Pemphigus Vulgaris (PV) is a life-threatening autoimmune disease manifested with blisters in the skin and mucosa and caused by autoantibodies against adhesion protein desmoglein-3 (Dsg3) expressed in epithelial membrane linings of these tissues. Despite many studies, the pathogenesis of PV remains incompletely understood. Recently we have shown Dsg3 plays a role in regulating the yes-associated protein (YAP), a co-transcription factor and mechanical sensor, and constraining reactive oxygen species (ROS). This study investigated the effect of PV sera as well as the anti-Dsg3 antibody AK23 on these molecules. We detected elevated YAP steady-state protein levels in PV cells surrounding blisters and perilesional regions and in keratinocytes treated with PV sera and AK23 with concomitant transient ROS overproduction. Cells treated with hydrogen peroxide also exhibited augmented nuclear YAP accompanied by reduction of Dsg3 and α-catenin, a negative regulator of YAP. As expected, transfection of α-catenin-GFP plasmid rendered YAP export from the nucleus evoked by hydrogen peroxide. In addition, suppression of total YAP was observed in hydrogen peroxide treated cells exposed to antioxidants with enhanced cell-cell adhesion being confirmed by decreased fragmentation in the dispase assay compared to hydrogen peroxide treatment alone. On the other hand, the expression of exogenous YAP disrupted intercellular junction assembly. In contrast, YAP depletion resulted in an inverse effect with augmented expression of junction assembly proteins, including Dsg3 and α-catenin capable of abolishing the effect of AK23 on Dsg3 expression. Finally, inhibition of other kinase pathways, including p38MAPK, also demonstrated suppression of YAP induced by hydrogen peroxide. Furthermore, antioxidant treatment of keratinocytes suppressed PV sera-induced total YAP accumulation. In conclusion, this study suggests that oxidative stress coupled with YAP dysregulation attributes to PV blistering, implying antioxidants may be beneficial in the treatment of PV.
- MeSH
- adaptorové proteiny signální transdukční genetika metabolismus MeSH
- alfa-katenin metabolismus MeSH
- antioxidancia farmakologie terapeutické užití MeSH
- autoprotilátky krev imunologie metabolismus MeSH
- buněčná adheze účinky léků imunologie MeSH
- buněčné linie MeSH
- desmoglein 3 imunologie metabolismus MeSH
- genový knockdown MeSH
- keratinocyty MeSH
- lidé MeSH
- MAP kinasový signální systém účinky léků imunologie MeSH
- oxidační stres účinky léků imunologie MeSH
- pemfigus krev farmakoterapie imunologie patologie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- studie případů a kontrol MeSH
- transkripční faktory genetika metabolismus MeSH
- ústní sliznice imunologie patologie MeSH
- zdraví dobrovolníci pro lékařské studie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
- práce podpořená grantem MeSH
Amine-coated biodegradable materials based on synthetic polymers have a great potential for tissue remodeling and regeneration because of their excellent processability and bioactivity. In the present study, we have investigated the influence of various chemical compositions of amine plasma polymer (PP) coatings and the influence of the substrate morphology, represented by polystyrene culture dishes and polycaprolactone nanofibers (PCL NFs), on the behavior of vascular smooth muscle cells (VSMCs). Although all amine-PP coatings improved the initial adhesion of VSMCs, 7-day long cultivation revealed a clear preference for the coating containing about 15 at.% of nitrogen (CPA-33). The CPA-33 coating demonstrated the ideal combination of good water stability, a sufficient amine group content, and favorable surface wettability and morphology. The nanostructured morphology of amine-PP-coated PCL NFs successfully slowed the proliferation rate of VSMCs, which is essential in preventing restenosis of vascular replacements in vivo. At the same time, CPA-33-coated PCL NFs supported the continuous proliferation of VSMCs during 7-day long cultivation, with no significant increase in cytokine secretion by RAW 264.7 macrophages. The CPA-33 coating deposited on biodegradable PCL NFs therefore seems to be a promising material for manufacturing small-diameter vascular grafts, which are still lacking on the current market.
