Cytokinins comprise a group of phytohormones with an organ-specific mode of action. Although the mechanisms controlling the complex networks of cytokinin metabolism are partially known, the role of individual cytokinin types in the maintenance of cytokinin homeostasis remains unclear. Utilizing the overproduction of single-chain Fv antibodies selected for their ability to bind trans-zeatin riboside and targeted to the endoplasmic reticulum, we post-synthetically modulated cytokinin ribosides, the proposed transport forms of cytokinins. We observed asymmetric activity of cytokinin biosynthetic genes and cytokinin distribution in wild-type tobacco seedlings with higher cytokinin abundance in the root than in the shoot. Antibody-mediated modulation of cytokinin ribosides further enhanced the relative cytokinin abundance in the roots and induced cytokinin-related phenotypes in an organ-specific manner. The activity of cytokinin oxidase/dehydrogenase in the roots was strongly up-regulated in response to antibody-mediated formation of the cytokinin pool in the endoplasmic reticulum. However, we only detected a slight decrease in the root cytokinin levels. In contrast, a significant decrease of cytokinins occurred in the shoot. We suggest the roots as the main site of cytokinin biosynthesis in tobacco seedlings. Conversely, cytokinin levels in the shoot seem to depend largely on long-range transport of cytokinin ribosides from the root and their subsequent metabolic activation.

• MeSH
• cytokininy fyziologie   MeSH
• fenotyp * MeSH
• homeostáza * MeSH
• isopentenyladenosin analogy a deriváty   metabolismus   MeSH
• protilátky produkované rostlinami fyziologie   MeSH
• regulátory růstu rostlin fyziologie   MeSH
• semenáček fyziologie   MeSH
• tabák fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• antigeny nádorové MeSH
• imunologické techniky MeSH
• interferony MeSH
• kultivované buňky MeSH
• lymfokiny MeSH
• protilátky MeSH

• MeSH
• Bacillus anthracis imunologie   MeSH
• buněčná imunita MeSH
• dítě MeSH
• imunoglobuliny analýza   MeSH
• lidé MeSH
• nádory hrtanu imunologie   MeSH
• papilom imunologie   MeSH
• předškolní dítě MeSH
• T-lymfocyty imunologie   MeSH
• tvorba protilátek MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH

Alergický zánět zaujímá klíčové postavení v patofyziologii bronchiálního astmatu. Je charakterizován akumulací eozinofilních granulocytů v plicní tkáni, k níž dochází vlivem abnormálních prozánětlivých lokálních podmínek, které mohou být způsobeny průnikem alergenů epitelovými strukturami s narušenými bariérovými funkcemi. Prozánětlivé prostředí je charakterizováno i přítomností vzorů vnitřního poškození. Dochází k abnormální polarizaci T lymfocytů k převažující aktivitě subsetu Th2. T lymfocyty subsetu Th2 jednak modulují B lymfocyty k produkci protilátek IgE, jednak stimulují prostřednictvím cytokinů, včetně interleukinu 5 (IL‐5), lokální aktivaci a proliferaci eozinofilů. V těchto procesech jsou zapojeny i buňky vrozené imunity ILC‐2. Ztráta homeostatických regulací může vést k lokální tvorbě autoprotilátek, které reagují s biologicky aktivními látkami v granulích eozinofilů. Lokálně tak dochází k aktivaci komplementového systému, jehož působením je dále prohloubeno poškození plicní tkáně. V léčbě nemocných s bronchiálním astmatem jsou nově používána biologika, která cílí na IL‐5. Tato biologika jsou molekulárně rozdílná a jsou aplikována rozdílným způsobem. To by mělo být zohledněno při klinickém rozhodování o nasazení biologické léčby.

Allergic inflammation is the crucial component of the pathophysiology of bronchial asthma. It is characterized by the accumulation of eosinophils in lung tissue induced by the presence of an abnormal local proinflammatory microenvironment caused by the penetration of allergens through impaired epithelial surfaces. In addition, the proinflammatory environment is also characterized by the presence of damage patterns. The result is an abnormal polarization of T cells to the Th2 subset. T cells of the Th2 subset are modulating B cells to produce antibodies, including IgE class. Th2 T cells can stimulate local eosinophilopoiesis and activation of eosinophils via various cytokines, including IL‐5. Innate immunity cells, ILC‐2, are also participating in these complex mechanisms. The loss of homeostatic regulations can lead to the local production of autoantibodies reacting with the content of eosinophil cytoplasmatic granules, which could be followed by the activation of the complement system, substantially contributing to the tissue damage. Monoclonal antibodies targeting the critical component of allergic inflammation, IL‐5, are now an integral part of the therapy of patients with bronchial asthma. There are substantial differences both in molecular structure and the way of administration for these biologics. These differences should be considered when biological therapy is suggested.

