Browsing by Subject "Macrophage"
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- PublicationOpen AccessA genetically encoded IL-1β BRET sensor to monitor inflammasome activity(Asociación Americana de Inmunología, 2012) Compan, Vincent; Baroja-Mazo, Alberto; Bragg, Laricia; Verkhratsky, Alexei; Perroy, Julie; Pelegrin, Pablo; Bioquímica y Biología Molecular B e InmunologíaInflammation is fundamental for protecting the organism against infection and injury. However, a failure to control immune response results in chronic inflammation and several associated disorders such as pain and loss of function. Initiation of inflammation is orchestrated by cytokines, among which interleukin (IL)-1β is particularly important. IL- 1β is synthesized as an inactive protein that has to be processed by the inflammasome to generate the mature bioactive form. Conventional techniques cannot monitor IL-1β activation with high spatial and temporal resolution. Here, we present a ratiometric biosensor that allows monitoring IL-1β processing in real-time, with a temporal resolution of seconds and with a single cell spatial resolution. Using this sensor, we describe for the first time the kinetic of the inflammasome activity in living macrophages. With this new probe we also demonstrated that the pro-IL-1β processing occurs all over the cytoplasm.
- PublicationOpen AccessAutophagy and the regulation of the immune response(2012-12) Macián, Fernando; Valdor Alonso, Rut; Bioquímica y Biología Molecular B e InmunologíaAutophagy is a highly conserved mechanism of lysosomal-mediated protein degradation that plays a crucial role in maintaining cellular homeostasis by recycling amino acids, reducing the amount of damaged proteins and regulating protein levels in response to extracellular signals. In the last few years specific functions for different forms of autophagy have been identified in many tissues and organs. In the Immune System, autophagy functions range from the elimination infectious agents and the modulation of the inflammatory response, to the selection of antigens for presentation and the regulation of T cell homeostasis and activation. Here, we review the recent advances that have allowed us to better understand why autophagy is a crucial process in the regulation of the innate and adaptive immune responses.
- PublicationOpen AccessGasdermin D mediates a fast transient release of ATP after NLRP3 inflammasome activation before ninjurin 1-induced lytic cell death(Cell Press, 2025) Schachter, Julieta; Guijarro, Adriana; Angosto-Bazarra, Diego; Pinilla, Miriam; Hurtado-Navarro, Laura; Meunier, Etienne; Perez-Oliva, Ana Belen; Schwarzbaum, Pablo J; Pelegrín Vivancos, Pablo; Pinilla Marquinez, Míriam; Bioquímica y Biología Molecular B e InmunologíaPyroptosis is a lytic cell death triggered by the cleavage of gasdermin (GSDM) proteins and subsequent pore formation by the N-terminal domain oligomerization in the plasma membrane. GSDMD is cleaved by caspase-1/-4/-5/-11 upon inflammasome activation and mediates interleukin (IL)-1β and IL-18 release. GSDMD pores favor ninjurin 1 (NINJ1)-induced plasma membrane rupture and cell death. Here, we demonstrate that GSDMD mediates early ATP release upon NLRP3 inflammasome activation independently of NINJ1, occurring before IL-1β release and cell death and constituting an early danger signal. The release of ATP is a transient signal terminated before the cells continue to permeabilize and die. The different N termini of GSDMA to -E are also able to release ATP and induce monocyte migration toward pyroptotic cells. This study reveals ATP release as an early and transient danger signal depending on GSDMD plasma membrane permeabilization, independently of the late stages of lytic cell death.
- PublicationRestrictedMembrane vesicles for nanoencapsulated sulforaphane increased their anti-inflammatory role on an In vitro human macrophage model.(MDPI, 2022-02-09) Yepes-Molina, Lucía; Pérez-Jiménez, María Isabel; Martínez-Esparza Alvargonzález, María Concepción; Teruel Puche, José Antonio; Ruiz Alcaraz, Antonio José; García Peñarrubia, María del Pilar; Carvajal, Micaela; Bioquímica y Biología Molecular B e InmunologíaAt present, there is a growing interest in finding new non‐toxic anti‐inflammatory drugs to treat inflammation, which is a key pathology in the development of several diseases with considerable mortality. Sulforaphane (SFN), a bioactive compound derived from Brassica plants, was shown to be promising due to its anti‐inflammatory properties and great potential, though its actual clinical use is limited due to its poor stability and bioavailability. In this sense, the use of nanocarriers could solve stability‐related problems. In the current study, sulforaphane loaded into membrane vesicles derived from broccoli plants was studied to determine the anti‐inflammatory potential in a human‐macrophage‐like in vitro cell model under both normal and inflammatory conditions. On the one hand, the release of SFN from membrane vesicles was modeled in vitro, and two release phases were stabilized, one faster and the other slower due to the interaction between SFN and membrane proteins, such as aquaporins. Furthermore, the anti‐inflammatory action of sulforaphane‐loaded membrane vesicles was demonstrated, as a decrease in interleukins crucial for the development of nflammation, such as TNF‐α, IL‐1β and IL‐6, was observed. Furthermore, these results also showed that membrane vesicles by themselves had anti‐inflammatory properties, opening the possibility of new lines of research to study these vesicles, not only as carriers but also as active compounds.
