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  • 1
    ISSN: 1600-0625
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Whereas the effects of interleukin (IL)-10 on several epithelial cell types are well established the capability of IL-10 to target keratinocytes (KC) is still a matter of debate. This, however, is of considerable importance, as IL-10 is a major anti-inflammatory, immunosuppressive cytokine with impact on the cutaneous homeostasis. Recently, IL-10 therapy has been proven to be clinically effective in psoriasis. Response to therapy is associated with normalization of typical parameters of keratinocyte pathology, but it is unclear whether this results from direct or indirect (secondary) effects. The purpose of the present study was to further investigate direct effects of IL-10 on keratinocytes and to address the reason for potential IL-10 unresponsiveness using keratinocytes such as the cell line HaCaT as well as primary foreskin keratinocytes. Using real time RT-PCR we demonstrated that IL-10 is neither able to induce its typical early gene product suppressor of cytokine signalling (SOCS) 3 nor to modulate the interferone (IFN)-γ-induced expression of SOCS 1 and 3. Although flow cytometric analyses showed binding of biotin labelled IL-10 to HaCaT cells, blocking experiments indicated that this resulted from unspecific binding. Moreover, scatchard plot analyses excluded specific binding to primary KC and HaCaT cells. Finally, real time mRNA analyses and Western blot experiments demonstrated that the absence of any specific binding results from the absence of clear IL-10R1 (alpha chain) expression, whereas the IL-10R2 (beta chain) is strongly expressed. Our data indicates that IL-10 unresponsiveness of keratinocytes could be explained by the lacking of functional IL-10 receptor expression and suggest that any IL-10 effects on these cells observed are indirectly mediated.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature America Inc.
    Nature biotechnology 17 (1999), S. 271-275 
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] We synthetically reconstructed a discontinuous binding site on interleukin-10 (IL-10) that recognizes the neutralizing anti–IL-10 antibody CB/RS/1. To design the 32-mer IL-10 mimic, a discontinuous interaction site on IL-10 was mapped, and binding studies with epitope-derived peptides led ...
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1173
    Keywords: Schlüsselwörter IL-10 ; Zytokine ; Inflammation ; Dermatitis ; Psoriasis ; Key words IL-10 ; Cytokines ; Inflammation ; Psoriasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary In recent years the investigation of cytokines has been a focus of scientific interest in order to understand the pathogenesis of a variety of diseases. In additions cytokines are increasingly being used for therapeutic purposes, including dermatologic disorders. IL-10 is a recently described cytokine with anti- inflammatory and immunosuppressive qualities which plays an important role in immunoregulation. The overexpression of this mediator has been proven in some inflammatory dermatoses as well as in various skin tumors. These observations help to understand down regulatory events in hyperinflammatory conditions such as allergic or toxic dermatitis on the one hand and the suppression of an adequate anti-tumor response and thereby the progression of malignant tumors on the other hand. Recent investigations indicate a relative IL-10 deficiency in psoriasis. Initial therapeutic applications of IL-10 in psoriasis underline the pathophysiological importance of this cytokine.
    Notes: Zusammenfassung Seit einiger Zeit steht der Nachweis von Zytokinen auch in der Dermatologie im Mittelpunkt des wissenschaftlichen Interesses, insbesondere da diese ein besseres Verständnis der Pathogenese verschiedenster Erkrankungen erlauben. Darüber hinaus finden Zytokine zunehmend auch in der Dermatologie therapeutische Anwendung. Interleukin-(IL-) 10 ist ein erst seit kurzem bekanntes Zytokin, welches in der Immunregulation insbesondere durch seine antiinflammatorischen und immunsuppressiven Eigenschaften eine herausragende Rolle besitzt. Zwischenzeitlich konnte die verstärkte Expression dieses Mediators bei einigen entzündlichen Dermatosen ebenso wie bei verschiedenen Malignomen der Haut nachgewiesen werden. Diese Beobachtungen sind von Bedeutung, da sie einerseits die gewünschte Limitierung von hyperinflammatorischen Prozessen wie bei Ekzemen und Erythemen, andererseits aber auch die Unterdrückung einer adäquaten anti-Tumorantwort und somit Progredienz von Malignomen erklären könnten. Neuere Untersuchungen zeigen, daß bei der Psoriasis eine relative IL-10-Defizienz besteht. Erste therapeutische Applikationen von IL-10, die eine antipsoriatische Potenz vermuten lassen, sprechen auch hier für die pathophysiologische Bedeutung dieses Zytokins.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-501X
    Keywords: TNF ; IL-10 ; Discontinuous epitope ; Antibody ; Peptide scan ; Protein-protein interaction ; Spot synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary We have characterized the interaction of two monoclonal antibodies with their respective antigens using cellulose-bound sets of overlapping peptides (peptide scans). Both antibodies CB/RS/5 and CB/MT/1 recognize discontinuous epitopes present in human interleukin-10 (IL-10) and tumor necrosis factor alpha (TNF-α). In addition, the interaction between TNF-a and its 55-kDa receptor (TNF-R) was investigated by the same approach. Both antibodies, as well as TNF-α, interacted with two or more regions of the peptide scans. Antibody-binding competition studies between the native antigens and peptides, covering single parts of the binding regions, enabled us to distinguish between binding to the paratope or other regions of the antibody. The combination of these experimental approaches allowed the identification of short antigen-derived sequences that are separated on the primary sequence but close in space on the surface of IL-10 and TNF-α, thus representing putative discontinuous epitopes. In the case of the TNF-R-derived peptide scans, two of the identified regions interact with the structurally similar TNF-β in the TNF-β-TNF-R complex. These data indicate that this approach should be generally applicable for mapping nonlinear protein-protein contact sites.
    Type of Medium: Electronic Resource
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