metrics for assessing cell death and is adaptable and scalable for use in assessing cell death across different cell types and treatment conditions.

Keywords:  Kinetic microscopy; Lytic; Macrophage; Pyroptosis

DOI:  https://doi.org/10.1016/bs.mcb.2026.02.019
Adv Biol Regul. 2026 Apr 20. pii: S2212-4926(26)00018-7. [Epub ahead of print]100 101161

Lipid-NLRP3 interplay in inflammasome regulation.

Maria Antonietta De Matteis, Antonello Prodomo, Giuliana Sarno.

  NLRP3 is a cytosolic innate immune sensor that detects PAMPs and DAMPs and, together with ASC, activates caspase-1 to drive IL-1β/IL-18 release and pyroptotic cell death. A major open question is how such diverse triggers funnel into a single molecular switch within NLRP3. Emerging evidence points to lipids as central integrators: they act as direct NLRP3 ligands, post-translational modifiers, and membrane scaffolds that choreograph inflammasome priming and assembly. Palmitoylation dynamically tunes NLRP3 stability, localization, and activation thresholds, while cardiolipin and PI4P function as organelle-specific lipid cues that recruit and activate NLRP3 at mitochondria and Golgi/endosomal membranes. NLRP3 also senses shifts in cholesterol, fatty acids, and ceramides, mechanistically linking lipid imbalance to cardiometabolic and inflammatory disease. In this review, we spotlight how specific lipid-NLRP3 interactions and lipid-driven post-translational modifications orchestrate inflammasome priming and activation across cellular membranes.

Keywords:  Inflammation-NLRP3; Lipids; Membranes; Organelle; Phosphoinositides

DOI:  https://doi.org/10.1016/j.jbior.2026.101161
Sci Adv. 2026 May;12(18): eaed1676

d-amino acids restrain macrophage IL-1β release through gasdermin D acetylation.

Zebiao Wu, Qilin Hu, Yinhao Shen, Jian Fu, Bingnan Liu, Meimei Zhang, Ifen Hung, Chunxue Liu, Wenkai Ren.

d-amino acids have been detected in various tissues; however, whether d-amino acids shape immune cell (e.g., macrophages) function remains undefined. Here, we demonstrated that inflammatory macrophages decrease mRNA expression of d-amino acid oxidase (DAAO) and d-aspartate oxidase (DDO) through nuclear factor κB (NF-κB) signaling. Notably, inhibition of DAAO or DDO increases the concentration of intracellular d-amino acids, consequently suppressing IL-1β release. Mechanistically, d-amino acids inhibit the formation of gasdermin D (GSDMD) oligomer via GSDMD-K146 acetylation. d-amino acids directly bind and increase the enzyme activity of mitochondrial pyruvate dehydrogenase (PDH), resulting in acetyl-coenzyme A production for acetylation. Consistently, d-Ala/d-Glu supplementation or myeloid-specific deletion of DDO attenuates lipopolysaccharides (LPS)-induced sepsis in mice. Collectively, our study reveals a mechanism involving acetylation mediated by d-amino acids in regulation of macrophage function, providing a potential therapeutic strategy for treating macrophage-associated inflammatory diseases.

DOI: https://doi.org/10.1126/sciadv.aed1676
bioRxiv. 2026 Apr 16. pii: 2026.04.15.718739. [Epub ahead of print]

The dynamic and heterogeneous structure of the non-canonical inflammasome.

Alexander I M Sever, James M Aramini, Jeffrey P Bonin, Huaying Zhao, Hanlin Wang, John L Rubinstein, Peter Schuck, Lewis E Kay.

Inflammasomes are high molecular weight complexes that play an integral role in the innate immune system, triggering an inflammatory cascade to protect against cellular stresses such as pathogenic bacteria. Both canonical and non-canonical inflammasomes have been described in the literature and detailed structural studies of many components of the more complex and larger canonical versions have been reported. In contrast, corresponding structures of the non-canonical inflammasome have not emerged even though it consists of only two components: lipopolysaccharide (LPS) from gram-negative bacteria, and one of caspase-4 or caspase-5 in humans or caspase-11 in mice. Here we determine the stoichiometry of the non-canonical inflammasome using size-exclusion chromatography coupled with UV, refractive index, and light-scattering measurements, showing that the non-canonical inflammasome is heterogeneous, comprised of three major complexes with different numbers of LPS and caspase molecules. Solution Nuclear Magnetic Resonance (NMR) spectroscopy studies of the N-terminal Caspase Activation and Recruitment Domain (CARD) of caspase-11, that binds LPS, show that it is largely unstructured in the absence of lipid, with pervasive dynamics on the μs-ms timescale. Formation of this complex increases the alpha-helical content of the CARD but the dynamics persist, multiple conformers are formed, and tertiary contacts are transient, consistent with formation of a molten globule. Our NMR results establish that the protease domain of caspase-11 is monomeric in isolation. As proteolysis is linked with dimerization, the protease domains are inactive in this state, but upon formation of the non-canonical inflammasome dimerization occurs, priming the complex for rapid processing of substrates.
Significance Statement: Animals respond to injury, infection, or toxic materials via a process called inflammation. At the molecular level inflammation involves formation of large machines - inflammasomes - that are instrumental in triggering cascades that lead to an immune response. Although structural studies of canonical inflammasomes have emerged, much less is known about the structures of non-canonical inflammasomes. Using solution Nuclear Magnetic Resonance (NMR) spectroscopy in concert with other biophysical approaches we show that non-canonical inflammasomes are highly dynamic and structurally heterogeneous, and we characterize the different sized inflammasome particles that are formed in terms of their composition. We also show that once formed, non-canonical inflammasomes are primed to rapidly cleave substrate molecules, necessary for propagating the immune response.