PublicationRestrictedMicafungin enhances the human macrophage response to Candida albicans through β-glucan exposure(American Society for Microbiology, 2018-04-26) Guirao-Abad, José Pedro; Sánchez-Fresneda, Ruth; Machado, Francisco; Argüelles, Juan Carlos; Martínez-Esparza Alvargonzález, María Concepción; Bioquímica y Biología Molecular B e InmunologíaMicafungin belongs to the antifungal family of echinocandins, which act as noncompetitive inhibitors of the fungal cell wall β-1,3-d-glucan synthase. Since Candida albicans is the most prevalent pathogenic fungus in humans, we study the involvement of micafungin in the modulation of the inflammatory response developed by human tissue macrophages against C. albicans The MIC for micafungin was 0.016 μg/ml on the C. albicans SC5314 standard strain. Micafungin induced a drastic reduction in the number of exponential SC5314 viable cells, with the fungicidal effect being dependent on the cellular metabolic activity. Notably, micafungin also caused a structural remodelling of the cell wall, leading to exposure of the β-glucan and chitin content on the external surface. At the higher doses used (0.05 μg/ml), the antifungal also induced the blowing up of budding yeasts. In addition, preincubation with micafungin before exposure to human tissue macrophages enhanced the secretion of tumor necrosis factor alpha (TNF-α), interleukin-17A (IL-17A), and IL-10 cytokines. Our results strongly suggest that in C. albicans treatment with micafungin, in addition to having the expected toxic antifungal effect, it potentiates the immune response, improving the interaction and activation of human macrophages, probably through the unmasking of β-glucans on the cell wall surface.- PublicationOpen AccessMitochondrial respiratory-chain adaptations in macrophages contribute to antibacterial host defense(Nature, 2016) Garaude, Johan; Acín-Pérez, Rebeca; Martínez-Cano, Sarai; Enamorado, Michael; Ugolini, Matteo; Nistal-Villan, Estanislao; Hervás-Stubbs, Sandra; Pelegrin, Pablo; Sander, Leif E.; Enríquez, José A.; Sancho, David; Bioquímica y Biología Molecular B e InmunologíaThe mitochondrial electron transport chain (ETC) is a metabolic hub whose adaptations accompany fuel source fluctuations, stress responses, and innate immune signals to ensure optimal cellular functions. Macrophages tightly scale their core metabolism upon activation by innate immune receptors but the precise regulation of the ETC upon pathogen recognition and its functional implications are currently unknown. Here we show that innate immune sensing of live bacteria by macrophages elicits transient ETC adaptations that is characterized by a decrease assembly of complex I (CI) and CI-containing supercomplexes and by a switch in the relative contribution of complexes I and II to mitochondrial respiration. This is mediated by the phagosomal nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase and the reactive oxygen species (ROS)-dependent Src-family tyrosine-kinase Fgr and required Toll-like receptor (TLR) signalling and the NOD-like receptor (NLR) family, pyrin domain–containing protein 3 (NLRP3) inflammasome, both connected to bacterial viability-specific immune responses. Consistently, the inhibition of CII in E. coli infected mice decreases IL-1b and increases IL-10 serum-levels to those found in mice treated with dead bacteria and impairs control of bacteria. We thus identify the innate immune receptor-mediated ETC adaptations as an early immune-metabolic checkpoint that potentially adjusts innate immune responses during bacterial infection.
