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Construction, and In Vitro and In Vivo Analyses of Tetravalent Immunoadhesins

  • Cho, Hoonsik (Department of Microbiology, College of Medicine, Hanyang University) ;
  • Chung, Yong-Hoon (Department of Microbiology, College of Medicine, Hanyang University)
  • Received : 2012.01.17
  • Accepted : 2012.04.14
  • Published : 2012.08.28

Abstract

Previous observations demonstrated that various immunosuppressive agents and their combination therapies can increase allograft survival rates. However, these treatments may have serious side effects and cannot substantially improve or prolong graft survival in acute graft-versus-host disease (GVHD). To improve the therapeutic potency of divalent immunoadhesins, we have constructed and produced several tetravalent forms of immunoadhesins comprising each of cytotoxic T-lymphocyte-associated antigen-4 (CTLA4), CD2, and lymphocyte activation gene-3 (LAG3). Flow cytometric and T cell proliferation analyses displayed that tetravalent immunoadhesins have a higher binding affinity and more potent efficacy than divalent immunoadhesins. Although all tetravalent immunoadhesins possess better efficacies, tetravalent forms of CTLA4-Ig and LAG3-Ig revealed higher inhibitory effects on T cell proliferation than tetravalent forms of TNFR2-Ig and CD2-Ig. In vitro mixed lymphocytes reaction (MLR) showed that combined treatment with tetravalent CTLA4-Ig and tetravalent LAG3-Ig was highly effective for inhibiting T cell proliferation in both human and murine allogeneic stimulation. In addition, both single tetravalent-form and combination treatments can prevent the lethality of murine acute GVHD. The results of this study demonstrated that co-blockade of the major histocompatibility complex class (MHC)II:T cell receptor (TCR) and CD28:B7 pathways by using tetravalent human LAG3-Ig and CTLA4-Ig synergistically prevented murine acute GVHD.

Acknowledgement

Supported by : Ministry of Health & Welfare

References

  1. Adams, A. B., N. Shirasugi, M. M. Durham, E. Strobert, D. Anderson, P. Rees, et al. 2002. Calcineurin inhibitor-free CD28 blockade-based protocol protects allogeneic islets in nonhuman primates. Diabetes 51: 265-270. https://doi.org/10.2337/diabetes.51.2007.S265
  2. Adams, G. P., C. C. Shaller, L. L. Chappell, C. Wu, E. M. Horak, H. H. Simmons, et al. 2000. Delivery of the alphaemitting radioisotope bismuth-213 to solid tumors via singlechain Fv and diabody molecules. Nucl. Med. Biol. 27: 339-346. https://doi.org/10.1016/S0969-8051(00)00103-7
  3. Antin, J. H. and J. L. Ferrara. 1992. Cytokine dysregulation and acute graft-versus-host disease. Blood 80: 2964-2968.
  4. Benito, A. I., T. Furlong, P. J. Martin, C. Anasetti, F. R. Appelbaum, K. Doney, et al. 2001. Sirolimus (rapamycin) for the treatment of steroid-refractory acute graft-versus-host disease. Transplantation 72: 1924-1929. https://doi.org/10.1097/00007890-200112270-00010
  5. Bierer, B. E., G. Hollander, D. Fruman, and S. J. Burakoff. 1993. Cyclosporin A and FK506: Molecular mechanisms of immunosuppression and probes for transplantation biology. Curr. Opin. Immunol. 5: 763-773. https://doi.org/10.1016/0952-7915(93)90135-F
  6. Blazar, B. R., P. A. Taylor, P. S. Linsley, and D. A. Vallera. 1994. In vivo blockade of CD28/CTLA4: B7/BB1 interaction with CTLA4-Ig reduces lethal murine graft-versus-host disease across the major histocompatibility complex barrier in mice. Blood 83: 3815-3825.
  7. Blazar, B. R., P. A. Taylor, A. Panoskaltsis-Mortari, G. S. Gray, and D. A. Vallera. 1995. Coblockade of the LFA1:ICAM and CD28/CTLA4:B7 pathways is a highly effective means of preventing acute lethal graft-versus-host disease induced by fully major histocompatibility complex-disparate donor grafts. Blood 85: 2607-2618.