DOI: https://doi.org/10.64898/2026.04.15.718739
Nat Immunol. 2026 Apr 30.

Cytosolic CTH senses bacterial lipoproteins and drives noncanonical inflammasome activation.

Qiannv Liu, Chunlei Wang, Mengqian Li, Chun Kong, Zijian Zhong, Xiangyun Cheng, Xiangyang Geng, Zhixia Li, Chongyao Zhong, Dong Jiang, Xiuli Sun, Shuo Wang, Pengyan Xia.

Pathogen-associated molecules can have both membrane-associated and intracellular receptors. Bacterial lipoproteins are recognized by Toll-like receptor 2, but it is unclear whether they can also be sensed by cytoplasmic receptors. Here we found that bacterial lipoproteins could be recognized in the cytoplasm of macrophages by cystathionine γ-lyase (CTH) and hydrolyzed into lipid chains containing sulfhydryl groups. The hydrolyzed lipid chains form molecules containing four acylated chains linked through disulfide bonds, which further cleave caspase-11 and activate the noncanonical inflammasome. Changing the redox environment in macrophages affects their recognition of bacterial lipoproteins. CTH-deficient primary and immortalized macrophages do not trigger activation of the noncanonical inflammasome in the presence of intracellular bacterial lipoproteins, while CTH-deficient mice exhibit attenuated immune responses to infection with Staphylococcus aureus and Listeria monocytogenes. Our findings elucidate the molecular mechanisms by which macrophages recognize intracellular bacterial lipoproteins, as well as the regulatory relationship between cellular redox levels and infection resistance.

DOI: https://doi.org/10.1038/s41590-026-02511-9
Nat Struct Mol Biol. 2026 Apr 29.

MTCH2 promotes BAX and BAK self-assembly and apoptotic pore growth.

Hector Flores-Romero, Aida Pena-Blanco, Jonas Aufdermauer, Shashank Dadsena, Philip Neubert, Lisa Hohorst, Eileen Cors, Cristiana Zollo, Alvaro Larrañaga-SanMiguel, Jone Zaldunbide, Maria Nenchova, Yasmin Carvalho-Schaefer, Thomas Langer, Georg Häcker, Ana J Garcia-Saez.

During apoptosis, the BCL-2 family members BAX and BAK oligomerize and form a pore to mediate the decisive step of mitochondrial outer membrane permeabilization. However, the contribution of additional cellular components to apoptotic pore dynamics remains poorly understood. Here we map the protein environment of the apoptotic pore using in situ proximity labeling and identify the mitochondrial carrier homolog protein MTCH2 localizing nearby BAX and BAK assemblies specifically under apoptotic conditions. We show that cells lacking MTCH2 exhibit delayed BAX and BAK oligomerization at the single-particle level, which can be rescued by addition of lysophosphatidic acid. Accordingly, MTCH2 depletion decreases not only apoptosis sensitivity but also sublethal mitochondrial permeabilization during bacterial infection, mitochondrial DNA release into the cytosol and cGAS-STING activation under impaired caspases. Our findings uncover a key role of MTCH2 in promoting BAX and BAK high-order assembly with functional consequences for apoptotic pore growth and downstream responses.

DOI: https://doi.org/10.1038/s41594-026-01805-8
Methods Cell Biol. 2026 ;pii: S0091-679X(26)00085-3. [Epub ahead of print]206 55-80

Functional assessment and quantitative kinetic analysis of BAX activation using large unilamellar vesicles.

Jesse D Gelles, Thedoe Nyunt, Md Abdullah Al Noman, Jerry Edward Chipuk.

  A myriad of diverse developmental and pro-death signals converge on the mitochondrial pathway of apoptosis, which is governed by the BCL‑2 family of proteins. Comprised of both pro- and anti-apoptotic family members, the BCL‑2 family functions to regulate mitochondrial outer membrane permeabilization (MOMP), often considered the "point of no return" in which a cell commits to an apoptotic outcome. Specifically, the effector BCL‑2 family proteins, BAX and BAK, are responsible for inducing MOMP and therefore investigations into their structural, cellular, and pharmacological regulation are critical to understanding the cellular commitment to apoptosis. A gold standard methodology for studying activation of BAX or BAK is the permeabilization of large unilamellar vesicles (LUVs), which are biochemically-defined model liposomes that mimic the major lipid composition of the outer mitochondrial membrane (OMM). Here, we provide a detailed protocol for generating LUVs containing a fluorescent dye/quencher pair to monitor real-time BAX activation and membrane permeabilization using a standard plate reader. Additionally, we detail example assay strategies to model interactions within the BCL‑2 family and provide a robust mathematical model for fitting and parameterizing kinetic LUV permeabilization data.

Keywords:  Apoptosis; BAX; BCL‑2 family; Large unilamellar vesicles; MOMP; Regulated dell death

DOI:  https://doi.org/10.1016/bs.mcb.2026.02.017
ACS Chem Biol. 2026 Apr 27.

Site-Specific Photo-Cross-Linking Reveals CCT2 as a Regulator of cGAS-STING Signaling via Clearance of cGAS-DNA Condensates.

Xinyu Zhang, Yi Zhou, Hang Yin.

