Tetherin/BST-2 is an antiviral protein that blocks the release of enveloped viral particles by linking them to the membrane of producing cells. At first, BST-2 genes were described only in humans and other mammals. Recent work identified BST-2 orthologs in nonmammalian vertebrates, including birds. Here, we identify the BST-2 sequence in domestic chicken (Gallus gallus) for the first time and demonstrate its activity against avian sarcoma and leukosis virus (ASLV). We generated a BST-2 knockout in chicken cells and showed that BST-2 is a major determinant of an interferon-induced block of ASLV release. Ectopic expression of chicken BST-2 blocks the release of ASLV in chicken cells and of human immunodeficiency virus type 1 (HIV-1) in human cells. Using metabolic labeling and pulse-chase analysis of HIV-1 Gag proteins, we verified that chicken BST-2 blocks the virus at the release stage. Furthermore, we describe BST-2 orthologs in multiple avian species from 12 avian orders. Previously, some of these species were reported to lack BST-2, highlighting the difficulty of identifying sequences of this extremely variable gene. We analyzed BST-2 genes in the avian orders Galliformes and Passeriformes and showed that they evolve under positive selection. This indicates that avian BST-2 is involved in host-virus evolutionary arms races and suggests that BST-2 antagonists exist in some avian viruses. In summary, we show that chicken BST-2 has the potential to act as a restriction factor against ASLV. Characterizing the interaction of avian BST-2 with avian viruses is important in understanding innate antiviral defenses in birds.IMPORTANCE Birds are important hosts of viruses that have the potential to cause zoonotic infections in humans. However, only a few antiviral genes (called viral restriction factors) have been described in birds, mostly because birds lack counterparts of highly studied mammalian restriction factors. Tetherin/BST-2 is a restriction factor, originally described in humans, that blocks the release of newly formed virus particles from infected cells. Recent work identified BST-2 in nonmammalian vertebrate species, including birds. Here, we report the BST-2 sequence in domestic chicken and describe its antiviral activity against a prototypical avian retrovirus, avian sarcoma and leukosis virus (ASLV). We also identify BST-2 genes in multiple avian species and show that they evolve rapidly in birds, which is an important indication of their relevance for antiviral defense. Analysis of avian BST-2 genes will shed light on defense mechanisms against avian viral pathogens.
- MeSH
- antigen stromálních buněk kostní dřeně genetika imunologie MeSH
- buněčné linie MeSH
- fibroblasty imunologie virologie MeSH
- Galliformes genetika imunologie virologie MeSH
- genové produkty gag - virus lidské imunodeficience genetika imunologie MeSH
- HEK293 buňky MeSH
- HIV-1 genetika imunologie MeSH
- interakce hostitele a patogenu genetika imunologie MeSH
- lidé MeSH
- molekulární evoluce * MeSH
- Passeriformes genetika imunologie virologie MeSH
- ptačí proteiny genetika imunologie MeSH
- ptačí sarkom genetika imunologie virologie MeSH
- regulace genové exprese MeSH
- replikace viru MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- sekvenční seřazení MeSH
- selekce (genetika) MeSH
- signální transdukce MeSH
- uvolnění viru z buňky MeSH
- viry ptačího sarkomu genetika imunologie patogenita MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, N.I.H., Intramural MeSH
Viral and cellular double-stranded RNA (dsRNA) is recognized by cytosolic innate immune sensors, including RIG-I-like receptors. Some cytoplasmic dsRNA is commonly present in cells, and one source is mitochondrial dsRNA, which results from bidirectional transcription of mitochondrial DNA (mtDNA). Here we demonstrate that Trp53 mutant mouse embryonic fibroblasts contain immune-stimulating endogenous dsRNA of mitochondrial origin. We show that the immune response induced by this dsRNA is mediated via RIG-I-like receptors and leads to the expression of type I interferon and proinflammatory cytokine genes. The mitochondrial dsRNA is cleaved by RNase L, which cleaves all cellular RNA including mitochondrial mRNAs, increasing activation of RIG-I-like receptors. When mitochondrial transcription is interrupted there is a subsequent decrease in this immune-stimulatory dsRNA. Our results reveal that the role of p53 in innate immunity is even more versatile and complex than previously anticipated. Our study, therefore, sheds new light on the role of endogenous RNA in diseases featuring aberrant immune responses.
