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  • 제13권1호
  • /
  • Pages.16-24
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  • 2013
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  • 1598-2629(pISSN)
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  • 2092-6685(eISSN)
대한면역학회 (The Korean Association of Immunobiologists)
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DOI QR코드

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Functional Characteristics of C-terminal Lysine to Cysteine Mutant Form of CTLA-4Ig

  • Kim, Bongi (Department of Microbiology and Immunology, Seoul National University College of Medicine) ;
  • Shin, Jun-Seop (Department of Microbiology and Immunology, Seoul National University College of Medicine) ;
  • Park, Chung-Gyu (Department of Microbiology and Immunology, Seoul National University College of Medicine)
  • 투고 : 2013.01.21
  • 심사 : 2013.02.12
  • 발행 : 2013.02.28
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초록

CTLA-4Ig is regarded as an inhibitory agent of the T cell proliferation via blocking the costimulatory signal which is essential for full T cell activation. To improve applicability, we developed the CTLA-4Ig-CTKC in which the c-terminal lysine had been replaced by cysteine through single amino acid change. The single amino acid mutation of c-terminus of CTLA-4Ig was performed by PCR and was checked by in vitro transcription and translation. DNA construct of mutant form was transfected to Chinese hamster ovary (CHO) cells by electroporation. The purified proteins were confirmed by Western blot and B7-1 binding assay for their binding ability. The suppressive capacity of CTLA-4Ig-CTKC was evaluated by the mixed lymphocyte reaction (MLR) and in the allogeneic pancreatic islet transplantation model. CTLA-4Ig-CTKC maintained binding ability to B7-1 molecule and effectively inhibits T cell proliferation in MLR. In the murine allogeneic pancreatic islet transplantation, short-term treatment of CTLA-4Ig-CTKC prolonged the graft survival over 100 days. CTLA-4Ig-CTKC effectively inhibits immune response both in MLR and in allogeneic islet transplantation model, indicating that single amino acid mutation does not affect the inhibitory function of CTLA-4Ig. CTLA-4Ig-CTKC can be used in vehicle-mediated drug delivery system such as liposome conjugation.