Cyclic GMP-AMP synthase (cGAS) serves as a cytosolic DNA sensor that detects double-stranded DNA (dsDNA) and responds by producing 2'3'-cyclic GMP-AMP (cGAMP), which in turn initiates downstream signaling events that drive innate immune activation. Tight control of cGAS activity is required to preserve immune balance; however, the molecular factors that fine-tune its activation dynamics and protein stability are not yet fully understood. Here, we employed a residue-resolved photo-cross-linking approach coupled with quantitative proteomics to profile cGAS regulators, which uncovered the chaperonin TRiC subunit CCT2 as a previously unappreciated cGAS-associated factor. Functional analyses demonstrated that CCT2 attenuates cGAS-STING signaling by facilitating autophagy-mediated turnover of DNA-bound cGAS aggregates, thereby limiting the persistence of immunostimulatory cytosolic DNA signals and ensuring appropriate immune responses. Collectively, these findings demonstrate that site-specific protein photo-cross-linking provides a powerful means to interrogate protein-protein interactions and define CCT2 as a key negative modulator of cGAS, with implications for therapeutic modulation of antiviral immunity.

DOI: https://doi.org/10.1021/acschembio.6c00142
Sci Adv. 2026 May;12(18): eadz9284

PARP4 ADP-ribosylates PIDD1 to complete a phospho/SUMO/PAR-ylation cascade that orchestrates PIDDosome assembly.

Richa B Shah, Yuanyuan Li, Ashley Person, Léonie Frigon, Nina Ðukić, Ela Kini, Raymond Chen, John M Pascal, Michael S Cohen, Ivan Ahel, John D Haley, Samuel Sidi.

Adenosine diphosphate (ADP) ribosylation (ADPr) regulates multiple stress responses, yet substrates in the apoptotic machinery remain elusive. We show that a single, DNA damage-induced ADPr event controls proapoptotic PIDDosome (PIDD1/RAIDD/caspase-2) formation in response to unresolved interstrand DNA cross-links (ICL). ADPr targets conserved E783 in the PIDD1 death domain (DD); is catalyzed by poly(ADP-ribose) polymerase 4 (PARP4), a phylogenetically orphan PARP of previously unknown function; is reversed by the ribosylhydrolase activity of PARP14; and is triggered by Ataxia Telangiectasia and RAD3-related (ATR) phosphorylation-induced, PIAS1-mediated SUMOylation of the PIDD1 DD, which enables PARP4 docking. PIDD1 ADPr is dispensable for the recruitments of RAIDD and caspase-2 but essential for the dimerization of the caspase. Hence, denying E783 ADPr spares the onset of PIDDosome assembly but blocks its completion, thus eliminating caspase-2 activation and ensuing apoptosis. Conversely, removal of PARP14 forces apoptosis, even in cells with tolerable damage. The data identify PARP4 as an ICL response effector and illuminate a three-step modification sequence of the PIDD1 DD that conducts PIDDosome assembly from initiation to completion.

DOI: https://doi.org/10.1126/sciadv.adz9284
Biol Chem. 2026 Apr 21.

Glue it together: self-assembly of the antibacterial GTPase GBP1.

Miriam Kutsch, Christian Herrmann.

  Guanylate-binding proteins (GBPs) are interferon-inducible large GTPases that play a central role in cell-autonomous immunity against intracellular bacterial pathogens. A defining feature of GBPs is their ability to translate GTP binding and hydrolysis into large-scale conformational rearrangements that drive self-assembly into higher-order structures, including dimers, polymers, and membrane-associated coatomers. This review integrates insights from biochemical, biophysical, structural, and cell biology studies to summarize current mechanistic models of GBP self-assembly, with a particular focus on human GBP1. We highlight how GTP hydrolysis-driven GBP1 polymers and coatomers act as self-regulating nanomachineries that recognize and remodel the pathogen-associated molecular pattern lipopolysaccharide, thereby fulfilling a dual function as immune sensor and effector in non-canonical inflammasome activation and bacterial membrane disruption. By directly linking nucleotide binding kinetics, enzymatic activity, and assembly dynamics to cellular and infection-related phenotypes, this review places decades of biochemical and biophysical work on GBP1 into a clear physiological context of antibacterial host defense.

Keywords:  GMP production; bacterial outer membrane; dimer building block; host-pathogen interaction; lipopolysaccharide sensor; open conformation

DOI:  https://doi.org/10.1515/hsz-2026-0112
Methods Cell Biol. 2026 ;pii: S0091-679X(26)00095-6. [Epub ahead of print]206 23-42

Semi-automated, quantitative immunofluorescence analysis of single cell cytochrome c release during apoptotic cell death.

Fabian Klötzer, Daniela Stöhr, Markus Rehm.

  Apoptosis, a tightly regulated form of programmed cell death, eliminates damaged or malignant cells and is triggered by internal or external stress signals. A critical decision point is mitochondrial outer membrane permeabilization (MOMP), governed by BCL-2 family proteins. Pro-apoptotic members such as BAX and BAK form pores in the mitochondrial outer membrane, releasing intermembrane space proteins like cytochrome c into the cytoplasm. Once cytosolic, cytochrome c binds APAF-1 to form the apoptosome, which activates caspase-9 and subsequently caspase-3, driving apoptosis through cleavage of key cellular substrates. Cytochrome c release serves as a hallmark and point of no return in the apoptotic cascade. However, cytochrome c release can be variable, occurring at submaximal levels or from only a subset of mitochondria, which complicates detection in heterogeneous cell populations. To address this, we developed a semi-automated imaging-based method to quantify cytochrome c release at the single-cell level using immunofluorescence microscopy. Our approach uses CellProfiler, an open-source image analysis platform, to implement a pipeline that segments adherent cells into nuclear, mitochondrial, and cytoplasmic compartments based on compartment-specific reference stains. The pipeline quantifies cytochrome c distribution across these compartments, calculating the ratio of mitochondrially retained to cytoplasmic cytochrome c for each cell. Automation of segmentation and measurement ensures rapid, robust, and reproducible analysis, with only image acquisition and data interpretation performed manually. This method provides a quantitative readout of MOMP and can be readily adapted to any immunofluorescence-detectable protein given an appropriate compartmental marker, expanding its utility for broader cellular studies.

