The effect of interleukin-1 on cytokine gene expression in cultured human articular chondrocytes analyzed by messenger RNA phenotyping.
OBJECTIVE
To investigate the pattern of cytokine gene expression in human articular chondrocytes in culture in response to interleukin-1 beta (IL-1 beta). The effect of serum and variations in culture conditions was also studied.


METHODS
Messenger RNA was extracted from cells, reverse-transcribed to complementary DNA, and amplified by the polymerase chain reaction (PCR), using specific oligonucleotide primers. The PCR products were validated by restriction analysis with specific enzymes and by Southern blot analysis.


RESULTS
In cultured articular chondrocytes, IL-1 beta, IL-1 alpha, granulocyte colony-stimulating factor (CSF), and granulocyte-macrophage CSF cytokine genes were expressed only after induction by IL-1 beta. However, IL-6, IL-8, and macrophage CSF genes were expressed constitutively. The expression of IL-1 beta was dose and time dependent.


CONCLUSIONS
Using PCR, it was possible to demonstrate gene expression for several cytokines in human articular chondrocytes in culture. It was evident that some cytokine genes were expressed constitutively and some were inducible by IL-1 beta.
Seid
J M
JM
Department of Human Metabolism and Clinical Biochemistry, Sheffield University Medical School, United Kingdom.
Rahman
S
S
Graveley
R
R
Bunning
R A
RA
Nordmann
R
R
Wishart
W
W
Russell
R G
RG
D001483:Base Sequence; D015139:Blotting, Southern; D002358:Cartilage, Articular / Q000166:cytology*; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics
D016207:Cytokines; D007375:Interleukin-1; D012313:RNA