Publikationsdatenbank

Promoter squelching as a tool to downregulate the inflammatory cascade in canine osteoarthritis (2006)

Art

Poster

Autoren

Rachakonda, S.

Rai, M. F.

Manning, K. S.

Schmidt, M. F. G.

Kongress

6th World Congress of the International Cartilage Repair Society

Prague/ Chech Republic, 07. – 10.12.2006

Quelle

Osteoarthritis and cartilage

Bandzählung: 14, Suppl. B

Seiten: 135

ISSN: 1063-4584

Sprache

Englisch

Kontakt

Institut für Immunologie

Robert-von-Ostertag-Str. 7-13
Gebäude 35
14163 Berlin
+49 30 838 51834

Abstract / Zusammenfassung

P249<br><br

Purpose:<br>Osteoarthritis (OA) characterised by cartilage degradation
is a severe joint disorder affecting animals and humans
alike. Canine species provide a useful model to study this disease
process as it mimics the human form of OA both clinically
and pathophysiologically. It has been proven that IL-1β and TNFα
are the inflammatory cytokines that accentuate the disease process.
The absence of Th2 cells in the inflammatory synovial fluid
points out that this disorder could be corrected by installing a
balance between the Th1 and Th2-response. Previous work has
shown that Interleukin 4 (IL-4) downregulates the inflammatory
pathway and therefore is a good candidate to combat OA in
canine species.<br><br
Methods:<br>Present work focuses on simulating the OA disease
process in-vitro using canine chondrocytes in monolayer and 3D
cell culture systems. Our approach is based on dosage regulated
IL-4 expression under cell culture conditions. To achieve this we
are using promoter squelching as a method to self regulate
IL-4 expression involving the canine COX-2 promoter which we
partially sequenced.
Results: Our experiments have shown that canine chondrocytes
can be passaged till the 7th generation without any change in
morphology. We could also show by qPCR that these cells over
express inflammatory cytokines such as IL-6, IL-8 and IL-18 in
comparison to the house keeping gene GAPDH upon exogenous
stimulation with either human recombinant IL-1β or TNFα.<br><br
Conclusions: <br>We report on the optimization of our canine invitro
model for OA therapy which includes IL-1β and TNFα
synthesis from canine PBMC as well as a reporter assay for
COX-2 promoter activity for our indigenously designed vector.
Efforts are on the way to increase expression levels of canine
IL-4 in cell culture.