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http://dx.doi.org/10.4014/jmb.1201.01026

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)
Publication Information
Journal of Microbiology and Biotechnology / v.22, no.8, 2012 , pp. 1066-1076 More about this Journal
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.
Keywords
Cytotoxic T lymphocyte antigen-4; lymphocyte activation gene-3; tetravalent immunoadhesin; graft-versus-host disease; mixed lymphocyte reaction;
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1 Lafferty, K. J. 1994. A contemporary view of transplantation tolerance: An immunologist's perspective. Clin. Transplant. 8: 181-187.
2 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.   DOI   ScienceOn
3 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.
4 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.   DOI   ScienceOn
5 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.   DOI   ScienceOn
6 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.   DOI   ScienceOn
7 Lorenz, H. M. and J. R. Kalden. 2002. Perspectives for TNFalpha-targeting therapies. Arthritis Res. 4 Suppl 3: S17-S24.   DOI
8 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.   DOI   ScienceOn
9 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.   DOI   ScienceOn
10 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.   DOI   ScienceOn
11 Murphy, W. J. and B. R. Blazar. 1999. New strategies for preventing graft-versus-host disease. Curr. Opin. Immunol. 11: 509-515.   DOI   ScienceOn
12 Naparstek, Y. and P. H. Plotz. 1993. The role of autoantibodies in autoimmune disease. Annu. Rev. Immunol. 11: 79-104.   DOI   ScienceOn
13 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.   DOI   ScienceOn
14 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.
15 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.   DOI   ScienceOn
16 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.   DOI   ScienceOn
17 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.   DOI   ScienceOn
18 Scott, D. L. 2005. Etanercept in arthritis. Int. J. Clin. Pract. 59: 114-118.   DOI   ScienceOn
19 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.   DOI   ScienceOn
20 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.   DOI   ScienceOn
21 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.
22 Suthanthiran, M. and T. B. Strom. 1994. Renal transplantation. N. Engl. J. Med. 331: 365-376.   DOI   ScienceOn
23 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.   DOI   ScienceOn
24 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.   DOI   ScienceOn
25 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.   DOI   ScienceOn
26 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.   DOI   ScienceOn
27 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.   DOI   ScienceOn
28 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.   DOI   ScienceOn
29 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.   DOI   ScienceOn
30 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.   DOI   ScienceOn
31 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.
32 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.
33 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.   DOI   ScienceOn
34 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.   DOI   ScienceOn
35 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.   DOI   ScienceOn
36 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.   DOI   ScienceOn
37 Antin, J. H. and J. L. Ferrara. 1992. Cytokine dysregulation and acute graft-versus-host disease. Blood 80: 2964-2968.
38 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.   DOI   ScienceOn
39 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.
40 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.
41 Bluestone, J. A., E. W. St Clair, and L. A. Turka. 2006. CTLA4Ig: Bridging the basic immunology with clinical application. Immunity 24: 233-238.   DOI   ScienceOn
42 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.   DOI   ScienceOn
43 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.   DOI   ScienceOn
44 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.
45 Emery, P. 2003. The therapeutic potential of costimulatory blockade with CTLA4Ig in rheumatoid arthritis. Expert Opin. Investig. Drugs 12: 673-681.   DOI   ScienceOn
46 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.   DOI   ScienceOn
47 Guidotti, L. G. and F. V. Chisari. 2000. Cytokine-mediated control of viral infections. Virology 273: 221-227.   DOI   ScienceOn
48 Gottlieb, A. B. 2004. Etanercept for the treatment of psoriasis and psoriatic arthritis. Dermatol. Ther. 17: 401-408.   DOI   ScienceOn
49 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.   DOI   ScienceOn
50 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.   DOI   ScienceOn
51 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.
52 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.
53 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.   DOI   ScienceOn
54 Hanahan, D. 1983. Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166: 557-580.   DOI
55 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.   DOI   ScienceOn
56 Jacobsohn, D. A. and G. B. Vogelsang. 2002. Novel pharmacotherapeutic approaches to prevention and treatment of GVHD. Drugs 62: 879-889.   DOI   ScienceOn
57 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.   DOI   ScienceOn
58 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.   DOI   ScienceOn
59 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.   DOI   ScienceOn
60 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.   DOI   ScienceOn
61 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.   DOI   ScienceOn
62 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.   DOI   ScienceOn
63 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.   DOI   ScienceOn