Resident tissue macrophages are organ-specialized phagocytes responsible for the maintenance and protection of tissue homeostasis. It is well established that tissue diversity is reflected by the heterogeneity of resident tissue macrophage origin and phenotype. However, much less is known about tissue-specific phagocytic and proteolytic macrophage functions. Here, using a quantitative proteomics approach, we identify cathepsins as key determinants of phagosome maturation in primary peritoneum-, lung-, and brain-resident macrophages. The data further uncover cathepsin K (CtsK) as a molecular marker for lung phagosomes required for intracellular protein and collagen degradation. Pharmacological blockade of CtsK activity diminished phagosomal proteolysis and collagenolysis in lung-resident macrophages. Furthermore, profibrotic TGF-β negatively regulated CtsK-mediated phagosomal collagen degradation independently from classical endocytic-proteolytic pathways. In humans, phagosomal CtsK activity was reduced in COPD lung macrophages and non-COPD lung macrophages exposed to cigarette smoke extract. Taken together, this study provides a comprehensive map of how peritoneal, lung, and brain tissue environment shapes phagosomal composition, revealing CtsK as a key molecular determinant of lung phagosomes contributing to phagocytic collagen clearance in lungs.
- MeSH
- fagozomy * metabolismus MeSH
- kathepsin K * metabolismus MeSH
- kolagen metabolismus MeSH
- lidé MeSH
- makrofágy * metabolismus MeSH
- plíce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Dendritic cells (DCs) infected by Francisella tularensis are poorly activated and do not undergo classical maturation process. Although reasons of such unresponsiveness are not fully understood, their impact on the priming of immunity is well appreciated. Previous attempts to explain the behavior of Francisella-infected DCs were hypothesis-driven and focused on events at later stages of infection. Here, we took an alternative unbiased approach by applying methods of global phosphoproteomics to analyze the dynamics of cell signaling in primary DCs during the first hour of infection by Francisella tularensis Presented results show that the early response of DCs to Francisella occurs in phases and that ERK and p38 signaling modules induced at the later stage are differentially regulated by virulent and attenuated ΔdsbA strain. These findings imply that the temporal orchestration of host proinflammatory pathways represents the integral part of Francisella life-cycle inside hijacked DCs.
- MeSH
- buněčné linie MeSH
- dendritické buňky metabolismus mikrobiologie MeSH
- extracelulárním signálem regulované MAP kinasy metabolismus MeSH
- fosforylace MeSH
- Francisella tularensis * MeSH
- mitogenem aktivované proteinkinasy p38 metabolismus MeSH
- myši inbrední C57BL MeSH
- tularemie metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Although dendritic cells (DCs) control the priming of the adaptive immunity response, a comprehensive description of their behavior at the protein level is missing. The introduction of the quantitative proteomic technique of metabolic labeling (SILAC) into the field of DC research would therefore be highly beneficial. To achieve this, we applied SILAC labeling to primary bone marow-derived DCs (BMDCs). These cells combine both biological relevance and experimental feasibility, as their in vitro generation permits the use of (13)C/(15)N-labeled amino acids. Interestingly, BMDCs appear to exhibit a very active arginine metabolism. Using standard cultivation conditions, ∼20% of all protein-incorporated proline was a byproduct of heavy arginine degradation. In addition, the dissipation of (15)N from labeled arginine to the whole proteome was observed. The latter decreased the mass accuracy in MS and affected the natural isotopic distribution of peptides. SILAC-connected metabolic issues were shown to be enhanced by GM-CSF, which is used for the differentiation of DC progenitors. Modifications of the cultivation procedure suppressed the arginine-related effects, yielding cells with a proteome labeling efficiency of ≥90%. Importantly, BMDCs generated according to the new cultivation protocol preserved their resemblance to inflammatory DCs in vivo, as evidenced by their response to LPS treatment.