키워드

참고문헌

  1. Lenschow, D. J., T. L. Walunas, and J. A. Bluestone. 1996. CD28/B7 system of T cell costimulation. Annu. Rev. Immunol. 14: 233-258. https://doi.org/10.1146/annurev.immunol.14.1.233
  2. Lafferty, K. J., S. J. Prowse, C. J. Simeonovic, and H. S. Warren. 1983. Immunobiology of tissue transplantation: a return to the passenger leukocyte concept. Annu. Rev. Immunol. 1: 143-173. https://doi.org/10.1146/annurev.iy.01.040183.001043
  3. June, C. H., J. A. Bluestone, L. M. Nadler, and C. B. Thompson. 1994. The B7 and CD28 receptor families. Immunol. Today 15: 321-331. https://doi.org/10.1016/0167-5699(94)90080-9
  4. Salomon, B. and J. A. Bluestone. 2001. Complexities of CD28/B7: CTLA-4 costimulatory pathways in autoimmunity and transplantation. Annu. Rev. Immunol. 19: 225-252. https://doi.org/10.1146/annurev.immunol.19.1.225
  5. Greenfield, E. A., K. A. Nguyen, and V. K. Kuchroo. 1998. CD28/B7 costimulation: a review. Crit. Rev. Immunol. 18: 389-418. https://doi.org/10.1615/CritRevImmunol.v18.i5.10
  6. Walunas, T. L., D. J. Lenschow, C. Y. Bakker, P. S. Linsley, G. J. Freeman, J. M. Green, C. B. Thompson, and J. A. Bluestone. 1994. CTLA-4 can function as a negative regulator of T cell activation. Immunity 1: 405-413. https://doi.org/10.1016/1074-7613(94)90071-X
  7. Teft, W. A., M. G. Kirchhof, and J. Madrenas. 2006. A molecular perspective of CTLA-4 function. Annu. Rev. Immunol. 24: 65-97. https://doi.org/10.1146/annurev.immunol.24.021605.090535
  8. Lenschow, D. J., Y. Zeng, J. R. Thistlethwaite, A. Montag, W. Brady, M. G. Gibson, P. S. Linsley, and J. A. Bluestone. 1992. Long-term survival of xenogeneic pancreatic islet grafts induced by CTLA4lg. Science 257: 789-792. https://doi.org/10.1126/science.1323143
  9. Turka, L. A., P. S. Linsley, H. Lin, W. Brady, J. M. Leiden, R. Q. Wei, M. L. Gibson, X. G. Zheng, S. Myrdal, and D. Gordon, et al. 1992. T-cell activation by the CD28 ligand B7 is required for cardiac allograft rejection in vivo. Proc. Natl. Acad. Sci. U. S. A. 89: 11102-11105. https://doi.org/10.1073/pnas.89.22.11102
  10. Li, W., L. Lu, Z. Wang, L. Wang, J. J. Fung, A. W. Thomson, and S. Qian. 2001. Costimulation blockade promotes the apoptotic death of graft-infiltrating T cells and prolongs survival of hepatic allografts from FLT3L-treated donors. Transplantation 72: 1423-1432. https://doi.org/10.1097/00007890-200110270-00016
  11. Larsen, C. P., T. C. Pearson, A. B. Adams, P. Tso, N. Shirasugi, E. Strobert, D. Anderson, S. Cowan, K. Price, J. Naemura, J. Emswiler, J. Greene, L. A. Turk, J. Bajorath, R. Townsend, D. Hagerty, P. S. Linsley, and R. J. Peach. 2005. Rational development of LEA29Y (belatacept), a high-affinity variant of CTLA4-Ig with potent immunosuppressive properties. Am. J. Transplant. 5: 443-453. https://doi.org/10.1111/j.1600-6143.2005.00749.x
  12. Park, C. G., N. W. Thiex, K. M. Lee, G. L. Szot, J. A. Bluestone, and K. D. Lee. 2003. Targeting and blocking B7 costimulatory molecules on antigen-presenting cells using CTLA4Ig-conjugated liposomes: in vitro characterization and in vivo factors affecting biodistribution. Pharm. Res. 20: 1239- 1248. https://doi.org/10.1023/A:1025057216492
  13. Lacy, P. E. and M. Kostianovsky. 1967. Method for the isolation of intact islets of Langerhans from the rat pancreas. Diabetes 16: 35-39.
  14. Abrams, J. R., M. G. Lebwohl, C. A. Guzzo, B. V. Jegasothy, M. T. Goldfarb, B. S. Goffe, A. Menter, N. J. Lowe, G. Krueger, M. J. Brown, R. S. Weiner, M. J. Birkhofer, G. L. Warner, K. K. Berry, P. S. Linsley, J. G. Krueger, H. D. Ochs, S. L. Kelley, and S. Kang. 1999. CTLA4Ig-mediated blockade of T-cell costimulation in patients with psoriasis vulgaris. J. Clin. Invest. 103: 1243-1252. https://doi.org/10.1172/JCI5857
  15. Genovese, M. C., J. C. Becker, M. Schiff, M. Luggen, Y. Sherrer, J. Kremer, C. Birbara, J. Box, K. Natarajan, I. Nuamah, T. Li, R. Aranda, D. T. Hagerty, and M. Dougados. 2005. Abatacept for rheumatoid arthritis refractory to tumor necrosis factor alpha inhibition. N. Engl. J. Med. 353: 1114- 1123. https://doi.org/10.1056/NEJMoa050524
  16. Kremer, J. M., R. Westhovens, M. Leon, E. Di Giorgio, R. Alten, S. Steinfeld, A. Russell, M. Dougados, P. Emery, I. F. Nuamah, G. R. Williams, J. C. Becker, D. T. Hagerty, and L. W. Moreland. 2003. Treatment of rheumatoid arthritis by selective inhibition of T-cell activation with fusion protein CTLA4Ig. N. Engl. J. Med. 349: 1907-1915. https://doi.org/10.1056/NEJMoa035075
  17. Kaplan, B. 2010. Belatacept: the promises and challenges of belatacept and costimulatory blockade. Am. J. Transplant. 10: 441-442. https://doi.org/10.1111/j.1600-6143.2010.03026.x
  18. Larsen, C. P., J. Grinyó, J. Medina-Pestana, Y. Vanrenterghem, F. Vincenti, B. Breshahan, J. M. Campistol, S. Florman, C. Rial Mdel, N. Kamar, A. Block, G. Di Russo, C. S. Lin, P. Garg, and B. Charpentier. 2010. Belatacept-based regimens versus a cyclosporine A-based regimen in kidney transplant recipients: 2-year results from the BENEFIT and BENEFIT-EXT studies. Transplantation 90: 1528-1535. https://doi.org/10.1097/TP.0b013e3181ff87cd
  19. Vincenti, F., G. Blancho, A. Durrbach, P. Friend, J. Grinyo, P. F. Halloran, J. Klempnauer, P. Lang, C. P. Larsen, F. Mühlbacher, B. Nashan, J. P. Soulillou, Y. Vanrenterghem, T. Wekerle, M. Agarwal, S. Gujrathi, J. Shen, R. Shi, R. Townsend, and B. Charpentier. 2010. Five-year safety and efficacy of belatacept in renal transplantation. J. Am. Soc. Nephrol. 21: 1587-1596. https://doi.org/10.1681/ASN.2009111109
  20. Vincenti, F., B. Charpentier, Y. Vanrenterghem, L. Rostaing, B. Bresnahan, P. Darji, P. Massari, G. A. Mondragon-Ramirez, M. Agarwal, G. Di Russo, C. S. Lin, P. Garg, and C. P. Larsen. 2010. A phase III study of belatacept-based immunosuppression regimens versus cyclosporine in renal transplant recipients (BENEFIT study). Am. J. Transplant. 10: 535-546. https://doi.org/10.1111/j.1600-6143.2009.03005.x
  21. Durrbach, A., J. M. Pestana, T. Pearson, F. Vincenti, V. D. Garcia, J. Campistol, C. Rial Mdel, S. Florman, A. Block, G. Di Russo, J. Xing, P. Garg, and J. Grinyó. 2010. A phase III study of belatacept versus cyclosporine in kidney transplants from extended criteria donors (BENEFIT-EXT study). Am. J. Transplant. 10: 547-557. https://doi.org/10.1111/j.1600-6143.2010.03016.x
  22. Alegre, M. L., J. Y. Tso, H. A. Sattar, J. Smith, F. Desalle, M. Cole, and J. A. Bluestone. 1995. An anti-murine CD3 monoclonal antibody with a low affinity for Fc gamma receptors suppresses transplantation responses while minimizing acute toxicity and immunogenicity. J. Immunol. 155: 1544-1555.

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