Keywords:  Apoptosis; Cell segmentation; CellProfiler; Cytochrome c release; Immunofluorescene staining; MOMP; Semi-automated quantification

DOI:  https://doi.org/10.1016/bs.mcb.2026.03.002
Cell Death Dis. 2026 Apr 30.

Protective versus deleterious roles of pyroptosis in Mycobacterium avium and Mycobacterium marinum infections.

Wanbin Hu, Federico Pagliaro, Herman P Spaink, Annemarie H Meijer.

Mycobacterium avium, a slow-growing nontuberculous mycobacterium (NTM), is the main cause of life-threatening NTM infections, which are globally on the rise. Unlike Mycobacterium tuberculosis and Mycobacterium marinum, M. avium lacks ESX-1, a subtype of Type VII secretion system (T7SS) and a key virulence determinant. The absence of ESX-1 in M. avium raises questions about its alternative intracellular survival strategies. To investigate M. avium pathogenesis, we exploited our recently established infection model in zebrafish larvae, enabling live imaging of early host-pathogen interactions. Macrophage depletion significantly increased M. avium burden and larval mortality, while neutrophil depletion had no major effect, emphasizing macrophages as key defenders against M. avium. In support, imaging of tnfa activation showed that macrophages polarized to a proinflammatory phenotype. However, like M. marinum, M. avium exploits chemokine receptor Cxcr3.2 signaling in macrophages for its expansion in granuloma-like clusters. Both M. avium and M. marinum preferentially infected macrophages, but M. avium-induced granuloma-like clusters were more compact and exhibited less cell death. Supporting this, lytic cell death pathways were enriched in M. marinum but not M. avium transcriptome signatures. Consequently, we investigated pyroptosis, an important form of inflammation-induced lytic cell death. We found that knockdown of critical mediators of pyroptosis, namely inflammatory caspase a (caspa) and gasdermin Eb (gsdmeb), produced opposing effects on the two mycobacterial pathogens, indicating a host-protective role during M. avium infection, while exacerbating M. marinum growth. These findings highlight the interaction with host cell death signaling as a determining factor for the pathogenic potential of mycobacterial species.

DOI: https://doi.org/10.1038/s41419-026-08810-1
Inflamm Res. 2026 Apr 28. pii: 106. [Epub ahead of print]75(1):

Regulated cell death in sepsis: reframing NETosis within the spectrum of apoptosis and inflammatory lytic death.

Toshiaki Iba, Hideshi Okada, Isao Nagaoka, Ricard Ferrer, Jerrold H Levy.

   BACKGROUND: Sepsis is characterized by dysregulated inflammation leading to organ dysfunction. While immune activation and metabolic stress are central features, accumulating evidence suggests that regulated cell death programs actively influence inflammatory trajectories rather than serving as passive end-stage events. Apoptosis, pyroptosis, necroptosis, ferroptosis, and neutrophil extracellular trap (NET) formation have each been implicated in sepsis; however, their relative hierarchy, temporal dynamics, and compartment-specific relevance remain incompletely defined.
OBJECTIVE: To synthesize current evidence on regulated cell death pathways in sepsis and to propose a phase-specific and compartment-oriented framework that integrates apoptotic, inflammatory lytic, and NET-associated mechanisms within a unified inflammatory model.
METHODS: A narrative review of experimental, translational, and clinical studies examining apoptosis, pyroptosis, necroptosis, ferroptosis, PANoptosis, and NETosis in sepsis and related inflammatory states was conducted. Emphasis was placed on signaling dependency, inflammatory consequences, temporal phase distinctions, and cellular compartment heterogeneity.
RESULTS: Apoptosis remains the dominant leukocyte death program associated with late-phase immune depletion and immunosuppression. In contrast, inflammasome-mediated pyroptosis and RIPK1/RIPK3-dependent necroptosis amplify early hyperinflammatory responses by inducing membrane permeabilization and damage-associated molecular pattern release. Ferroptosis represents an emerging iron-dependent metabolic-inflammatory interface with potential organ-specific relevance, although clinical validation remains limited. NET formation, often interpreted as a distinct death program, is more appropriately understood as a context-dependent effector mechanism linking innate immunity to thromboinflammation rather than representing the predominant terminal fate of leukocytes in sepsis. Increasing evidence supports pathway crosstalk and PANoptotic integration, suggesting that regulated cell death programs function as overlapping inflammatory networks rather than isolated processes.

Keywords:  Immunothrombus; Inflammation; Leukocyte; Neutrophil extracellular trap; Programmed cell death

DOI:  https://doi.org/10.1007/s00011-026-02261-2
Biomed J. 2026 Apr 28. pii: S2319-4170(26)00044-2. [Epub ahead of print] 100988

Cytoplasmic DNA sensors and their regulators.

Roger J Eloiflin, Isabelle K Vila, Nadine Laguette.

Deoxyribonucleic acid (DNA) is naturally contained within the mitochondria and nucleus of cells. DNA localized outside of these areas is generally considered to constitute damage-associated molecular patterns (DAMP) that trigger innate immune responses. The variety of these immune-stimulatory nucleic acids, coupled with the variety of proteins known to interact with DNA DAMPS and to act as sensors or regulators, is an important layer of regulation that defines the nature and breadth of inflammatory responses within a given cell or tissue. In this review, we describe the various DNA substrates that co-exist in cells, how they are sensed, and what regulates their cytoplasmic availability and recognition. We subsequently discuss how this complexity may dictate tissue-specific immune response and suggest that a more integrated view of the interconnection between pathways is necessary in order to define molecular targets for treatment intervention in inflammatory pathologies.

