DEVELOPMENT OF A REAL-TIME RT-PCR-BASED TEST PANEL FOR ASSESSMENT OF CYTOKINE PROFILE IN MONONUCLEAR CELLS FROM BLOOD AND SYNOVIAL FLUID IN RHEUMATOID ARTHRITIS

Abstract. An imbalance between pro- and anti-inflammatory cytokine synthesis plays a crucial role in pathogenesis of rheumatoid arthritis (RA), including erosive joint lesions. Differences in cytokine profiles of RA patients are sufficiently investigated. However, possible interrelations between cytokine profile, immune inflammation, RA progression, and the disease prognosis require further investigations. Due to active search for novel targets in anti-cytokine therapy of RA, evaluation of cytokine levels in peripheral blood and synovial joint fluid remain quite relevant. Therefore, novel methods aimed to determine mRNA expressed by cytokine genes are thought to be promising. Our research was intended to develop test systems for quantitative determination of mRNAs for the following cytokines: TNFα, IL-1β, IL-6, IL-8, IL-17, IL-15, IL-10, IFNγ, IL-4, IL-2. Present article concerns specificities of the developed test systems, description of their technical principles, as well as comparative studies of cytokine gene expression in peripheral blood and synovial joint fluid in RA patients. (Med. Immunol., 2008, vol. 10, N 6, pp 563-570).

1. Насонов E.Л., Гукасян Д.А., Насонова М.Б. Иммунопатология ревматоидного артрита и остеопороз: новые данные // Остеопороз и остеопатии. – 2000. – № 2. – С. 4-7.

2. Abbas A.K. Murphy K.M., Sher A. Functional diversity of T helper lymphocytes // Nature. – 1996. – Vol. 383. – P. 787-793.

3. Arnett F.C., Edworthy S.M., Bloch D.A., McShane D.J., Fries J.F., Cooper N.S., Healy L.A., Kaplan S.R., Liang M.H., Luthra H.S. at al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis // Arthritis Rheum. – 1988. – Vol. 31. – P.315-324.

4. Dolhain R.J., van der Heiden A.N., ter Haar N.T. Breedveld F.C., Miltenburg A.M. Shift toward T lymphocytes with a T helper 1 cytokine-secretion profile in the joints of patient with rheumatoid arthritis // Arthritis Rheum. – 1996. – Vol. 39. – P. 1961-1969.

5. Gracie J.A., Forsey R.J., Chan W.L., Gilmour A., Leung B.P., Greer M.R., Kennedy K., Carter R., Wei X.Q., Xu D., Field M., Foulis A., Liew F.Y., McInnes I.B. A proinflammatory role for IL-18 in rheumatoid arthritis // J Clin Invest. – 1999. – Vol. 104. – P. 1393 - 1401.

6. Gaffen S.L. Biology of recently discovered cytokines: Interleukin-17 – a unique inflammatory cytokine with roles in bone biology and arthritis // Arthritis Res Ther. – 2004. – Vol. 6. – P. 240-247.

7. Kim K-W., Cho M-L, Park M-K., Yoon C.H., Park S.H., Lee S.H., Kim H.Y. Increased interleukin-17 production via a phosphoinositide 3-kinase/Akt and nuclear factor kB-dependent pathway in patients with rheumatoid arthritis // Arthritis Res Ther. – 2005. – Vol. 7.– P. 139-148.

8. McInnes I.B., Gracie J.A., Harnett M., Harnett W., Liew F.Y. New strategies to control inflammatory synovitis: interleukin 15 and beyond // Ann Rheum Dis. – 2003. – Vol. 62. – P. 51-54.

9. McInnes I.B., Gracie J.A. Interleukin 15: a proinflammatory role in rheumatoid arthritis synovitis // Immunol Today. – 1998. – Vol. 19. – P. 75-79.

10. Miosses P.,van den Berg W. Th1/Th2 cytokine balance inarthritis // Arthritis Rheum. – 1997. – Vol. 40. – P. 2105-2115.

11. Morita Y., Yamamura M., Kawashima M. Harada S., Tsuji K., Shibuya K., Maruyama K., Makino H. Flow cytometric singl-cell analisis of cytokine production by CD4+ T cell in synovial tissue and periferal blood from patients with rheumatoid arthritis // Arthritis Rheum. – 1998. – Vol. 41. – P. 1669 - 1676.

12. Park J.Y., Pillinger M.H. Interleukin-6 the pathogenesis of rheumatoid arthritis // Bull NYU Hosp Joint Dis. – 2007. – Vol. 65. – P. 4-10.

13. Simon A.K., Seipelt E., Sieper J. Divergent T-cell cytokine patterns in iflammatory arthritis // Proc Natl Acad Sci USA. – 1994. – Vol. 91. – P. 8562 - 8566.

14. Van Roon J.A., van Roy J.L.,Duits A., Lafeber F.P., Bijlsma J.W. Proinflammatory cytokine production and cartilage damage due to rheumatoid synovial T-helper-1 activition is inhibited by interleukin-4 // Ann Rheum Dis. – 1995. – Vol.54. – P. 836-840.