Zeitschrift für Rheumatologie – Supplement 1 - Deutsche ...

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 Rheumatologie, 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 4–8 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
| Zeitschrift für Rheumatologie · Supplement 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–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 – pituitary – 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 – at baseline – 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.