DOI: https://doi.org/10.1016/j.bj.2026.100988
bioRxiv. 2026 Apr 13. pii: 2026.04.10.717788. [Epub ahead of print]

Sorcin couples Annexin A11 recruitment and ESCRT-III assembly during plasma membrane repair.

Jordan Matthew Ngo, Justin Krish Williams, Abinayaa Murugupandiyan, Randy Schekman.

The absence of a cell wall affords animal cells diverse functionality at the cost of acute sensitization to plasma membrane (PM) damage. Thus, animal cells tightly monitor and maintain the integrity of their PM to prevent cell death. Genetic loss of PM repair factors is associated with human diseases including muscular dystrophy and neurodegeneration. Despite evidence that annexin and endosomal sorting complex required for transport (ESCRT) proteins are required for PM repair, the extent to which their recruitment is coordinated at sites of membrane damage is unclear. Here, we identify sorcin as a new PM repair factor that directly couples annexin A11 (ANXA11)-mediated sensing of PM damage and ESCRT-III assembly. We demonstrate that ANXA11, recruited to the PM upon damage-induced calcium influx, serves as an anchor that facilitates the sequential recruitment of sorcin and ESCRT-III at PM lesions. Our data highlight mechanistic and topological similarities between the budding of membrane-enveloped viruses and damage-induced microvesicles. We propose that they share a common mechanism of membrane budding and speculate that membrane-enveloped viruses may have co-opted this host pathway of PM ESCRT recruitment to facilitate virion assembly and propagation.

DOI: https://doi.org/10.64898/2026.04.10.717788
Proc Natl Acad Sci U S A. 2026 May 05. 123(18): e2519981123

NOD2 drives regenerative fetal-like reprogramming in the intestinal epithelium.

Derek K L Tsang, Charles Maisonneuve, Arshad Ayyaz, Elisabeth G Foerster, Marry Nissan, Lauren Baerg, Daniel Trcka, Bibaswan Ghoshal, Boyan K Tsankov, Giuliano Bayer, Madison Denney, Catherine J Streutker, Jeffrey L Wrana, Stephen E Girardin, Dana J Philpott.

  Inflammatory injury to the intestine triggers a reprogramming of the intestinal epithelium to a fetal-like state that drives rapid restoration of the epithelial barrier. Although the intestinal microbiota is a key modulator of inflammation, its role in influencing epithelial fetal-like stem cell reprogramming and consequent restitution remains unclear. Using irradiation (IR) injury as a model for small intestinal epithelium injury and repair, we found that the intestinal microbiota accelerated epithelial restitution by amplifying a repair-associated inflammatory response that promoted the emergence of fetal-like intestinal epithelial cells (IECs), marked by Ly6a and Clu. NOD2, the strongest genetic link to the development of Crohn's disease, was found to be expressed in fetal-like IECs following injury. Employing an ileal organoid model, we demonstrated that NOD2 activation by its peptidoglycan ligand potentiated an inflammatory gene signature characterized by interferon signaling, concurrent with enterocyte recovery. NOD2 deficiency exacerbated epithelial apoptosis following IR injury, whereas epithelial-specific NOD2 signaling promoted fetal-like IEC emergence and increased epithelial proliferation. Collectively, these findings reveal a pivotal role for the microbiota and NOD2-mediated microbial sensing in regulating fetal-like IEC fate after injury, thus contributing to the protective function of this microbial sensor during intestinal inflammation.

Keywords:  NOD2; fetal-like reversion; inflammation; intestinal regeneration; microbiota

DOI:  https://doi.org/10.1073/pnas.2519981123
Immunity. 2026 Apr 24. pii: S1074-7613(26)00136-6. [Epub ahead of print]

Modular double-stranded DNA recognition defines specificity of the exonuclease TREX1 in cGAS-STING control.

Jiali Zhu, Lei Wang, Changjie Lin, Shuchen Bian, Xiaohan Luan, Yingying Sui, Luhang Piao, Wen Zhou.

  TREX1, but not other DEDDh exonucleases, restrains cyclic GMP-AMP synthase- stimulator of interferon genes (cGAS-STING) activation by cytosolic DNA. Here, we examined the mechanisms underlying this specificity. Biochemical comparison of TREX1 and TREX2 mapped TREX1's ability to degrade dsDNA and suppress cGAS signaling to a single amino acid, R128. Metazoan TREX proteins containing an R128-equivalent functioned as double-stranded DNA (dsDNA) nucleases, often alongside cGAS-like receptors, defining an evolutionarily conserved family. Structurally, TREX1-R128 contacted the non-substrate strand to enable efficient dsDNA binding and cleavage. Examination of conserved structural features of the R128-containing TREX family further identified a substrate-sensing loop that inserted into the active-site cleft and stabilized the substrate strand. In human TREX1, a distinct auxiliary DNA-binding surface (B-site) strengthened DNA engagement and cGAS restraint independent of DNase activity, suggesting a competitive function. Autoimmune-associated TREX1 mutations mapped to TREX1-dsDNA interfaces, and mutations disrupting these interactions increased cGAS-STING signaling and antitumor immunity in murine models. Thus, a modular dsDNA-recognition architecture defines TREX1 specificity and regulation of the cGAS-STING pathway.

Keywords:  DNA sensing; STING; TREX1; cGAS; evolutionary immunology; innate immunity; nuclease; structural biology; tumor immunity

DOI:  https://doi.org/10.1016/j.immuni.2026.03.025
Cell Rep. 2026 Apr 20. pii: S2211-1247(26)00218-4. [Epub ahead of print]45(4): 117140

Innate lymphoid cells integrate sensing and plasticity to control fungal infections.