- PublicationOpen AccessNLRP3 lacking the leucine-rich repeat domain can be fully activated via the canonical inflammasome pathway(Nature, 2018) Hafner-Bratkovic, Iva; Susjan, Petra; Lainscek, Dusko; Cerovic, Kosta; Kadunc, Lucija; Angosto-Bazarra, Diego; Pelegrin, Pablo; Jerala, Roman; Tapia Abellán, Ana; Bioquímica y Biología Molecular B e InmunologíaNLRP3 is a cytosolic sensor triggered by different pathogen- and self-derived signals that plays a central role in a variety of pathological conditions, including sterile inflammation. The leucine- rich repeat domain is present in several innate immune receptors, where it is frequently responsible for sensing danger signals and regulation of activation. We show by reconstitution of truncated and chimeric variants into NLRP3-/- macrophages that the leucine-rich repeat domain is dispensable for activation and self-regulation of NLRP3 by several different triggers. The pyrin domain on the other hand is required to maintain NLRP3 in the inactive conformation. A fully responsive minimal NLRP3 truncation variant reconstituted peritonitis in NLRP3-/- mice. We demonstrate that in contrast to pathogen-activated NLRC4, the constitutively active NLRP3 molecule cannot engage wild-type NLRP3 molecules in a self-catalytic oligomerization. This lack of signal amplification is likely a protective mechanism to decrease sensitivity to endogenous triggers to impede autoinflammation.
- PublicationOpen AccessPotential of sulforaphane and broccoli membrane vesicles as regulators of M1/M2 human macrophage activity(2022-09-22) García-Peñaranda, Andrea; García-Ibáñez, Paula; Yepes-Molina, Lucía; Carvajal, Micaela; Ruiz Alcaraz, Antonio José; Moreno, Diego A.; García Peñarrubia, María del Pilar; Martínez-Esparza Alvargonzález, María Concepción; Ramírez Pávez, Tamara Nadira; Bioquímica y Biología Molecular B e Inmunología; Facultad de Biología
- PublicationOpen AccessPriming is dispensable for NLRP3 inflammasome activation in human monocytes in vitro(Frontiers Media, 2020-09-30) Martín Sánchez, María Rosario Fátima; Gritsenko, Anna; Yu, Shi; Díaz del Olmo, Inés; Nichols, Eva María; Davis, Daniel M.; Brough, David; López Castejón, Gloria; FarmacologíaInterleukin (IL)-18 and IL-1b are potent pro-inflammatory cytokines that contribute toinflammatory conditions such as rheumatoid arthritis and Alzheimer’s disease. They areproduced as inactive precursors that are activated by large macromolecular complexescalled inflammasomes upon sensing damage or pathogenic signals. NLRP3inflammasome activation is regarded to require a priming step that causes NLRP3 andIL-1b gene upregulation, and also NLRP3 post-translational licencing. A subsequentactivation step leads to the assembly of the complex and the cleavage of pro-IL-18 andpro-IL-1b by caspase-1 into their mature forms, allowing their release. Here we show thathuman monocytes, but not monocyte derived macrophages, are able to form canonicalNLRP3 inflammasomes in the absence of priming. NLRP3 activator nigericin caused theprocessing and release of constitutively expressed IL-18 in an unprimed setting. This wasmediated by the canonical NLRP3 inflammasome that was dependent on K + and Cl−efflux and led to ASC oligomerization, caspase-1 and Gasdermin-D (GSDMD) cleavage.IL-18 release was impaired by the NLRP3 inhibitor MCC950 and by the absence ofNLRP3, but also by deficiency of GSDMD, suggesting that pyroptosis is the mechanism ofrelease. This work highlights the readiness of the NLRP3 inflammasome to assemble inthe absence of priming in human monocytes and hence contribute to the very early stagesof the inflammatory response when IL-1b has not yet been produced. It is important toconsider the unprimed setting when researching the mechanisms of NLRP3 activation, asto not overshadow the pathways that occur in the absence of priming stimuli, which mightonly enhance this response.
