Cytokine mRNA repertoire of articular chondrocytes from arthritic patients, infants, and neonatal mice
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Articular chondrocytes from nine arthritic patients, five infants, and Balb/c neonatal mice were analyzed for the presence of various cytokine mRNAs by a reverse transcriptase polymerase chain reaction (RT-PCR). Four cytokine mRNAs, interleukin (IL)-6, IL-8, IL-11, and macrophage colony stimulating factor (M-CSF), were detected in all human chondrocytes, regardless of source. IL-10, IL-12p35, and tumor necrosis factor alpha (TNF-α) transcripts were found in at least 12 of the 14 human samples. IL-13, granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating factor (GMCSF), and TNF-βmRNAs were found more predominantly in infant samples and in samples from patients with rheumatoid arthritis (RA) compared with samples from patients with osteoarthritis (OA). Another group of cytokine mRNAs, IL-1 (α, β), IL-4, IL-5, and IL-7, were only weakly expressed in some human samples. The cytokine transcripts that were not found were IL-2, IL-3, and interferon gamma (IFN-γ. Because of the large array of cytokine transcripts detected, human chondrocyte preparations were further purified by reacting them with a monoclonal antibody specific to chondrocyte differentiation antigen and subjecting them to fluorescent-activated cell sorting. A similar array of cytokines was found between the sorted and unsorted chondrocytes, although TNF-α, G-CSF, and GM-CSF transcripts appeared to be upregulated during the sorting process. Human chondrocytes that dedifferentiated into fibroblasts (a 40-day and a 77-day culture) no longer expressed mRNAs for IL-1, G-CSF, GM-CSF, and TNF-α, but all other cytokine mRNAs remained detectable. Although certain phenotypic characteristics were lost:, including reactivity to chondrocyte-specific monoclonal antibodies and morphological features, chondrocytes in long-term culture still expressed cytokine mRNAs. As expected, more consistent results were obtained when seven preparations of chondrocytes from neonatal Balb/c mice were examined using available cytokine primers. They contained IL-1, IL-5, IL-6, IL-7, IL-12, GM-CSF, M-CSF, transforming growth factor beta (TGF-β), TNF-α, and TNF-β mRNAs but lacked IL-2, IL-3, IL-4, IL-10, and IFN-γ mRNAs. Future experiments to define conditions by which these cytokine protein products are expressed are needed to help assess their roles in chondrocyte biology and in disease states.
Key wordsCytokine mRNAs Articular chondrocytes Rheumatoid arthritis Osteoarthritis Infants Neonatal mice
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- 10.Alsalameh S, Firestein GS, Oez S, Kurrle R, Kalden JR, Burmester GR (1994) Regulation of granulocyte macrophage colony stimulating factor production by human articular chondrocytes. Induction by both tumor necrosis factor-α and interleukin 1, downregulation by transforming growth factorβ and upregulation by fibroblast growth factor. J Rheumatol 21:993–1002PubMedGoogle Scholar
- 13.Chacko S, Abbott J, Holtzer S, Holtzer H (1969) The loss of phenotypic traits by differentiated cell. J Exp Med 134:417–442Google Scholar
- 15.Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries IF, Cooper NS, Healey LA, Kaplan SR, Liang MH, Luthr HS, Medsger TA Jr, Mitchell DM, Neustadt DH, Pinals RS, Schaller JG, Sharp IT, Wilder RL, Hunder GG (1988) The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 31:315–324PubMedGoogle Scholar
- 17.Yokoyama WM (1991) Cryopreservation of cells. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W (eds) Current protocols in immunology. Wiley, New York, pp A3.15-A3.17Google Scholar
- 18.Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Plainview, NY, pp 6.24–6.27Google Scholar
- 20.Tanabe B (1994) Cytokine repertoire of human articular chondrocytes in relationship to culture conditions and dedifferentiation. MS thesis, University of HawaiiGoogle Scholar
- 25.Thomson AW (1994) The cytokine handbook, 2nd edn. Academic Press, San Diego, Calif.Google Scholar
- 27.Trichieri G (1975) Interleukin 12: a proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. Ann Rev Immunol 13:251–256Google Scholar
- 28.Minty A, Chalon P, Derocq JM, Dumont X, Guillemont JC, Kahad M, Labit C, Leplatois P, Liauzun P, Miloux B, Minty C, Casellas P, Laison G, Lupber J, Dhire D, Ferrara P, Caput D (1993) Interleukin 13 is a new human lymphokine regulating inflammatory and immune responses. Nature 362:248–250PubMedGoogle Scholar