• MeSH
• alergie farmakoterapie   patofyziologie   MeSH
• biologická terapie metody   MeSH
• bronchiální astma * farmakoterapie   patofyziologie   MeSH
• eozinofily MeSH
• humanizované monoklonální protilátky terapeutické užití   MeSH
• imunoglobuliny MeSH
• interleukin-5 * antagonisté a inhibitory   terapeutické užití   MeSH
• lidé MeSH
• mediátory zánětu MeSH
• zánět MeSH
• Check Tag
• lidé MeSH

Crohnova nemoc (CD) je multifaktoriální onemocnění, v jehož patogenezi dominuje poškozující zánět zprostředkovaný aktivitou abnormálně polarizovaných subsetů Th1 a Th17 T lymfocytů. V porovnání s jinými imunopatologickými onemocněními jsou možnosti léčebného ovlivnění biologickou terapií u nemocných s CD omezeny. S výjimkou vedolizumabu jsou zatím téměř výlučně postaveny na neutralizaci účinků tumoru nekrotizujícího faktoru α. Z mnoha dalších testovaných biologik byla prokázána klinická účinnost pro unikátní monoklonální protilátku ustekinumab, cílící na podjednotku p40 sdílenou cytokiny IL-12/IL-23. Interleukin-12 je nezbytný pro polarizaci T lymfocytů do subsetu Th1, IL-23 je klíčovým pro úplnou aktivaci a přežívání subsetu Th17 T lymfocytů. Imunomodulační zásah do drah IL-12/IL-23 ustekinumabem významně tlumí poškozující zánět. Klinická aplikace ustekinumabu při tom není spojena s rizikem omezení obranných a bariérových funkcí, jak tomu je u jiných testovaných biologik, např. cílících na IL-17/IL-17R. V klinických studiích je ověřována i účinnost biologik neutralizujících podjednotku p19, unikátní pro IL-23, v léčebném ovlivnění nemocných s CD, u kterých byla již prokázána klinická účinnost v léčbě nemocných jinými imunopatologiemi.

Crohn´s disease (CD) is a multifactorial illness, in which pathogenesis involving damage to the inflammatory response, mediated by abnormally polarized Th1 and Th17 subsets of T cell, is a prominent feature. CD treatment options employing biological therapies, predominantly based on antitumor necrosis factor α biologics, are limited compared with other immunological diseases. With the exception of vedolizumab among numerous other biologics tested so far, clinical efficacy has recently been approved only for ustekinumab, which targets subunit p40 shared by both IL-12 and IL-23 interleukins. Interleukin-12 is indispensable for the functional polarization of T cells into the Th1 subset. Interleukin-23 is essential for the full activation and survival of the Th17 subset of T cells. Immunomodulatory intervention employing targeting of IL-12/IL-23 signaling pathways via ustekinumab alleviates inflammatory reactions in patients with CD. In addition, ustekinumab treatment is not associated with an increased risk of immunity impairment or decreased protective and barrier function of gut mucosa, which are common side effects of other biologics, especially those targeting IL-17/IL-17R signaling pathways. The clinical efficacy of another treatment targeting the unique subunit of p19 of IL-23 in CD patients is now being tested in ongoing clinical trials.

• MeSH
• biologická terapie MeSH
• buňky Th17 fyziologie   účinky léků   MeSH
• Crohnova nemoc * diagnóza   farmakoterapie   patofyziologie   MeSH
• interleukin-12 MeSH
• interleukin-23 MeSH
• lidé MeSH
• monoklonální protilátky MeSH
• signální transdukce * MeSH
• střevní mikroflóra MeSH
• Th1 buňky fyziologie   účinky léků   MeSH
• ustekinumab * terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) can selectively trigger apoptosis in various cancer cell types. However, many cancer cells are resistant to death receptor-mediated apoptosis. Combination therapy with platinum complexes may affect TRAIL-induced signaling via modulation of various steps in apoptotic pathways. Here, we show that cisplatin or a more potent platinum(IV) complex LA-12 used in 20-fold lower concentration enhanced killing effects of TRAIL in human colon and prostate cancer cell lines via stimulation of caspase activity and overall apoptosis. Both platinum complexes increased DR5 surface expression in colon cancer cells. Small interfering RNA-mediated DR5 silencing rescued cells from sensitizing effects of platinum drugs on TRAIL-induced caspase-8 activation and apoptosis, showing the functional importance of DR5 in the effects observed. In addition, both cisplatin and LA-12 triggered the relocalization of DR4 and DR5 receptors to lipid rafts and accelerated internalization of TRAIL, which may also affect TRAIL signaling. Collectively, modulations of the initial steps of the extrinsic apoptotic pathway at the level of DR5 and plasma membrane are important for sensitization of colon and prostate cancer cells to TRAIL-induced apoptosis mediated by LA-12 and cisplatin.