Irina Tsymala, Philipp Penninger, Filomena Nogueira, Sabrina Jenull, Trinh Phan-Canh, Helena Brezovec, Magdalena Teufl, Sara Miranda, Wilfried Ellmeier, Nicole Boucheron, Birgit Strobl, Dagmar Gotthardt, Veronika Sexl, Karl Kuchler.

  Innate lymphoid cells (ILCs) are crucial regulators of tissue immunity. Here, we demonstrate that pulmonary ILCs can sense fungal components, leading to their activation. Mechanistically, we identify Syk/p38-dependent signaling as one of the key drivers of ILC activation following fungal challenges. Aspergillus fumigatus infection reshaped this response in vivo, creating a cytokine milieu that promoted ILC3 induction. We identified interleukin (IL)-23, IL-1β, and TGF-β as drivers of ILC2 conversion and IL-23 and IL-1β as triggers for ILC1s obtaining an ILC3 phenotype in vitro. Moreover, adoptive ILC transfer into Rag2-/-Il2rg-/- mice restrained excessive inflammation, while ILCs lacking the intracellular non-receptor tyrosine kinase Tec showed enhanced ILC1 proliferation and reduced fungal burden. Consequently, transfer of Tec-deficient ILCs leads to better survival by enhancing antifungal immunity. These findings uncover hitherto unrecognized roles for ILCs as early modulators of antifungal immunity. Hence, targeting Tec signaling in ILCs may offer a therapeutic strategy to enhance antifungal immunity.

Keywords:  Aspergillus; CP: immunology; CP: microbiology; Candida glabrata; Candida species; IL-17A; ILC plasticity; ILCs; Syk signaling; Tec kinase; antifungal immunity; aspergillosis; fumigatus; host defense; inflammation; innate immunity; innate lymphoid cells; mucosal immunity; pulmonary infection

DOI:  https://doi.org/10.1016/j.celrep.2026.117140
Infect Immun. 2026 Apr 30. e0019926

Lysosomal permeabilization by Group A Streptococcus releases proteins into the macrophage cytosol.

Ava Quezada, Kevin Lord, Cheldon Alcantara, Claire Delahunty, Kevin Kim, Olivia Okamoto, John R Yates, Cheryl Y M Okumura.

  The human-specific bacterial pathogen Group A Streptococcus (GAS) is a significant cause of morbidity and mortality due to its ability to cause severe invasive infection. Although macrophages are important for controlling GAS infection, we and others have demonstrated that GAS can persist in macrophages by perforating the phagolysosome using the pore-forming toxin streptolysin O (SLO). In this study, we examined how phagosomal perforation releases lysosomal and bacterial proteins into the cytosol and alters cytosolic protein content in the macrophage. Using IL-1β as a measure of intracellular pathogen detection, we confirmed that cytosolic preparations from macrophages infected with either wild-type (WT) or SLO-deficient (ΔSLO) bacteria contained new proteins that are absent in uninfected cytosol controls. Proteomic analysis revealed distinct cytosolic protein profiles in both WT- and ΔSLO-infected macrophages. M1 protein was detected only in the cytosol of WT-infected macrophages and corresponded with the IL-1β response, indicating SLO-mediated release of M1 protein from the phagosome, and providing a mechanism for cytosolic recognition of this virulence factor. Unexpectedly, cytosolic extracts of both WT- and ΔSLO-infected macrophages contained histone proteins H1-H4, suggesting that nucleosomal complexes are released into the cytosol during GAS infection. Our work reveals both a mechanism for the activation of the inflammatory response on a cellular level, and the surprising consequences of phagosomal perforation in GAS infections. These responses may collectively contribute to the pathologies observed during severe invasive GAS infection, and can help inform therapies aimed at improving macrophage function and patient outcomes.

Keywords:  Group A Streptococcus; IL-1β; histones; inflammasome; lysosome; macrophage; streptolysin O (SLO)

DOI:  https://doi.org/10.1128/iai.00199-26
Nature. 2026 Apr 29.

Submicrometre sampling of living cells by macrophages.

Amy C Fan, Rukman R Thota, Nina Serwas, Vivasvan S Vykunta, Kyle Marchuk, Megan K Ruhland, Lauren Liu, Grace Johnson, Austin Edwards, Matthew F Krummel.

An effective immune system must sample and develop healthy self-identity to prevent autoimmunity and to discern pathogenic insults1-3. Self-proteins are presented to T cells in the thymus during immune cell development2,3 and must be presented throughout the body to maintain regulatory T cell populations4-6 and to provide tonic signals to sustain conventional T cells over time7-9. Observations of continuous apoptosis in some organs together with the ingestion of that material by myeloid populations has led to a conventional understanding of ongoing cell death as a major source of self-antigens10. Here we used a series of companion imaging and vesicular labelling technologies to reveal an alternative process undertaken by macrophages that results in non-destructive, direct sampling of living cells. This process requires cell-cell contact, does not require caspase activation and occurs via trogocytosis-like stretching of the target cell into the macrophage, which leads to the generation of submicrometre-sized vesicles that contain cytoplasm. Using a high-dimensional flow-based method for labelling vesicles, we demonstrate that live-sampled material is distinctly processed and is poorly subjected to fusion with lysosomes. The material also produces differential effects on the presentation of antigen to CD4 T cells compared with CD8 T cells. Disruption of this trafficking by redirecting antigen to the lysosome significantly reduced the associated macrophage-mediated priming of CD8 T cells. These results demonstrate an important and substantial sampling of living cells by the immune system, with clear consequences for maintaining the border of immunity.

DOI: https://doi.org/10.1038/s41586-026-10435-5
Cell. 2026 Apr 27. pii: S0092-8674(26)00388-0. [Epub ahead of print]

Stabilizing MARCH7 as a ferro-guardian against ferroptosis.