  8. Bluestone, J. A., E. W. St Clair, and L. A. Turka. 2006. CTLA4Ig: Bridging the basic immunology with clinical application. Immunity 24: 233-238. https://doi.org/10.1016/j.immuni.2006.03.001
  9. Capon, D. J., S. M. Chamow, J. Mordenti, S. A. Marsters, T. Gregory, H. Mitsuya, et al. 1989. Designing CD4 immunoadhesins for AIDS therapy. Nature 337: 525-531. https://doi.org/10.1038/337525a0
  10. Comoli, P., F. Locatelli, A. Moretta, D. Montagna, V. Calcaterra, A. Cometa, et al. 2001. Human alloantigen-specific anergic cells induced by a combination of CTLA4-Ig and CsA maintain anti-leukemia and anti-viral cytotoxic responses. Bone Marrow Transplant. 27: 1263-1273. https://doi.org/10.1038/sj.bmt.1703063
  11. Economides, A. N., L. R. Carpenter, J. S. Rudge, V. Wong, E. M. Koehler-Stec, C. Hartnett, et al. 2003. Cytokine traps: Multicomponent, high-affinity blockers of cytokine action. Nat. Med. 9: 47-52.
  12. Emery, P. 2003. The therapeutic potential of costimulatory blockade with CTLA4Ig in rheumatoid arthritis. Expert Opin. Investig. Drugs 12: 673-681. https://doi.org/10.1517/13543784.12.4.673
  13. Goldenberg, M. M. 1999. Etanercept, a novel drug for the treatment of patients with severe, active rheumatoid arthritis. Clin. Ther. 21: 75-87; discussion 1-2. https://doi.org/10.1016/S0149-2918(00)88269-7
  14. Gottlieb, A. B. 2004. Etanercept for the treatment of psoriasis and psoriatic arthritis. Dermatol. Ther. 17: 401-408. https://doi.org/10.1111/j.1396-0296.2004.04043.x
  15. Grakoui, A., S. K. Bromley, C. Sumen, M. M. Davis, A. S. Shaw, P. M. Allen, and M. L. Dustin. 1999. The immunological synapse: A molecular machine controlling T cell activation. Science 285: 221-227. https://doi.org/10.1126/science.285.5425.221
  16. Gratama, J. W., J. Jansen, R. A. Lipovich, H. J. Tanke, G. Goldstein, and F. E. Zwaan. 1984. Treatment of acute graftversus-host disease with monoclonal antibody OKT3. Clinical results and effect on circulating T lymphocytes. Transplantation 38: 469-474. https://doi.org/10.1097/00007890-198411000-00005
  17. Guidotti, L. G. and F. V. Chisari. 2000. Cytokine-mediated control of viral infections. Virology 273: 221-227. https://doi.org/10.1006/viro.2000.0442
  18. Guinan, E. C., J. G. Gribben, V. A. Boussiotis, G. J. Freeman, and L. M. Nadler. 1994. Pivotal role of the B7:CD28 pathway in transplantation tolerance and tumor immunity. Blood 84: 3261-3282.
  19. Guy-Grand, D., M. Dy, G. Luffau, and P. Vassalli. 1986. Gut mucosal mast cells: Origin, traffic and differentiation in mice and rats. Ann. Inst. Pasteur Immunol. 137D: 215-222.