• MeSH
• amantadin * analogy a deriváty   farmakologie   MeSH
• apoptóza * fyziologie   účinky léků   MeSH
• cisplatina * farmakologie   MeSH
• fluorescenční protilátková technika MeSH
• konfokální mikroskopie MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory * metabolismus   MeSH
• organoplatinové sloučeniny * farmakologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protein TRAIL * metabolismus   MeSH
• průtoková cytometrie MeSH
• RNA interference MeSH
• separace buněk MeSH
• signální transdukce fyziologie   účinky léků   MeSH
• TRAIL receptory metabolismus   MeSH
• transport proteinů účinky léků   MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Lactococcus lactis, a probiotic bacterium of food origin, has recently been demonstrated as a suitable strain for the production and in vivo delivery of therapeutically important proteins into the gut. We aimed to engineer recombinant L. lactis cells producing/secreting REX binding proteins that have been described as IL-23 receptor (IL-23R) blockers and IL-23R antagonists suppressing the secretion of cytokine IL-17A, a pivotal step in the T-helper Th17-mediated pro-inflammatory cascade, as well as in the development of autoimmune diseases, including inflammatory bowel disease (IBD). To reach this goal, we introduced cDNA sequences coding for REX009, REX115, and REX125 proteins into plasmid vectors carrying a Usp45 secretion signal, a FLAG tag sequence consensus, and a LysM-containing cA surface anchor (AcmA), thus allowing cell-surface peptidoglycan anchoring. These plasmids, or their non-FLAG/non-AcmA versions, were introduced into L. lactis host cells, thus generating unique recombinant L. lactis-REX strains. We demonstrate that all three REX proteins are expressed in L. lactis cells and are efficiently displayed on the bacterial surface, as tested by flow cytometry using an anti-FLAG antibody conjugate. Upon 10-fold concentration of the conditioned media, a REX125 secretory variant can be detected by Western blotting. To confirm that the FLAG/non-FLAG REX proteins displayed by L. lactis retain their binding specificity, cell-surface interactions of REX proteins with an IL-23R-IgG chimera were demonstrated by flow cytometry. In addition, statistically significant binding of secreted REX009 and REX115 proteins to bacterially produced, soluble human IL-23R was confirmed by ELISA. We conclude that REX-secreting L. lactis strains were engineered that might serve as IL-23/IL-23R blockers in an experimentally induced mouse model of colitis.

• Publikační typ
• časopisecké články MeSH

Members of neuropeptide B/W signaling system have been predominantly detected and mapped within the CNS. In the rat, this system includes neuropeptide B (NPB), neuropeptide W (NPW) and their specific receptor NPBWR1. This signaling system has a wide spectrum of functions including a role in modulation of inflammatory pain and neuroendocrine functions. Expression of NPB, NPW and NPBWR1 in separate heart compartments, dorsal root ganglia (DRG) and stellate ganglia was proven by RT-qPCR, Western blot (WB) and immunofluorescence. Presence of mRNA for all tested genes was detected within all heart compartments and ganglia. The presence of proteins preproNPB, preproNPW and NPBWR1 was confirmed in all the chambers of heart by WB. Expression of preproNPW and preproNPB was proven in cardiac ganglionic cells obtained by laser capture microdissection. In immunofluorescence analysis, NPB immunoreactivity was detected in nerve fibers, some nerve cell bodies and smooth muscle within heart and both ganglia. NPW immunoreactivity was present in the nerve cell bodies and nerve fibers of heart ganglia. Weak nonhomogenous staining of cardiomyocytes was present within heart ventricles. NPBWR1 immunoreactivity was detected on cardiomyocytes and some nerve fibers. We confirmed the presence of NPB/W signaling system in heart, DRG and stellate ganglia by proteomic and genomic analyses.