Wenxiang Huang, Ruijun Wang, Xinquan Yang, Shuangjie Yang, Xueliang Yang, Gaolu He, Songjun Dai, Caizhi Li, Lianchao Gao, Tingting Zhang, Peng Zhang, Ruihan Chen, Keke Zheng, Junbing Wu, Junxia Min, Qian Ba, Fudi Wang, Qiang Zhang.

  Ferroptosis is an iron-dependent form of regulated cell death. However, the critical regulators that restrain iron overload to suppress ferroptosis remain undefined. Utilizing multi-omics, we identify the E3 ubiquitin ligase membrane-associated RING-CH 7 (MARCH7) as a non-redundant, dual suppressor of ferroptosis via direct regulation of intracellular iron homeostasis. Mechanistically, MARCH7 ubiquitylates nuclear receptor coactivator 4 (NCOA4) at residue Lys42 by K48-linked ubiquitination, promoting NCOA4 proteasomal degradation and reducing the labile iron pool. Concomitantly, MARCH7 modifies transferrin receptor 1 (TFR1) at residue Lys53 by K63 ubiquitination, restricting its plasma membrane translocation and thereby inhibiting cellular iron uptake. Through high-content screening, we further identify emodinanthrone (EmodAn) as a specific MARCH7 stabilizer with a strong cardioprotective effect in rodent models by blocking ferroptosis. In conclusion, our findings define an iron homeostasis regulatory hub for ferroptosis and suggest that stabilizing MARCH7 is a promising therapeutic strategy to protect against ferroptosis- or iron-overload-induced diseases.

Keywords:  MARCH7; NCOA4; TFR1; ferroptosis; iron overload; myocardial protection; ubiquitination

DOI:  https://doi.org/10.1016/j.cell.2026.03.052
Gene. 2026 Apr 28. pii: S0378-1119(26)00206-4. [Epub ahead of print] 150196

Identification of a novel MEFV L560F variant in a patient with Mollaret meningitis.

Hideo Handa, Akiyuki Uzawa, Dai Kishida, Yoshitaka Honda, Naoya Iwata, Hitoshi Kubosawa, Hiroki Kano, Yuki Nakagawa, Setsu Sawai, Atsuhiko Sugiyama, Satoshi Kuwabara.

   OBJECTIVES: Mollaret meningitis is a rare neurological condition characterized by recurrent episodes of aseptic meningitis and the presence of monocyte-derived atypical cells (Mollaret cells) in the cerebrospinal fluid. It is most frequently associated with herpes simplex virus type 2 infection; however, autoimmune and inflammatory disorders, including familial Mediterranean fever, have also been implicated in its pathogenesis. Here, we report an unreported MEFV variant in a patient with Mollaret meningitis and aim to evaluate its pathogenic relevance through in vitro functional analyses.
METHODS: MEFV exon sequencing was performed in a patient with Mollaret meningitis who showed a favorable response to colchicine treatment. Functional analyses were conducted using THP-1 cell lines. THP-1 monocytes were transfected with the identified MEFV variant or a wild-type MEFV construct, and their effects on cell death and apoptosis-associated speck-like protein containing a Caspase Recruitment Domain (ASC) speck formation were evaluated, with or without UCN-01 stimulation.
RESULTS: MEFV exon sequencing revealed a novel heterozygous L560F variant (c.1678C > T). Functional analyses demonstrated that, compared with the wild-type MEFV, the L560F variant induced more pronounced cell death and ASC speck formation following UCN-01 stimulation, supporting the pathogenic role of this novel variant.
DISCUSSION: The L560F variant represents a novel MEFV variant with functional evidence supporting pathogenicity. The clinical effectiveness of colchicine in this patient further supports the pathogenicity of the L560F variant.

Keywords:  Familial Mediterranean Fever; Inflammasome; MEFV gene; Mollaret meningitis; Pyrin

DOI:  https://doi.org/10.1016/j.gene.2026.150196
Mol Med. 2026 May 02.

Context-dependent release of HMGB1: cell death mode, cell type and LPS stress drive monomer and heterocomplex formation.

Harini Pechiappan, Rebecka Heinbäck, Sigrid Stråtveit Gravning, Kjerstin Jakobsen, Richard Davies, Cecilia Aulin, Helena Erlandsson Harris.

   BACKGROUND: HMGB1 acts as an alarmin when released from stressed or dying cells. In vitro, HMGB1 has previously been demonstrated to readily form complexes with other molecules and through intermolecular disulfide bond formation form homodimers. Recently, dimerized HMGB1 was identified in serum of LPS-challenged mice. In cancer, HMGB1 has been described as having both tumour-promoting and tumour-suppressing features, possibly dependent on the form of HMGB1 released into the tumour microenvironment. Factors determining the form in which HMGB1 is released remain, however, largely unexplored. We therefore investigated the form of HMGB1 released during different cell death modes and in response to LPS stress using various tumour cell lines.
METHODS: Supernatants were collected from ten non- and LPS-treated tumour cell lines and necrotic, apoptotic and pyroptotic THP-1 monocytic cells, to assess active and passive secretion of HMGB1. Released proteins were concentrated by TCA-precipitation and analysed by Western blotting under reducing and non-reducing conditions to detect monomeric, dimeric, or HMGB1-protein complexes. Co-immunoprecipitation and LC-MS/MS were used to identify binding partners of extracellular HMGB1.
RESULTS: Tumour cells were found to release monomeric HMGB1 and HMGB1 heterocomplexes in the 50-60 kDa range, as indicated by their persistent high molecular weight under reducing conditions. Furthermore, we identified that HMGB1 interacts with ribosomal proteins, histone H2B, and SRP9 following LPS treatment of microglial SIM-A9 cells.
CONCLUSIONS: HMGB1 readily formed heterocomplexes, but not homodimers in vitro across multiple cell lines, with differences between LPS-treated and untreated conditions. The form of released HMGB1 was influenced by cell type, cell death mode, and LPS stress.

