Dokument: Die Expression und funktionelle Rolle von GranzymB in polymorphkernigen neutrophilen Granulozyten bei der Rheumatoiden Arthritis
| Titel: | Die Expression und funktionelle Rolle von GranzymB in polymorphkernigen neutrophilen Granulozyten bei der Rheumatoiden Arthritis | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=17279 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20110228-115115-5 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Deutsch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten | |||||||
| Medientyp: | Text | |||||||
| Autor: | Bartsch, Bianca [Autor] | |||||||
| Dateien: |
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| Dewey Dezimal-Klassifikation: | 600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit | |||||||
| Beschreibung: | Ch. Iking-Konert, B. Bartsch, O. Sander, B. Ostendorf, Ch. Specker, M.G. Hänsch, M. Schneider
Granzyme B is transcribed and expressed in PMN in synovial fluid of patients with rheumatoid arthritis Objectives: Granzyme B (GrB) is a serine protease usually described in cytotoxic T-cells and natural killer cells with a potential role in cartilage and joint destruction in RA (Froelich 1993, Tak 1999, Rondey 2001). We described, that GrB is also functionally expressed in polymorphonuclear neutrophils (PMN) in vitro (Wagner 2004). The contribution of PMN in tissue destruction in RA appears to be obvious, based on the fact, that prominent PMN infiltrates are found in inflamed joints. How they might contribute to the tissue destruction however is not well characterised. For that we studied, whether GranB is transcribed and expressed in PMN in vivo in patients with active RA. Material and Methods: GrB expression was measured intracellular and extracellular in highly purified PMN of the synovial fluid (SF) of patients with RA (n =20) by FACScan, confocal laser microscopy and with PCR as well. Additionally the GrB expression in PMN was measured in whole blood. As a control group patients with arthritis other than RA were assessed (n = 10). The quantity of GrB in the SF was measured by ELISA, as was COMP, CCP, rheumatoid factor. The GrB expression in PMN was compared with clinical and serological parameters such as DAS28, CRP and CCP-status. The expression of GrB in vitro was studied after stimulation of PMN populations from whole blood as well as synovial fluid, with TNFα, IL2, IL15 and γIFN. Results: PMN in the SF of patients with active RA transcribe GrB, by PCR GrB specific RNA could be detected in highly, CD15 purified PMN. By specific antibodies against GrB, the protein was found by confocal laser microscopy as well as by FASCcan analysis in PMN. In whole blood of patients with RA (n=9), but less in the SF, GrB is also expressed on the surface of PMN. By ELISA on the other hand significant amounts of GrB were found in the SF. When compared to patients with non erosive arthritis (n = 10) we found a significant lower GrB expression intracellular PMN (48,5 +/- 40,1% vs. 95 +/- 2,4 % positive PMN; p = 0,017), but more soluble GrB in the SF (241,4 +/- 475,9 pg/ml vs. 48,9 +/- 71,3 pg/ml; p 0,09). No correlation between the amount of GrB in the SF or in PMN with CRP, DAS28 or CCP was detected. TNFα, γIFN, as well as IL2, but most prominent IL15 was able to decrease the amount of GrB in PMN. When isolated PMN were stimulated with IL15, GrB could be detected in the supernatant, suggesting a release of GrB from PMN by IL15. Discussion: PMN in the SF of patients with RA express GrB. This is the first report of GrB-positive PMN in vivo. The release and functional activation of GrB in PMN might be induced by proinflammatory cytokines such as TNFa and IL 15, both playing a crucial role in the pathogenesis of RA. GrB levels were described as an independent predictor for the development of erosions by others earlier, but the phenotype of up to 50% of GrB positive cells in the SF could not be identified. Our data show, that PMN might be missed, as specific PMN surface markers were not used. Traditionally PMN are thought to play a dominant role in the host defence, e.g. against bacterial infections. Nevertheless, by the release of cytotoxic and proteolytic entities, including GrB, PMN might also contribute to chronic inflammatory processes, e.g. to the destruction of cartilage and bone in RA. | |||||||
| Lizenz: |  Urheberrechtsschutz | |||||||
| Fachbereich / Einrichtung: | Medizinische Fakultät | |||||||
| Dokument erstellt am: | 28.02.2011 | |||||||
| Dateien geändert am: | 28.02.2011 |