- MeSH
- aminy škodlivé účinky chemie imunologie farmakologie MeSH
- biokompatibilní potahované materiály škodlivé účinky chemie farmakologie MeSH
- buněčná adheze účinky léků imunologie MeSH
- fotoelektronová spektroskopie MeSH
- krevní plazma chemie imunologie MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- makrofágy účinky léků metabolismus MeSH
- myocyty hladké svaloviny účinky léků metabolismus MeSH
- myši MeSH
- nanovlákna škodlivé účinky chemie MeSH
- polyestery chemie MeSH
- polymery škodlivé účinky chemie farmakologie MeSH
- povrchové vlastnosti účinky léků MeSH
- proliferace buněk účinky léků MeSH
- RAW 264.7 buňky MeSH
- svaly hladké cévní cytologie účinky léků růst a vývoj MeSH
- tkáňové podpůrné struktury škodlivé účinky chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Extensive exploitation of titanium dioxide nanoparticles (TiO2NPs) augments rapid release into the marine environment. When in contact with the body fluids of marine invertebrates, TiO2NPs undergo a transformation and adhere various organic molecules that shape a complex protein corona prior to contacting cells and tissues. To elucidate the potential extracellular signals that may be involved in the particle recognition by immune cells of the sea urchin Paracentrotus lividus, we investigated the behavior of TiO2NPs in contact with extracellular proteins in vitro. Our findings indicate that TiO2NPs are able to interact with sea urchin proteins in both cell-free and cell-conditioned media. The two-dimensional proteome analysis of the protein corona bound to TiO2NP revealed that negatively charged proteins bound preferentially to the particles. The main constituents shaping the sea urchin cell-conditioned TiO2NP protein corona were proteins involved in cellular adhesion (Pl-toposome, Pl-galectin-8, Pl-nectin) and cytoskeletal organization (actin and tubulin). Immune cells (phagocytes) aggregated TiO2NPs on the outer cell surface and within well-organized vesicles without eliciting harmful effects on the biological activities of the cells. Cells showed an active metabolism, no oxidative stress or caspase activation. These results provide a new level of understanding of the extracellular proteins involved in the immune-TiO2NP recognition and interaction in vitro, confirming that primary immune cell cultures from P. lividus can be an optional model for swift and efficient immune-toxicological investigations.
- MeSH
- buněčná adheze imunologie MeSH
- fagocyty imunologie MeSH
- galektiny imunologie MeSH
- glykoproteiny imunologie MeSH
- ježovky imunologie MeSH
- nanočástice aplikace a dávkování MeSH
- nektiny imunologie MeSH
- Paracentrotus imunologie MeSH
- proteinová korona imunologie MeSH
- proteom imunologie MeSH
- titan imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- ingesce, nitrobuněčné zabíjení, porucha fagocytózy,
- MeSH
- buněčná adheze fyziologie imunologie MeSH
- chemotaxe fyziologie imunologie MeSH
- fagocytóza * fyziologie imunologie MeSH
- lidé MeSH
- NADPH-oxidasy fyziologie imunologie MeSH
- neutrofily fyziologie imunologie MeSH
- opsonizace fyziologie imunologie MeSH
- přirozená imunita fyziologie MeSH
- transendoteliální a transepiteliální migrace fyziologie imunologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- buněčná adheze genetika imunologie MeSH
- endoplazmatické retikulum genetika imunologie metabolismus MeSH
- finanční podpora výzkumu jako téma MeSH
- fosforylace MeSH
- kadheriny genetika imunologie metabolismus MeSH
- kalmodulin genetika imunologie MeSH
- kalretikulin genetika MeSH
- lidé MeSH
- signální transdukce genetika imunologie MeSH
- vinkulin genetika imunologie metabolismus MeSH
- Check Tag
- lidé MeSH