Keywords:  HMGB1; alarmin; apoptosis; cancer; necrosis; pyroptosis

DOI:  https://doi.org/10.1186/s10020-026-01489-2
Methods Cell Biol. 2026 ;pii: S0091-679X(26)00086-5. [Epub ahead of print]206 43-54

Purification and analysis of cell death complexes.

Lingjie Yan, Daichao Xu.

  Cell death complexes, such as the complex II and necrosome, play pivotal roles in executing programmed cell death pathways. Understanding the composition and dynamics of these complexes is essential for elucidating the molecular mechanisms underlying apoptosis and necroptosis. This protocol describes a robust method for the affinity purification of endogenous or tagged cell death complexes from mammalian cells, followed by analysis through western blotting and optional mass spectrometry. This approach enables investigation of protein-protein interactions and the dynamic assembly of complexes under various stimuli or genetic conditions, providing insights into their mechanisms in health and disease.

Keywords:  Apoptosis; Complex; Necroptosis; Purification; TNF-α

DOI:  https://doi.org/10.1016/bs.mcb.2026.02.018
Evolution. 2026 Apr 25. pii: qpag075. [Epub ahead of print]

Guarding versus self-guarding in innate immunity.

Ben Ashby, Alyssa Anderson.

Hosts have evolved a variety of innate immune responses to pathogens. In many cases, hosts directly detect pathogen-associated molecular patterns (PAMPs) or pathogen effectors to trigger an immune response. However, hosts may also detect pathogens indirectly through 'guarding', whereby immune receptors ('guards') monitor the effects of pathogens (e.g., modification of target cells) rather than the pathogens themselves. Guarding poses a different evolutionary challenge for pathogens than direct recognition of PAMPs, as replication may necessitate the modification or disruption of guarded host proteins ('guardees'). Recently, self-guarding has been discovered, in which the host target functions as both guard and guardee. Self-guarding appears to present an intractable problem for pathogens: modification of the host target may benefit replication, but also triggers an immune response. If self-guarding creates an apparently inescapable detection mechanism, why has self-guarding only recently been discovered? Here, we use mathematical models of within-host pathogen and immune dynamics to compare guarding and self-guarding architectures. We show that self-guarding leads to a more rapid immune response and faster pathogen suppression, but is also more prone to false-positive immune responses, likely imposing greater costs through autoimmunity. We therefore hypothesise that the greater potential for false-positive immune responses may limit the conditions under which self-guarding evolves.

DOI: https://doi.org/10.1093/evolut/qpag075
Front Immunol. 2026 ;17 1783266

Glycolytic reprogramming and immune responses in macrophages: a crosstalk driven by bacterial infection.

Wei-Ren Wang, Lin Yan, Chuan-Ying Zhao, Cong-Cong He, Xing-Hua Gao.

  Macrophage glycolytic reprogramming during bacterial infection is a recognized metabolic shift with profound yet incompletely defined immunological consequences. This review delineates how this metabolic remodeling extends beyond energy provision to function as an integral immunoregulatory platform. We systematically examine the dual roles of key metabolic components, including the conformational dynamics of pyruvate kinase M2 that couple metabolic flux with inflammatory gene transcription, and the NAD+/NADH ratio that balances inflammasome activation against interferon responses. The review further explores how metabolites like lactate, succinate, and itaconate mediate immunomodulation through novel post-translational modifications, including histone lactylation and protein succinylation. Crucially, we analyze how diverse bacterial pathogens such as Salmonella and Mycobacterium tuberculosis exploit these metabolic networks for immune evasion. By integrating recent advances in host immunometabolism with bacterial pathogenesis, this work not only deciphers critical molecular dialogues at the host-pathogen interface but also identifies novel targetable pathways, offering a conceptual framework for developing innovative therapeutic strategies against persistent and antibiotic-resistant infections.

Keywords:  bacterial infection; glycolytic reprogramming; immunometabolism; macrophage polarization; metabolic signaling

DOI:  https://doi.org/10.3389/fimmu.2026.1783266
Methods Cell Biol. 2026 ;pii: S0091-679X(26)00084-1. [Epub ahead of print]206 1-21

Techniques for monitoring and isolating lipid droplets in cell death.

Fangquan Chen, Rui Kang, Jiao Liu, Daolin Tang.

  Lipid droplets (LDs) are dynamic organelles that not only store energy and regulate metabolism but also serve as key modulators of cell death signaling and disease progression. LDs influence cell fate by buffering lipid peroxidation or releasing fatty acids during ferroptosis, modulating apoptotic protein expression, and facilitating autophagic degradation. Their multifunctional roles are context-dependent and span multiple cell death pathways. In disease, aberrant LD accumulation is closely linked to metabolic disorders, neurodegeneration, cancer therapy resistance, and pathogen infection. Alterations in LD morphology and abundance have emerged as diagnostic indicators. Thus, precise detection and efficient isolation of LDs are critical for elucidating disease mechanisms, advancing targeted therapies, and translating LD biology into clinical applications. This chapter outlines key methodologies-including lipid staining, fluorescent probes, high-content microscopy, density gradient centrifugation, and immunoaffinity purification-for evaluating lipid droplet function and achieving their isolation in the context of cell death.

Keywords:  Cell death; Lipid droplets; Methods

DOI:  https://doi.org/10.1016/bs.mcb.2026.02.016