Quantification of cytokines and inflammatory mediators in a three-dimensional model of inflammatory arthritis.
Human recombinant IL-1beta and TNFalpha have been previously used to induce a cytokine response in canine chondrocytes. In order to establish this functional relation in a homologous system in vitro, we have developed both 2D and 3D models of inflammatory arthritis using canine recombinant cytokines in canine articular chondrocytes. IL-1beta and TNFalpha were cloned and subsequently expressed in Escherichia coli. The purified recombinant canine cytokines were used to simulate inflammation in vitro and the expression of typical inflammation markers such as proinflammatory cytokines (IL-1beta, IL-6, IL-8, GM-CSF and TNFalpha), enzyme mediators (MMP-3 MMP-13, iNOS, COX-2) and their catabolites (NO, PGE(2)) was measured. High expression of proinflammatory cytokines, enzyme mediators and their catabolites was only observed in IL-1beta/TNFalpha stimulated cells. We conclude that the canine IL-1beta and TNFalpha generated in this study are biologically active and equally effective in the canine cell culture systems. Inducing an inflammatory pathway by canine exogenous cytokines in canine chondrocytes provides a useful tool for the study of canine inflammatory arthritis.
Rai
Muhammad F
MF
Institute of Immunology and Molecular Biology, Faculty of Veterinary Medicine, Free University Berlin, Philippstrasse 13, D-10115 Berlin, Germany.
Rachakonda
P Sivaramakrishna
PS
Manning
Kizzie
K
Vorwerk
Brita
B
Brunnberg
Leo
L
Kohn
Barbara
B
Schmidt
Michael F G
MF
D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000276:immunology; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000276:immunology*; D015232:Dinoprostone / Q000378:metabolism; D004195:Disease Models, Animal; D004285:Dogs; D006801:Humans; D018836:Inflammation Mediators / Q000276:immunology*; D053583:Interleukin-1beta / Q000235:genetics / Q000276:immunology*; D053509:Matrix Metalloproteinase 13 / Q000235:genetics / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000235:genetics / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D010003:Osteoarthritis / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology*
D053674:Aggrecans; D024043:Collagen Type II; D016207:Cytokines; D018836:Inflammation Mediators; D053583:Interleukin-1beta; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3; D015232:Dinoprostone