• MeSH
• exprese genu MeSH
• fluorescenční protilátková technika MeSH
• ganglion stellatum metabolismus   MeSH
• myokard metabolismus   MeSH
• neuropeptidy genetika   imunologie   metabolismus   MeSH
• potkani Zucker MeSH
• receptory neuropeptidů genetika   imunologie   metabolismus   MeSH
• receptory spřažené s G-proteiny genetika   imunologie   metabolismus   MeSH
• reprodukovatelnost výsledků MeSH
• signální transdukce MeSH
• spinální ganglia metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

This review describes the IL-20 family of cytokines in rheumatoid arthritis (RA) and spondyloartrhitits (SpA) including psoriatic arthritis. The IL-20 receptor (R) cytokines IL-19, IL-20, and IL-24 are produced in both the peripheral blood and the synovial joint and are induced by Toll-like receptor ligands and autoantibody-associated immune complexes in monocytes. IL-19 seems to have anti-inflammatory functions in arthritis. In contrast, IL-20 and IL-24 increase the production of proinflammatory molecules such as monocyte chemoattractant protein 1 and are associated with bone degradation and radiographic progression. IL-22 is also associated with progression of bone erosions. This suggests that the IL-22RA1 subunit shared by IL-20, IL-22, and IL-24 is important for bone homeostasis. In line with this, the IL-22RA1 has been found on preosteoclasts in early RA. IL-26 is produced in high amounts by myofibroblasts and IL-26 stimulation of monocytes is an important inducer of Th17 cells in RA. This indicates a role for IL-26 as an important factor in the interactions between resident synovial cells and infiltrating leukocytes. Clinical trials that investigate inhibitors of IL-20 (fletikumab) and IL-22 (fezakinumab) in psoriasis and RA have been terminated. Instead, it seems that the strategy for modulating the IL-20 cytokine family should take the overlap in cellular sources and effector mechanisms into account. The redundancy encourages inhibition of more than one cytokine or one of the shared receptors. All IL-20 family members utilize the Janus kinase signaling pathway and are therefore potentially inhibited by drugs targeting these enzymes. Effects and adverse effects in ongoing clinical trials with inhibitors of IL-22 and the IL-22RA1 subunit and recombinant IL-22 fusion proteins will possibly provide important information about the IL-20 subfamily of cytokines in the future.

• MeSH
• humanizované monoklonální protilátky MeSH
• interleukiny antagonisté a inhibitory   imunologie   metabolismus   MeSH
• Janus kinasy imunologie   metabolismus   MeSH
• klinické zkoušky jako téma MeSH
• lidé MeSH
• monoklonální protilátky farmakologie   terapeutické užití   MeSH
• neutralizující protilátky farmakologie   terapeutické užití   MeSH
• osteoklasty imunologie   metabolismus   MeSH
• psoriatická artritida farmakoterapie   imunologie   MeSH
• receptory interleukinů antagonisté a inhibitory   imunologie   metabolismus   MeSH
• revmatoidní artritida farmakoterapie   imunologie   MeSH
• signální transdukce imunologie   MeSH
• široce neutralizující protilátky MeSH
• synoviální membrána cytologie   imunologie   metabolismus   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Size control is a fundamental question in biology, showing incremental complexity in plants, whose cells possess a rigid cell wall. The phytohormone auxin is a vital growth regulator with central importance for differential growth control. Our results indicate that auxin-reliant growth programs affect the molecular complexity of xyloglucans, the major type of cell wall hemicellulose in eudicots. Auxin-dependent induction and repression of growth coincide with reduced and enhanced molecular complexity of xyloglucans, respectively. In agreement with a proposed function in growth control, genetic interference with xyloglucan side decorations distinctly modulates auxin-dependent differential growth rates. Our work proposes that auxin-dependent growth programs have a spatially defined effect on xyloglucan's molecular structure, which in turn affects cell wall mechanics and specifies differential, gravitropic hypocotyl growth.

• MeSH
• Arabidopsis fyziologie   MeSH
• buněčná stěna metabolismus   MeSH
• fluorescenční protilátková technika MeSH
• fyziologie rostlin * MeSH
• glukany chemie   metabolismus   MeSH
• hrách setý fyziologie   MeSH
• kyseliny indoloctové metabolismus   MeSH
• regulace genové exprese u rostlin MeSH
• rostlinné buňky metabolismus   MeSH
• signální transdukce MeSH
• vývoj rostlin * MeSH
• xylany chemie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH