S78 Abstracts monitored by measuring the proteoglycan release into the supernatant (DMB-assay) and by Safranin-O-staining of histological sections. Total MMP-activity in the supernatant was quantifi ed using a fl uorescence assay with a broadly MMP-specifi c peptide substrate. Classifi cation of the individual MMP‘s was achieved by gelatine zymography. Morphology of SF and cartilage surface was examined by raster electron microscopy (REM). Results:For all stimuli proteoglycan release following co-culture of BC with RA-, OA- and JT-SF was quantitatively higher than in the corresponding BC-monoculture. In BC-monoculture stimulation with IL-1β or IL-1β/TNF-α, but not TNF-α alone, signifi cantly increased proteoglycan depletion as compared to non stimulated controls. In co-culture with RA-, OA-, or JT-SF, however all stimulations led to signifi cantly higher proteoglycan release compared to non-stimulated controls. Interestingly, signifi cantly higher levels of total MMP-activity were observed for all stimuli in both BC-monoculture and co-culture with RA-, OA- and JT- SF. Quantitatively however, the total MMP-activity in stimulated co-cultures showed up to 4 fold higher levels compared with BC-monoculture. In zymography cytokine treated BC-monocultures and co-cultures with SF showed an increased MMP-2 and MMP-9 activity, though co-cultures led to distinct higher values than BC-monoculture. REM revealed that cytokine treatment induced: i) degradation of the cartilage matrix already in BC-monoculture; and ii) an activated phenotype of SF in co-culture. Conclusion: Stimulation with TNF-α, IL-1β or TNF-α/IL-1β results in increased cartilage degradation and MMP-activity in both BC-monoculture and co-culture with SF. Signifi cantly or numerically increased overall MMP-, MMP-2-, and MMP-9 activity following co-culture with SF, on the other hand, underlines the central role of SF for cartilage degradation. Comparison of the 2 culture systems may allow to distinguish the relative contribution of BC and SF to cartilage degradation. POFER-12 Striktly time-dependent regulation of the mRNA expression of pro-infl ammatory/pro-destructive genes in sfb by TNF-alpha Kunisch E., Gandesiri M., Lux S., Jansen A., Kinne RW. Nachwuchsgruppe Experimentelle <strong>Rheumatologie</strong>, Friedrich-Schiller-Universität Jena Purpose: TNF-alpha is a major inductor of pro-destructive/pro-infl ammatory processes in rheumatoid arthritis (RA), whose infl uence on RA synovial fi broblast (SFB) functions has been extensively studied. However, there are limited data about the time-dependent induction of these functions by TNF-alpha. Th erefore, the present study sought to characterize the time-dependent induction of mRNA expression for MMP-1, MMP-3, COX2, IL-8 and IL-6 in RA-, osteoarthritic (OA)-, and joint trauma (JT)-SFB following TNF-alpha stimulation. Methods: For analysis of time-dependent mRNA expression for MMP- 1, MMP-3, COX2, IL-8, and IL-6, RA-, OA-, and JT-SFB (beginning of 3rd passage) were stimulated with TNF-alpha (10 ng/ml) for 0, 1, 2, 4, 6, 8, 10, and 24 h. mRNA expression was analyzed by real-time RT-PCR. Results: COX2 and IL-6 mRNA reached their maxima 1h aft er TNFalpha stimulation, with a strong decrease until about 8 h and a slight increase thereaft er. IL-8 mRNA was also induced aft er 1 h stimulation but had a maximum following 4<strong>–</strong>8 h TNF-alpha stimulation. In contrast, MMP-1 and MMP-3 mRNA expression showed a continuous increase in RA-, OA-, and JT-SFB over the 24 h period of TNF-alpha stimulation. Conclusions: Time-dependent induction of mRNA for the pro-destructive/pro-infl ammatory molecules MMP-1, MMP3, COX2, IL-8 and IL-6 was observed in RA-, OA-, and JT-SFB following TNF-alpha stimulation. Whereas pro-infl ammatory molecules showed their maximal mRNA expression as early as 1h, pro-destructive molecules did not reach their maxima until 24 h aft er TNF-alpha stimulation. Th ese data show a strictly time-dependent, possibly cascade-signal regulated mRNA expression for pro-destructive/pro-infl ammatory molecules in | <strong>Zeitschrift</strong> <strong>für</strong> <strong>Rheumatologie</strong> · <strong>Supplement</strong> 1 · 2006 RA, OA-, and JT-SFB following TNF-alpha stimulation with potential relevance for pathophysiological and therapeutic aspects. Th is study was supported by the German Federal Ministry of Education and Research (BMBF; grant FKZ 01ZZ0105 to R.W. Kinne, Interdisciplinary Center for Clinical Research Jena) and the German Research Foundation (DFG; grant KI 439/7-1 to R.W. Kinne), as well as a grant for the advancement of female scientists to E. Kunisch (LUBOM Th uringia 05/2005-05/2005). POFER-13 Low baseline serum cortisol predicts marked clinical improvement 7 days after initiation