- ItemOpen AccessRecent progress in macrophage- mediated tendon injury and healing(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2026) Yining Zhang; Yanzhao Dong; Xiaodi Zou; Ahmad Alhaskawi; Fangyu Yi; Sohaib Hasan Abdullah Ezzi; Vishnu Goutham Kota; Mohamed Hasan Abdulla Hasan Abdulla; Haiying Zhou; Alenikova Olga; Sahar Ahmed Abdalbary; Jinhui Liang; Hui Lu; Weijie Zhou; Biología Celular e HistologíaRecently, the role of macrophages in tendon repair has received increased attention. These cells are versatile, playing multiple roles, such as defending the host, engulfing debris, producing growth factors, and releasing substances that can both promote and alleviate inflammation. Within the scope of tendon repair and tendinopathy resolution, macrophages are essential contributors. However, the current understanding of macrophage involvement in tendon healing remains limited. Hence, understanding macrophages' impact on tendon healing is of considerable importance for devising innovative treatment approaches. It is crucial to examine the precise role that macrophages play in tendon recovery, as it provides new understanding that can propel both research and the development of therapeutic methods aimed at enhancing tendon recovery moving forward
- PublicationOpen AccessThe significance of M1 macrophage should be highlighted in peripheral nerve regeneration(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2023) Feng, Ruiqin; Zhang, PeixunMacrophage influences peripheral nerve regeneration. According to the classical M1/M2 paradigm, the M1 macrophage is an inhibitor of regeneration, while the M2 macrophage is a promoter. However, several studies have shown that M1 macrophages are indispensable for peripheral nerve repair and facilitate many critical processes in axonal regeneration. In this review, we summarized the information on macrophage polarization and focused on the activities of M1 macrophages in regeneration. We also provided some examples where the macrophage phenotypes were regulated to help regeneration. We argued that the coordination of both macrophage phenotypes might be effective in peripheral nerve repair, and a more comprehensive view of macrophages might contribute to macrophage-based immunomodulatory therapies.
- PublicationOpen AccessThe total electric charge and time of application of galvanic currents to macrophages can optimize the release of IL-1β with low cell death(Nature Research, 2024-12-28) Peñín Franc, Alejandro; García Vidal, José Antonio; Gómez, Ana Isabel; Escolar Reina, Pilar; Medina Mirapeix, Francesc; Pelegrín Vivancos, Pablo; FisioterapiaGalvanic current has been emerging as a novel therapy to regenerate chronic tissue lesions, including musculoskeletal and dermatological lesions. Recently, the NLRP3 inflammasome and IL-1β release have been identified as a signaling pathway triggered upon galvanic current application. However, the parameters for the clinical application of galvanic current remain subjective to the experience of the facultative in charge. In this study we used an in vitro model of macrophage culture and application of different combinations of the parameters of galvanic current to study IL-1β production and cell death. Increasing electric charge of galvanic current induces the release of IL-1β, but electric charges equal or higher to 144 mC also increase cell death. The release of IL-1β have a substantial variation within different electric charge of galvanic currents, being increased by decreasing the current and increasing the time of current application. Within the range of current intensities studied, the most optimal protocol for maximizing IL-1β release without inducing cell death was identified at electric charges equal to or near 144 mC, applied over a total duration of approximately 25 s. Our findings lay the groundwork for future in vivo studies assessing different electric charge of galvanic current, with the aim of yielding clinically relevant outcomes.
- PublicationOpen AccessTNF induces pathogenic programmed macrophage necrosis in Tuberculosis through a mitochondrial- lysosomal-endoplasmic reticulum circuit(Cell Press, 2019-08-29) Roca Soler, Francisco José; Whitworth, Laura J.; Redmond, Sarah; Jones, Ana A.; Ramakrishnan, Lalita; Bioquímica y Biología Molecular B e InmunologíaNecrosis of infected macrophages constitutes a critical pathogenetic event in tuberculosis by releasing mycobacteria into the growth-permissive extracellular environment. In zebrafish infected with Mycobacterium marinum or Mycobacterium tuberculosis, excess tumor necrosis factor triggers programmed necrosis of infected macrophages through the production of mitochondrial reactive oxygen species (ROS) and the participation of cyclophilin D, a component of the mitochondrial permeability transition pore. Here, we show that this necrosis pathway is not mitochondrion-intrinsic but results from an inter-organellar circuit initiating and culminating in the mitochondrion. Mitochondrial ROS induce production of lysosomal ceramide that ultimately activates the cytosolic protein BAX. BAX promotes calcium flow from the endoplasmic reticulum into the mitochondrion through ryanodine receptors, and the resultant mitochondrial calcium overload triggers cyclophilin-D-mediated necrosis. We identify ryanodine receptors and plasma membrane L-type calcium channels as druggable targets to intercept mitochondrial calcium overload and necrosis of mycobacterium-infected zebrafish and human macrophages.