  20. Hale, D. A., R. Gottschalk, T. Maki, and A. P. Monaco. 1997. Use of CTLA4-Ig in combination with conventional immunosuppressive agents to prolong allograft survival. Transplantation 64: 897-900. https://doi.org/10.1097/00007890-199709270-00018
  21. Hanahan, D. 1983. Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166: 557-580. https://doi.org/10.1016/S0022-2836(83)80284-8
  22. Harlan, D. M., R. Abe, K. P. Lee, and C. H. June. 1995. Potential roles of the B7 and CD28 receptor families in autoimmunity and immune evasion. Clin. Immunol. Immunopathol. 75: 99-111. https://doi.org/10.1006/clin.1995.1058
  23. Harty, J. T., A. R. Tvinnereim, and D. W. White. 2000. CD8+ T cell effector mechanisms in resistance to infection. Annu. Rev. Immunol. 18: 275-308. https://doi.org/10.1146/annurev.immunol.18.1.275
  24. Huard, B., P. Prigent, F. Pages, D. Bruniquel, and F. Triebel. 1996. T cell major histocompatibility complex class II molecules down-regulate CD4+ T cell clone responses following LAG-3 binding. Eur. J. Immunol. 26: 1180-1186. https://doi.org/10.1002/eji.1830260533
  25. Huard, B., M. Tournier, T. Hercend, F. Triebel, and F. Faure. 1994. Lymphocyte-activation gene 3/major histocompatibility complex class II interaction modulates the antigenic response of CD4+ T lymphocytes. Eur. J. Immunol. 24: 3216-3221. https://doi.org/10.1002/eji.1830241246
  26. Jacobsohn, D. A. and G. B. Vogelsang. 2002. Novel pharmacotherapeutic approaches to prevention and treatment of GVHD. Drugs 62: 879-889. https://doi.org/10.2165/00003495-200262060-00002
  27. Kaplon, R. J., P. S. Hochman, R. E. Michler, P. A. Kwiatkowski, N. M. Edwards, C. L. Berger, et al. 1996. Short course single agent therapy with an LFA-3-IgG1 fusion protein prolongs primate cardiac allograft survival. Transplantation 61: 356-363. https://doi.org/10.1097/00007890-199602150-00004
  28. Kirk, A. D., D. M. Harlan, N. N. Armstrong, T. A. Davis, Y. Dong, G. S. Gray, et al. 1997. CTLA4-Ig and anti-CD40 ligand prevent renal allograft rejection in primates. Proc. Natl. Acad. Sci. USA 94: 8789-8794. https://doi.org/10.1073/pnas.94.16.8789
  29. Kortt, A. A., O. Dolezal, B. E. Power, and P. J. Hudson. 2001. Dimeric and trimeric antibodies: High avidity scFvs for cancer targeting. Biomol. Eng. 18: 95-108. https://doi.org/10.1016/S1389-0344(01)00090-9
  30. Krieger, N. R., D. Yuh, W. B. McIntyre, T. F. Flavin, D. Yin, R. Robbins, and C. G. Fathman. 1998. Prolongation of cardiac graft survival with anti-CD4Ig plus hCTLA4Ig in primates. J. Surg. Res. 76: 174-178. https://doi.org/10.1006/jsre.1998.5318
  31. Lafferty, K. J. 1994. A contemporary view of transplantation tolerance: An immunologist's perspective. Clin. Transplant. 8: 181-187.
  32. Larsen, C. P., E. T. Elwood, D. Z. Alexander, S. C. Ritchie, R. Hendrix, C. Tucker-Burden, et al. 1996. Long-term acceptance of skin and cardiac allografts after blocking CD40 and CD28 pathways. Nature 381: 434-438. https://doi.org/10.1038/381434a0
  33. Lin, H., R. Q. Wei, D. Gordon, P. Linsley, L. A. Turka, and S. F. Bolling. 1994. Review of CTLA4Ig use for allograft immunosuppression. Transplant. Proc. 26: 3200-3201.