- MeSH
- adenosindeaminasa nedostatek genetika imunologie MeSH
- DEAD box protein 58 genetika imunologie MeSH
- dvouvláknová RNA genetika imunologie MeSH
- embryo savčí MeSH
- endoribonukleasy genetika imunologie MeSH
- fibroblasty cytologie imunologie MeSH
- genetická transkripce MeSH
- IFIH1 genetika imunologie MeSH
- interferonový regulační faktor 7 genetika imunologie MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- nádorový supresorový protein p53 nedostatek genetika imunologie MeSH
- přirozená imunita genetika MeSH
- proteiny genetika imunologie MeSH
- RNA mitochondriální genetika imunologie MeSH
- transfekce MeSH
- transportní proteiny genetika imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
OBJECTIVES: IL-35 is a member of the IL-12 family consisting of p35/IL-12a and EBI3/IL-27b subunits. IL-35 exerts immunomodulatory activities in experimental and human autoimmune inflammatory conditions. Our aim was to assess IL-35 expression in the skin and circulation of SSc patients and to characterize its potential association with SSc-related features. METHODS: Expression of IL-35 in skin and dermal fibroblasts was quantified by quantitative PCR, immunohistochemistry and immunofluorescence. Serum levels of IL-35 (by ELISA), CRP (by turbidimetry), ANA (by immunofluorescence) and autoantibodies of the ENA complex (by immunoblot) were measured in 40 SSc patients. Serum IL-35 was determined in 40 age- and sex-matched healthy controls. RESULTS: IL-35 expression was increased in SSc skin and dermal fibroblasts in a TGF-β-dependent manner. IL-35 induced an activated phenotype in resting fibroblasts and enhanced the release of collagen. IL-35 serum levels were increased in patients with SSc compared with healthy controls [median 83.9 (interquartile range 45.1-146.1) vs 36.2 (interquartile range 17.2-49.4) pg/ml, P < 0.0001]. Serum IL-35 was negatively correlated with disease duration (r = -0.4339, P = 0.0052). In line with this finding, serum IL-35 was increased in patients with an early SSc pattern on capillaroscopy assessment compared with those with active and late SSc patterns. CONCLUSION: The present study demonstrates overexpression of IL-35 in SSc skin, dermal fibroblasts and serum. TGF-β induces IL-35, which in turn activates resting fibroblasts and enhances the release of collagen, thereby contributing to aberrant TGF-β signalling in SSc. Increased serum IL-35 is associated with early, inflammatory stages of SSc.
- MeSH
- dospělí MeSH
- fibroblasty imunologie MeSH
- interleukiny biosyntéza krev genetika MeSH
- kolagen biosyntéza MeSH
- kultivované buňky MeSH
- kůže imunologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA genetika MeSH
- senioři MeSH
- studie případů a kontrol MeSH
- systémová sklerodermie imunologie MeSH
- transformující růstový faktor beta imunologie MeSH
- upregulace imunologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- biopsie metody normy využití MeSH
- buňky stromatu imunologie patologie MeSH
- cévy imunologie metabolismus růst a vývoj MeSH
- cílená molekulární terapie * metody využití MeSH
- fibroblasty cytologie imunologie účinky léků MeSH
- geny nádorové genetika MeSH
- ischemie metabolismus MeSH
- karcinom MeSH
- lidé MeSH
- metastázy nádorů MeSH
- mnohočetná léková rezistence imunologie účinky léků MeSH
- mutace MeSH
- nádorové biomarkery MeSH
- nádory * diagnóza etiologie imunologie MeSH
- progrese nemoci MeSH
- proliferace buněk MeSH
- recidiva MeSH
- signální transdukce účinky léků MeSH
- tumor infiltrující lymfocyty imunologie účinky léků MeSH
- výzkum MeSH
- Check Tag
- lidé MeSH
Complex regulation of the wound healing process involves multiple interactions among stromal tissue cells, inflammatory cells, and the extracellular matrix. Low molecular weight hyaluronan (LMW HA) derived from the degradation of high molecular weight hyaluronan (HMW HA) is suggested to activate cells involved in wound healing through interaction with HA receptors. In particular, receptor CD44 is suggested to mediate cell response to HA of different MW, being the main cell surface HA receptor in stromal tissue and immune cells. However, the response of dermal fibroblasts, the key players in granulation tissue formation within the wound healing process, to LMW HA and their importance for the activation of immune cells is unclear. In this study we show that LMW HA (4.3kDa) induced pro-inflammatory cytokine IL-6 and chemokines IL-8, CXCL1, CXCL2, CXCL6 and CCL8 gene expression in normal human dermal fibroblasts (NHDF) that was further confirmed by increased levels of IL-6 and IL-8 in cell culture supernatants. Conversely, NHDF treated by HMW HA revealed a tendency to decrease the gene expression of these cytokine and chemokines when compared to untreated control. The blockage of CD44 expression by siRNA resulted in the attenuation of IL-6 and chemokines expression in LMW HA treated NHDF suggesting the involvement of CD44 in LMW HA mediated NHDF activation. The importance of pro-inflammatory mediators produced by LMW HA triggered NHDF was evaluated by significant activation of blood leukocytes exhibited as increased production of IL-6 and TNF-α. Conclusively, we demonstrated a pro-inflammatory response of dermal fibroblasts to LMW HA that was transferred to leukocytes indicating the significance of LMW HA in the inflammatory process development during the wound healing process.
