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The Korean Association of Immunobiologists (대한면역학회)
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Highly Efficient Gene Expression in Rabbit Synoviocytes Using EBV-Based Plasmid

가토 윤활막 세포에서 EBV-Based 플라스미드를 사용한 효율적인 유전자 발현

  • Kim, Jin Young (Research Institute of Immunobiology, Catholic Research Institutes of Medical Science, Catholic University of Korea) ;
  • Oh, Sang Taek (Research Institute of Immunobiology, Catholic Research Institutes of Medical Science, Catholic University of Korea) ;
  • Youn, JeeHee (Department of Anatomy and Cell Biology, Hanyang University School of Medicine) ;
  • Lee, Suk Kyeong (Research Institute of Immunobiology, Catholic Research Institutes of Medical Science, Catholic University of Korea)
  • 김진영 (가톨릭대학교 의과학연구원 면역생물학연구소) ;
  • 오상택 (가톨릭대학교 의과학연구원 면역생물학연구소) ;
  • 윤지희 (한양대학교 의과대학 해부학교실) ;
  • 이숙경 (가톨릭대학교 의과학연구원 면역생물학연구소)
  • Published : 2004.09.30
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Abstract

Background: Rheumatoid arthritis (RA) is an autoimmune disorder characterized by chronic synovial inflammation which leads to joint destruction. Gene therapy of RA targets the players of inflammation or articular destruction. However, viral vectors have safety problems and side effects, while non-viral vectors suffer from inefficient gene transfer and fast loss of gene expression. To overcome the limits of non-vial vectors, an EBV-based plasmid which is known to exert prolonged high level gene expression can be used. Methods: pEBVGFP, pEBVIL-10, and pEBVvIL-10 were constructed by cloning GFP, IL-10, and vIL-10 genes into an EBV-based plasmid, respectively. The pGFP was used as a control plasmid. Each constructs were lipofected into HIG-82 rabbit synoviocytes. The expression of GFP was monitored by FACS and confocal microscopy. IL-10 and vIL-10 expressions were measured by ELISA. Results: GFP expression 2 days after transfection was achieved in 33.2% of cells. GFP-expressing cells transfected with pGFP decreased rapidly from 4 days after transfection and disappeared completely by 11 days. Cells transfected with pEBVGFP began to decrease slowly from 4 days. But GFP expression was detected for over 35 days. In addition, HIG-82 cells transfected with pEBVIL-10 ($44.6{\pm}1.5ng/ml$) or pEBVvIL-10 ($51.0{\pm}5.7ng/ml$) secreted these cytokines at high levels. High level cytokine production by hygromycin selection was maintained at least for up to 26 days after transfection. Conclusion: These results suggest that the EBV-based plasmid has a potential to improve non-viral gene transfer system and may be applicable to treat RA without the drawbacks of viral vectors.