  34. Ling, V., P. W. Wu, H. F. Finnerty, A. H. Sharpe, G. S. Gray, and M. Collins. 1999. Complete sequence determination of the mouse and human CTLA4 gene loci: Cross-species DNA sequence similarity beyond exon borders. Genomics 60: 341-355. https://doi.org/10.1006/geno.1999.5930
  35. Linsley, P. S., W. Brady, M. Urnes, L. S. Grosmaire, N. K. Damle, and J. A. Ledbetter. 1991. CTLA-4 is a second receptor for the B cell activation antigen B7. J. Exp. Med. 174: 561-569. https://doi.org/10.1084/jem.174.3.561
  36. Linsley, P. S. and J. A. Ledbetter. 1993. The role of the CD28 receptor during T cell responses to antigen. Annu. Rev. Immunol. 11: 191-212. https://doi.org/10.1146/annurev.iy.11.040193.001203
  37. Lorenz, H. M. and J. R. Kalden. 2002. Perspectives for TNFalpha-targeting therapies. Arthritis Res. 4 Suppl 3: S17-S24. https://doi.org/10.1186/ar564
  38. Malm, H., M. Corbascio, C. Osterholm, S. Cowan, C. P. Larsen, T. C. Pearson, and H. Ekberg. 2002. CTLA4Ig induces longterm graft survival of allogeneic skin grafts and totally inhibits T-cell proliferation in LFA-1-deficient mice. Transplantation 73: 293-297. https://doi.org/10.1097/00007890-200201270-00024
  39. Mastrangeli, R., E. Micangeli, and S. Donini. 1996. Cloning of murine LAG-3 by magnetic bead bound homologous probes and PCR (gene-capture PCR). Anal. Biochem. 241: 93-102. https://doi.org/10.1006/abio.1996.0382
  40. Moreland, L. W., R. Alten, F. Van den Bosch, T. Appelboom, M. Leon, P. Emery, et al. 2002. Costimulatory blockade in patients with rheumatoid arthritis: A pilot, dose-finding, doubleblind, placebo-controlled clinical trial evaluating CTLA-4Ig and LEA29Y eighty-five days after the first infusion. Arthritis Rheum. 46: 1470-1479. https://doi.org/10.1002/art.10294
  41. Murphy, W. J. and B. R. Blazar. 1999. New strategies for preventing graft-versus-host disease. Curr. Opin. Immunol. 11: 509-515. https://doi.org/10.1016/S0952-7915(99)00002-3
  42. Naparstek, Y. and P. H. Plotz. 1993. The role of autoantibodies in autoimmune disease. Annu. Rev. Immunol. 11: 79-104. https://doi.org/10.1146/annurev.iy.11.040193.000455
  43. Petric, R., D. Freeman, C. Wallace, J. McDonald, C. Stiller, and P. Keown. 1988. Effect of cyclosporine on urinary prostanoid excretion, renal blood flow, and glomerulotubular function. Transplantation 45: 883-889. https://doi.org/10.1097/00007890-198805000-00008
  44. Przepiorka, D., G. L. Phillips, V. Ratanatharathorn, M. Cottler-Fox, L. H. Sehn, J. H. Antin, et al. 1998. A phase II study of BTI-322, a monoclonal anti-CD2 antibody, for treatment of steroid-resistant acute graft-versus-host disease. Blood 92: 4066-4071.
  45. Reiter, Y. and I. Pastan. 1998. Recombinant Fv immunotoxins and Fv fragments as novel agents for cancer therapy and diagnosis. Trends Biotechnol. 16: 513-520. https://doi.org/10.1016/S0167-7799(98)01226-8
  46. Rodrigues, M., R. S. Nussenzweig, P. Romero, and F. Zavala. 1992. The in vivo cytotoxic activity of CD8+ T cell clones correlates with their levels of expression of adhesion molecules. J. Exp. Med. 175: 895-905. https://doi.org/10.1084/jem.175.4.895
  47. Schwartz, R. H. 1992. Costimulation of T lymphocytes: The role of CD28, CTLA-4, and B7/BB1 in interleukin-2 production and immunotherapy. Cell 71: 1065-1068. https://doi.org/10.1016/S0092-8674(05)80055-8
  48. Scott, D. L. 2005. Etanercept in arthritis. Int. J. Clin. Pract. 59: 114-118. https://doi.org/10.1111/j.1742-1241.2005.00380.x
  49. Storb, R., R. B. Epstein, T. C. Graham, and E. D. Thomas. 1970. Methotrexate regimens for control of graft-versus-host disease in dogs with allogeneic marrow grafts. Transplantation 9: 240-246. https://doi.org/10.1097/00007890-197003000-00007
  50. Subramanyam, M., G. Wands, R. Nabioullin, and M. A. Tepper. 1998. Soluble human lymphocyte activation gene-3 modulates allospecific T cell responses. Int. Immunol. 10: 679-689. https://doi.org/10.1093/intimm/10.5.679
  51. Sultan, P., J. S. Schechner, J. M. McNiff, P. S. Hochman, C. C. Hughes, M. I. Lorber, et al. 1997. Blockade of CD2-LFA-3 interactions protects human skin allografts in immunodeficient mouse/human chimeras. Nat. Biotechnol. 15: 759-762.