- MeSH
- antigeny CD44 metabolismus MeSH
- chemokiny biosyntéza genetika MeSH
- fibroblasty účinky léků imunologie MeSH
- interleukin-6 biosyntéza genetika metabolismus MeSH
- interleukin-8 biosyntéza MeSH
- kyselina hyaluronová farmakologie MeSH
- leukocyty účinky léků metabolismus MeSH
- lidé MeSH
- malá interferující RNA metabolismus MeSH
- mediátory zánětu metabolismus MeSH
- messenger RNA genetika metabolismus MeSH
- molekulová hmotnost MeSH
- přirozená imunita účinky léků genetika MeSH
- signální transdukce účinky léků genetika MeSH
- škára cytologie MeSH
- TNF-alfa biosyntéza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Immunocompatibility of gelatin-based hydrogels to be applied as implant coatings for local regenerative treatment has been studied. First, the bio- and immuno-acceptability of the methacrylamide-modified gelatin hydrogels per se was screened. The results indicated that the hydrogels support cell growth. Metabolic activity of normal cells and permanent cell lines representing various cell types (endothelial, epithelial, fibroblast, and monocyte/macrophage) cultivated on the gelatin hydrogels was moderately lower compared to cells cultivated on tissue culture plastic. The cells cultivated on the hydrogels produced identical cytokines as the control cells although at lower levels. Importantly, no inflammatory activity, measured by nitric oxide and pro-inflammatory cytokine (IL-1α, IL-6, and TNFα) production, was observed in peritoneal cells and monocyte/macrophage RAW 264.7 cell line cultivated on the hydrogels. Finally, polyimide (PI) implantable membranes were surface-modified with gelatin hydrogels and screened for their in vivo immunocompatibility. Their histological examination performed after subcutaneous implantation in mice produced a sound proof of immunoacceptability. Normal tissue repair, mild cellular infiltration and edema mainly induced by the surgery were observed after 2 and 6 days. No adverse tissue responses were induced by the implants. Analysis performed after 4 and 9 weeks indicated areas of foreign body granuloma without formation of a fibrous capsule.
- MeSH
- akrylamidy chemie imunologie MeSH
- biokompatibilní materiály chemie metabolismus MeSH
- buněčné linie MeSH
- cytokiny imunologie MeSH
- fibroblasty cytologie imunologie MeSH
- hydrogely chemie metabolismus MeSH
- lidé MeSH
- makrofágy cytologie imunologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- proliferace buněk MeSH
- protézy a implantáty MeSH
- regenerativní lékařství MeSH
- želatina chemie imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Úvod: Macleaya cordata (Willd.) obsahuje kvarterní benzo[c]fenanthridinové alkaloidy sanguinarin a chelerythrin. Alkaloidový extrakt M. cordata (MCE) je znám pro své antimikrobiální a protizánětlivé účinky a je užíván jako aktivní složka v přípravcích ústní hygieny. Cíl práce: Práce se zabývá ověřením účinku MCE na zánětlivou reakci a oxidační stres vyvolaný v lidských gingiválních fibroblastech působením bakteriálního lipopolysacharidu (LPS). Materiál a metodika: Účinek MCE (0,01–10 μg/ml) na viabilitu lidských gingiválních fibroblastů byl hodnocen stanovením inkorporace neutrální červeně do buněk. Netoxické koncentrace extraktu (0,25 a 0,5 μg/ml) byly použity pro studium vlivu MCE na hladinu redukovaného gluthathionu (GSH), produkci reaktivních kyslíkových sloučenin (ROS), expresi cyklooxygenázy-2 (COX-2) a interleukinu-6 (IL-6) v buňkách vystavených působení LPS. Výsledky: Působením obsahových složek MCE došlo u gingiválních fibroblastů vystaveným LPS ke zvýšení intracelulárního GSH, snížení produkce ROS a k redukci exprese IL-6 a COX-2. Závěr: Alkaloidový extrakt M. cordata potlačil zánětlivou reakci a oxidační stres v lidských gingiválních fibroblastech a může být prakticky využit při ošetření parodontitid a alveolitid.