- MeSH
- arginin metabolismus MeSH
- buňky kostní dřeně metabolismus MeSH
- dendritické buňky metabolismus MeSH
- faktor stimulující granulocyto-makrofágové kolonie metabolismus MeSH
- kultivované buňky MeSH
- molekulární sekvence - údaje MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- prolin metabolismus MeSH
- proteom * MeSH
- sekvence aminokyselin MeSH
- tandemová hmotnostní spektrometrie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Dendritic cells (DCs) serve as the primers of adaptive immunity, which is indispensable for the control of the majority of infections. Interestingly, some pathogenic intracellular bacteria can subvert DC function and gain the advantage of an ineffective host immune reaction. This scenario appears to be the case particularly with so-called stealth pathogens, which are the causative agents of several under-diagnosed chronic diseases. However, there is no consensus how less explored stealth bacteria like Coxiella, Brucella and Francisella cross-talk with DCs. Therefore, the aim of this review was to explore the issue and to summarize the current knowledge regarding the interaction of above mentioned pathogens with DCs as crucial hosts from an infection strategy view. Evidence indicates that infected DCs are not sufficiently activated, do not undergo maturation and do not produce expected proinflammatory cytokines. In some cases, the infected DCs even display immunosuppressive behaviour that may be directly linked to the induction of tolerogenicity favouring pathogen survival and persistence.
- MeSH
- Brucella imunologie fyziologie MeSH
- Coxiella imunologie fyziologie MeSH
- dendritické buňky imunologie mikrobiologie MeSH
- Francisella imunologie fyziologie MeSH
- imunitní únik MeSH
- imunologická tolerance MeSH
- interakce hostitele a patogenu * MeSH
- lidé MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Metalothionein: Jedná se o malý protein s vysokým obsahem cysteinu, jehož sulfhydrylové skupiny mu propůjčují unikátní vlastnosti. V organismu se stará o transport iontů kovů (Zn, Cu), má ale i detoxikační funkci, protože dokáže vázat i toxické těžké kovy (Cd, Pb, Hg aj.). Buňkám pomáhá také zvládat oxidativ-ní stres a reguluje hladinu exprese či enzymatickou aktivitu. Cytostatika na bázi platiny: Tento typ cytostatik patří k nejdéle používaným a nejúčinnějším. Zakládají se na komplexech platiny s obměňováním ligandů. Nejstarší zástupce, cisplatina, má dnes již účinnější analogy (léčiva druhé a třetí generace, karboplatinu a oxaliplatinu, a ve vývoji jsou další), nicméně stále je hojně využívána. U některých nemocných se ale vyskytuje fenomén rezistence nádorových buněk, což samozřejmě snižuje efektivitu léčby. Vznik rezistence a její mechanismy: Rezistence nádorových buněk vůči cytostatikům je komplexní děj, který v sobě zahrnuje několik mechanismů. Z nám známých to jsou především regulace intracelulární koncentrace léku a jeho eflux, komplexace intracelulárními thioly (glutathion, metalothionein) a mechanismy pro reparaci poškozené DNA. Závěr: Výzkumy v této problematice postupují velice rychle a jistě se dočkáme mnoha zlomových poznatků, nejen o zapojení metalothioneinu do vzniku rezistence. Je to jedna z podmínek pro efektivnější zvládnutí léčby a pochopení metabolismu nádorového onemocnění.
Metallothionein: It is a small protein with high content of cysteine, whose sulfhydryl groups give metallothionein its unique properties. In organism metallothionein transports metal ions (Zn, Cu), but it can even bind and eliminate toxic metal ions (Cd, Pb, Hg, etc). Metallothionein helps to cells handle oxidative stress and it regulates level of expression and enzymatic activity. Cytostatics based on platinum: This type of cytostatics belongs to the oldest and most ušed. They are based on platinum complexes with changing ligands. The oldest member of this group, cisplatine, has more effective analogues (platinum complexes of second and third generation, carboplatin and oxaliplatin, and new are under development). Nevertheless cisplatin is still widely ušed. At some patients it developed phenomena of resistance of tumour cells, which of course lower effectiveness of treatment. Induction of resistance and its mechanisms: Resistance of tumour cell to cytostatics is complex process which involves many mechanisms. Some of them are relatively known to us; first of all it is regulation of intracellular concentration of drug and its efflux, complexation by intracellular thiols (glutathione and metallothionein) and mechanisms for repairing of damaged DNA. Conclusion: Research of this problems progresses very quickly and new information wait ahead us not only about influence of metallothionein on induction of resistance. It is one of preconditions for more effective treatment and understanding of metabolism of tumour disease.