  52. Suthanthiran, M. and T. B. Strom. 1994. Renal transplantation. N. Engl. J. Med. 331: 365-376. https://doi.org/10.1056/NEJM199408113310606
  53. Triebel, F., S. Jitsukawa, E. Baixeras, S. Roman-Roman, C. Genevee, E. Viegas-Pequignot, and T. Hercend. 1990. LAG-3, a novel lymphocyte activation gene closely related to CD4. J. Exp. Med. 171: 1393-1405. https://doi.org/10.1084/jem.171.5.1393
  54. Via, C. S. and G. M. Shearer. 1988. T-cell interactions in autoimmunity: Insights from a murine model of graft-versushost disease. Immunol. Today 9: 207-213. https://doi.org/10.1016/0167-5699(88)91215-7
  55. Wallace, P. M., J. S. Johnson, J. F. MacMaster, K. A. Kennedy, P. Gladstone, and P. S. Linsley. 1994. CTLA4Ig treatment ameliorates the lethality of murine graft-versus-host disease across major histocompatibility complex barriers. Transplantation 58: 602-610. https://doi.org/10.1097/00007890-199409150-00013
  56. Wang, Q., M. Zhang, G. Ding, Y. Liu, Y. Sun, J. Wang, et al. 2003. Anti-ICAM-1 antibody and CTLA-4Ig synergistically enhance immature dendritic cells to induce donor-specific immune tolerance in vivo. Immunol. Lett. 90: 33-42. https://doi.org/10.1016/S0165-2478(03)00160-3
  57. Woodward, J. E., L. Qin, K. D. Chavin, J. Lin, T. Tono, Y. Ding, et al. 1996. Blockade of multiple costimulatory receptors induces hyporesponsiveness: Inhibition of CD2 plus CD28 pathways. Transplantation 62: 1011-1018. https://doi.org/10.1097/00007890-199610150-00021
  58. Wu, A. M., W. Chen, A. Raubitschek, L. E. Williams, M. Neumaier, R. Fischer, et al. 1996. Tumor localization of anti-CEA single-chain Fvs: Improved targeting by non-covalent dimers. Immunotechnology 2: 21-36. https://doi.org/10.1016/1380-2933(95)00027-5
  59. Wu, A. M., P. J. Yazaki, S. Tsai, K. Nguyen, A. L. Anderson, D. W. McCarthy, et al. 2000. High-resolution microPET imaging of carcinoembryonic antigen-positive xenografts by using a copper-64-labeled engineered antibody fragment. Proc. Natl. Acad. Sci. USA 97: 8495-8500. https://doi.org/10.1073/pnas.150228297
  60. Yamane, T., R. Yamamura, Y. Aoyama, H. Nakamae, T. Hasegawa, C. Sakamoto, et al. 2003. Infliximab for the treatment of severe steroid refractory acute graft-versus-host disease in three patients after allogeneic hematopoietic transplantation. Leuk. Lymphoma 44: 2095-2097. https://doi.org/10.1080/1042819031000123483
  61. Yim, S. B. and Y. H. Chung. 2004. Construction and production of concatameric human TNF receptor immunoglobulin fusion proteins. J. Microbiol. Biotechnol. 14: 81-89.
  62. Yin, D. and C. G. Fathman. 1995. Induction of tolerance to heart allografts in high responder rats by combining anti-CD4 with CTLA4Ig. J. Immunol. 155: 1655-1659.
  63. Yu, C., P. Linsley, K. Seidel, G. Sale, H. J. Deeg, R. A. Nash, and R. Storb. 2000. Cytotoxic T lymphocyte antigen 4-immunoglobulin fusion protein combined with methotrexate/cyclosporine as graft-versus-host disease prevention in a canine dog leukocyte antigen-nonidentical marrow transplant model. Transplantation 69: 450-454. https://doi.org/10.1097/00007890-200002150-00027