Introduction: Macleaya cordata (Willd.) contains quaternary benzo[c]phenantridine alkaloids sanguinarine and chelerythrine. The alkaloid extract of M. cordata (MCE) is known for its antimicrobial and anti-inflammatory activities and thus is used as an active component in dental care products. Aim of work: The study examined the effects of MCE on lipopolysaccharide-induced inflammation and oxidative stress in human gingival fibroblasts. Material and Methods: The effect of MCE (0.01–10 μg/ml) on human gingival fibroblasts viability was evaluated using neutral red incorporation in the cells. Non-toxic concentrations of the extract (0.25 and 0.5 μg/ml) were used for evaluation of MCE effects on GSH level (GSH), reactive oxygen species (ROS) generation, cyclooxygenase-2 (COX-2) and interleukine-6 (IL-6) expression in cells treated with lipopolysaccharide (LPS). Results: MCE components induced the increase in intracellular GSH, decrease in ROS generation and reduction of the IL-6 and COX-6 expression in LPS-treated human gingival fibroblasts. Conclusion: The alkaloid extract of M. cordata suppresses the inflammation and oxidative stress in human gingival fibroblasts and it could be practically used in the treatment of periodontitis and alveolitis.
- Klíčová slova
- protizánětlivý účinek, antioxidační účinek, reaktivní kyslíkové sloučeniny, gluthathion, lipopolysacharid, cyklooxygenáza-2,
- MeSH
- antiflogistika terapeutické užití MeSH
- antioxidancia terapeutické užití MeSH
- cyklooxygenasa 2 imunologie MeSH
- fibroblasty imunologie účinky léků MeSH
- gingiva imunologie účinky léků MeSH
- glutathion farmakologie MeSH
- interleukin-6 imunologie MeSH
- léky rostlinné čínské farmakologie terapeutické užití MeSH
- lidé MeSH
- lipopolysacharidy imunologie MeSH
- oxidační stres imunologie účinky léků MeSH
- Papaveraceae chemie klasifikace MeSH
- reaktivní formy kyslíku imunologie škodlivé účinky toxicita MeSH
- rostlinné extrakty terapeutické užití MeSH
- stomatologický výzkum MeSH
- Check Tag
- lidé MeSH
BACKGROUND AIMS: Microvesicles (MV) shed from the plasma membrane of eukaryotic cells, including human embryonic stem cells (hESC), contain proteins, lipids and RNA and serve as mediators of cell-to-cell communication. However, they may also contain immunogenic membrane domains and infectious particles acquired from xenogenic components of the culture milieu. Therefore, MV represent a potential risk for clinical application of cell therapy. METHODS: We tested the ability of hESC and their most commonly used feeder cells, mouse embryonic fibroblasts (MEF), to produce MV. We found that hESC are potent producers of MV, whereas mitotically inactivated MEF do not produce any detectable MV. We therefore employed a combined proteomic approach to identify the molecules that constitute the major components of MV from hESC maintained in a standard culture setting with xenogenic feeder cells. RESULTS: In purified MV fractions, we identified a total of 22 proteins, including five unique protein species that are known to be highly expressed in invasive cancers and participate in cellular activation, metastasis and inhibition of apoptosis. Moreover, we found that hESC-derived MV contained the immunogenic agents apolipoprotein and transferrin, a source of Neu5Gc, as well as mouse retroviral Gag protein. CONCLUSIONS: These findings indicate that MV represent a mechanism by which hESC communicate; however, they also serve as potential carriers of immunogenic and pathogenic compounds acquired from environment. Our results highlight a potential danger regarding the use of hESC that have previously been exposed to animal proteins and cells.