of anti-TNF antibody therapy in rheumatoid arthritis Straub RH. 1 , Härle P. 1 , Pongratz G. 1 , Fleck M. 1 , Cutolo M. 2 , Atzeni F. 3 , Antoni C. 4 , Kalden JR. 5 , Lorenz HM. 6 , Sarzi-Puttini P. 3 1 Laboratory of Exp. Rheumatology and Neuroendocrino-Immunology, Dept. of Internal Medicine I, University Hospital Regensburg, 93042 Regensburg, Germany, 2 Division of Rheumatology, Department of Internal Medicine and Medical Specialties, University of Genova, Italy, 3 Rheumatology Unit, University Hospital L Sacco, Milan, Italy, 4 Schering-Plough Research Institute, Kenilworth, NJ 07033, U.S.A., 5 Dept. of Internal Medicine III, University of Erlangen<strong>–</strong>Nürnberg, 91054 Erlangen, Germany, 6 Division of Rheumatology, Dept. of Internal Medicine V, University of Heidelberg, 69120 Heidelberg, Germany Objective: It is known clinical experience that some patients with rheumatoid arthritis (RA) rapidly profi t from anti-TNF antibody therapy whereas others show no immediate benefi t. Th is investigation studied the predictive role of hypothalamic <strong>–</strong> pituitary <strong>–</strong> adrenal (HPA) axis hormones for immediate clinical improve-ment during anti-TNF antibody therapy. Methods: In this study in 24 RA patients, we measured at baseline adrenocorticotropic hormone (ACTH), 17-hydroxyprogesterone (17OHP), cortisol, and interleukin (IL)-6. Immediate clinical improve-ment was judged 7 days aft er initiation of therapy by the physician global estimate of disease activity, which appraises pain, well-being, and functional ability. Results: Compared to patients with little improvement, patients with >50% improvement were not diff erent in age, gender, accompanying therapies, and <strong>–</strong> at baseline <strong>–</strong> physician global assessment, ESR, serum IL-6 and ACTH. However, patients with > 50% improvement had lower baseline serum levels of cortisol and a lower baseline ratio of serum cortisol divided by serum 17OHP. Th e extent of improvement negatively correlated with baseline serum cortisol (R=-0.457, p=0.025) and the cortisol/17OHP ratio (R=-0.589, p=0.003). In the longitudinal part of this study over 12 weeks, those patients with >50% improve-ment demonstrated steadily increasing serum levels of cortisol, which was not observed in patients with little improvement. Conclusions: Since TNF inhibits adrenal conversion of 17OHP into cortisol leading to low serum corti-sol, these fi ndings indicate that some patients rapidly profi t from TNF neutralization probably by restoring this important enzyme step. In responders, TNF neutralization leads to an increase in serum cortisol and rapid clinical improvement.
POFER-14 Serum cartilage oligomeric matrix protein (SCOMP): comparison of patients with psoriatic arthritis (PSA) and spondylitis ankylosans (SPA) Haberhauer G. 1 , Skoumal M. 2 , Fink A. 3 , Steiner A. 3 , Wottawa A. 1 1 Institute for Rheumatology of the Kurstadt Baden in Cooperation with the Danube-University Krems, Austria, 2 Ludwig Boltzmann Institute of Osteology at the Hanusch-Hospital of WGKK and AUVA Trauma Centre Meidling,4 th Medical Dpt., 3 Dept. Of Dermatology, Wilhelminen-Hospital Vienna, Austria. Background: COMP is a pentameric protein of fi ve identical disulfi delinked subunits and belongs to the thrombospondin family of proteins. It is proposed to be a marker of cartilage degredation in patients with rheumatoid arthritis and osteoarthritis. Objective: Th e aim of this study was to prove serum COMP (sCOMP) as a parameter for disease activity in patients with active PsA and SPA with peripheral joint involvement. Methods: Serum levels of COMP were measured in 64 patients with PsA and psoriasis vulgaris (m/f: 39/25) and 38 patients with SPA (m/f:30/8). Active PsA was defi ned by a minimum of 2 swollen joints and serum CRP levels >20mg/dl. Th is could be detected in 25 PsA patients. Seven SPA patients showed peripheral arthritis.Routine laboratory monitoring and clinical assessment of the disease status (TJC,SJC) were performed additionally. SCOMP was measured by a commercially available sandwich-type ELISA-kit developed by AnaMar Medical AB, Sweden.Statistical evaluation was calculated by general linear models procedure, Ryan-Einot-Gabriel-Welsch Multiple F test. Results: In our 68 patients with PsA we could fi nd sCOMP levels from 6.7<strong>–</strong>33,8 U/l (mean: 12,7), and in the SPA patients from 3.9<strong>–</strong>15.1 U/l (mean:9,4). Patients with active PsA had sCOMP levels from 10.9<strong>–</strong>33.0 U/l (mean: 17,7) and SPA patients with peripheral arthritis from 9.5<strong>–</strong>15.1 U/l (mean: 12.5). Serum COMP levels of PsA patients were signifi cantly higher compared to the SpA patients (p
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