Keywords

References

  1. Feldmann M, Brennan FM, Maini RN: Rheumatoid Arthritis. Cell 85;307-310, 1996 https://doi.org/10.1016/S0092-8674(00)81109-5
  2. Robbins PD, Evans CH, Chernajovsky Y: Gene therapy for arthritis. Gene Ther 10;902-911, 2003 https://doi.org/10.1038/sj.gt.3302040
  3. Feldmann M, Brennan FM, Maini, RN: Role of cytokines in rheumatoid arthritis. Annu Rev Immunol 14;397-440, 1996 https://doi.org/10.1146/annurev.immunol.14.1.397
  4. Vervoordeldonk MJ, Tak PP: Gene therapy in rheumatic diseases. Best Pract Res Clin Rheumatol 15;771-788, 2001 https://doi.org/10.1053/berh.2001.0193
  5. Afeltra A: Treatment of Rheumatoid arthritis: New Therapeutic Approaches with Biological Agents. Curr Drug Targets Immune Endocr Metabol Disord 1;45-65, 2001 https://doi.org/10.2174/1568008013341677
  6. Boissier MC, Bessis N: Therapeutic gene transfer for rheumatoid arthritis. Reumatismo 56;51-61, 2004
  7. Bessis N, Doucet C, Cottard V, Douar AM, Firat H, Jorgensen C, Mezzina M, Boissier MC: Gene therapy for rheumatoid arthritis. J Gene Med 4;581-591, 2002 https://doi.org/10.1002/jgm.325
  8. Ghivizzani SC, Oligino TJ, Glorioso JC, Robbins PD, Evans CH: Direct gene delivery strategies for the treatment of rheumatoid arthritis. Drug Discov Today 6;259-267, 2001 https://doi.org/10.1016/S1359-6446(01)01685-3
  9. Middleton T, Sugden B: Retention of plasmid DNA in mammalian cells is enhanced by binding of the Epstein-Barr virus replication protein EBNA1. J Virol 68;4067-4071, 1994
  10. Mazda O: Improvement of Nonviral Gene Therapy by Epstein-Barr Virus (EBV)-based Plasmid Vectors. Curr Gene Ther 2;379-392, 2002 https://doi.org/10.2174/1566523023347814
  11. Mucke S, Polack A, Pawlita M, Zehnpfennig D, Massoudi N, Bohlen H, Doerfler W, Bornkamm G, Diehl V, Wolf J: Suitability of Epstein-Barr virus-based episomal vectors for expression of cytokine genes in human lymphoma cells. Gene Ther 4;82-92, 1997 https://doi.org/10.1038/sj.gt.3300363
  12. Min KA, Oh ST, Yoon KH, Kim CK, Lee SK: Prolonged gene expression in primary porcine pancreatic cells using an Epstein-Barr virus-based episomal vector. BBRC 305;108-115, 2003
  13. Zhang J, Wilson A, Alber S, Ma Z, Tang ZL, Satoh E, Mazda O, Watkins S, Huang L, Pitt B, Li S: Prolonged gene expression in mouse lung endothelial cells following transfection with Epstein-Barr virus-based episomal plasmid. Gene Ther 10;822-826, 2003 https://doi.org/10.1038/sj.gt.3301958
  14. Taga K, Tosato G: IL-10 inhibits human T cell proliferation and IL-2 production. J Immunol 148;1143-1148, 1992
  15. Schulze-Koops H, Kalden JR: The balance of Th1/Th2 cytokines in rheumatoid arthritis. Best Pract Res Clin Rheumatol 15;677-691, 2001 https://doi.org/10.1053/berh.2001.0187
  16. Katsikis PD, Chu CQ, Brennan FM, Maini RN, Feldmann M: Immunoregulatory role of interleukin 10 in rheumatoid arthritis. J Exp Med 179;1517-1527, 1994 https://doi.org/10.1084/jem.179.5.1517
  17. Ma Y, Thornton S, Duwel LE, Boivin GP, Giannini EH, Leiden JM, Bluestone JA, Hirsch R: Inhibition of Collagen- Induced Arthritis in Mice by Viral IL-10 Gene Transfer. J Immunol 161;1516-1524, 1998
  18. Evans CH, Ghivizzani SC, Kang R, Muzzonigro T, Wasko MC, Herndon JH, Robbins PD: Gene therapy for rheumatic diseases. Arthritis Rheum 42;1-16, 1999 https://doi.org/10.1002/1529-0131(199901)42:1<1::AID-ANR1>3.0.CO;2-4
  19. Langle-Rouault F, Patzel V, Benavente A, Taillez M, Silvestre N, Bompard A, Sczakiel G, Jacobs E, Rittner K: Up to 100-Fold Increase of Apparent Gene Expression in the Presence of Epstein-Barr Virus oriP Sequences and EBNA1: Implications of the Nuclear Import of Plasmids. J Virol 72;6181-6185, 1998
  20. Otani K, Nita I, Macaulay W, Georgescu HI, Robbins PD, Evans CH: Suppression of antigen-induced arthritis in rabbits by ex vivo gene therapy. J Immunol 156;3558-3562, 1996
  21. Henderson B, Glynn LE: Metabolic alterations in the synoviocytes in chronically inflamed knee joints in immune arthritis in the rabbits: comparison with rheumatoid arthritis. Br J Exp Pathol. 62;27-33, 1981
  22. 오상택, 민경아, 김종국, 이숙경: HSV-TK 유전자를 암호화하는 EBV 유래 플라스미드를 이용한 유전자 치료. 약제학회지 33;267-272, 2003
  23. Lubberts E, Joosten LA, Van Den Bersselaar L, Helsen MM, Bakker AC, Xing Z, Richards CD, Van Den Berg WB: Intraarticular IL-10 gene transfer regulates the expression of collagen-induced arthritis (CIA) in the knee and ipsilateral paw. Clin Exp Immunol 120;375-383, 2000 https://doi.org/10.1046/j.1365-2249.2000.01217.x
  24. Enzmann V, Hollborn M, Poschinger K, Wiedemann P, Kohen L: Immunosupperss-ion by IL-10-transfected human retinal pigment epithelialcells in vitro. Curr Eye Res 23;98-105, 2001 https://doi.org/10.1076/ceyr.23.2.98.5474
  25. Sasaki M, Jordan P, Houghton J, Meng X, Itoh M, Joh T, Alexander JS: Transfection of IL-10 expression vectors into endothelial cultures attenuates alpha4beta7-dependent lymphocyte adhesion mediated by MAdCAM-1. BMC Gastroenterol 3;3-10, 2003 https://doi.org/10.1186/1471-230X-3-3