- MeSH
- antigeny heterofilní imunologie MeSH
- antigeny nádorové imunologie metabolismus MeSH
- apolipoproteiny imunologie metabolismus MeSH
- buněčné linie MeSH
- elektronová mikroskopie MeSH
- embryonální kmenové buňky cytologie imunologie metabolismus MeSH
- fibroblasty cytologie imunologie metabolismus MeSH
- genové produkty gag imunologie metabolismus MeSH
- kokultivační techniky MeSH
- lidé MeSH
- mikropartikule imunologie metabolismus MeSH
- myši MeSH
- proteiny regulující apoptózu imunologie metabolismus MeSH
- proteomika MeSH
- riskování MeSH
- skot MeSH
- tandemová hmotnostní spektrometrie MeSH
- tkáňová terapie - dějiny škodlivé účinky MeSH
- transferin imunologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Airway remodelling, a central feature of asthma, is characterized by an alteration in the size, mass or number of tissue components which occur in and around the trachea, bronchi and bronchioles in the airways in response to injury and/or inflammation.The present review focuses on the most recent literature on airway remodelling and on the different drugs commonly used or potentially useful in the treatment of asthma with a particular attention to the studies conducted by our group in the last few years. RECENT FINDINGS: The interaction between the epithelium and mesenchymal elements such as fibroblasts is essential for normal airway repair. An abnormal response of this epithelial-mesenchymal trophic unit has been proposed to be central to the airway pathology and physiology characteristic of asthma. Current treatments may indirectly control airway remodelling through a reduction of inflammation but such a kind of approach is only in part successful. SUMMARY: The clear understanding of the events that take place during remodeling and the targeting of its specific components will be helpful in the development of novel therapies that might restore lung function.
Účel přehledu Remodelace dýchacích cest – klíčová vlastnost astmatu – je charakterizována změnou velikosti, objemu nebo počtu složek tkání, k níž dochází uvnitř a v okolí trachey, bronchů a bronchiolů v reakci na poškození a/nebo zánět. Tento přehled se soustřeďuje na nejnovější literaturu o remodelaci dýchacích cest a na léky běžně používané nebo potenciálně účinné v léčbě astmatu. Zvláštní pozornost je věnována studiím provedeným naší skupinou v průběhu několika posledních let. Nové poznatky Pro přirozenou reparaci dýchacích cest má zásadní význam interakce mezi epitelem a buňkami mezenchymu, například fibroblasty. Abnormální odpověď této epiteliálně‑mezenchymální trofické jednotky je považována za klíčovou pro patologii a patofyziologii astmatu. Současné léčebné postupy mohou nepřímo ovlivnit remodelaci pomocí tlumení zánětlivého procesu, ale tento postup je úspěšný jen zčásti. Souhrn Při vývoji nových léků, které by mohly přispět k obnově funkce plic, bude zapotřebí dokonalé porozumění dějům odehrávajícím se při remodelaci a cílené ovlivnění jejích specifických složek.
Revmatoidní artritida (RA) je chronické autoimunitní zánětlivé onemocnění, které vede k destrukci kloubů. V synoviální tkáni postiženého kloubu je pozorována zvýšená proliferace a hyperplazie buněk synoviální intimy, tvorba prozánětlivých cytokinů, chemokinů a akumulace imunokompetentních buněk. V posledních letech jsou v popředí zájmu při výzkumu patogeneze RA synoviální fibroblasty. Ty jsou aktivovány prozánětlivými cytokiny makrofágů a lymfocytů, nacož následně odpovídají tvorbou enzymů degradujících matrix, které vedou k progresivní destrukci kloubu. RA synoviální fibroblasty však vykazují invazivní potenciál i nezávisle na přítomnosti imunokompetentních buněk a prozánětlivých cytokinů. Autor se v tomto článku věnuje morfologii fibroblastů, jejich narušené apoptóze, interakci fibroblastů s imunokompetentními buňkami, aktivaci synoviálních fibroblastů nezávisle na cytokinech a účasti aktivovaných fibroblastů na destrukci kloubu během patogeneze RA.
Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease leading to inevitable joint destruction. An increased proliferation and hyperplasia of synovial lining cells in synovial tissue of affected joint is observed. Moreover, production of pro-inflammatory cytokines, chemokines, and accumulation of immunocompetent cells within the joint is also observed. Current research is specifically focused on the concept of synovial fibroblasts in the pathogenesis of RA. These cells are activated by pro-inflammatory cytokines secreted by macrophages and leukocytes, thereafter responding by an increased production of matrix degrading enzymes that lead to progressive joint destruction. RA synovial fibroblasts have also invasive potential independently on the presence of both immunocompetent cells and pro-inflammatory cytokines. In this overview, the author discusses morphology of synovial fibroblasts, altered apoptosis, and interaction of fibroblasts with immunocompetents cells. Furthermore, cytokine-independent activation of synovial fibroblasts, and the role of activated fibroblasts in the process of joint destruction within the pathogenesis of RA are also mentioned.
