Transgenic expression of CTLA4-Ig by fetal pig neurons for xenotransplantation
Received: 06 September 2004 Accepted: 02 December 2004 DOI:
Cite this article as: Martin, C., Plat, M., Nerrière-Daguin, V. et al. Transgenic Res (2005) 14: 373. doi:10.1007/s11248-004-7268-4 Abstract
The transplantation of fetal porcine neurons is a potential therapeutic strategy for the treatment of human neurodegenerative disorders. A major obstacle to xenotransplantation, however, is the immune-mediated rejection that is resistant to conventional immunosuppression. To determine whether genetically modified donor pig neurons could be used to deliver immunosuppressive proteins locally in the brain, transgenic pigs were developed that express the human T cell inhibitory molecule hCTLA4-Ig under the control of the neuron-specific enolase promoter. Expression was found in various areas of the brain of transgenic pigs, including the mesencephalon, hippocampus and cortex. Neurons from 28-day old embryos secreted hCTLA4-Ig
in vitro and this resulted in a 50% reduction of the proliferative response of human T lymphocytes in xenogenic proliferation assays. Transgenic embryonic neurons also secreted hCTLA4-Ig and had developed normally in vivo several weeks after transplantation into the striatum of immunosuppressed rats that were used here to study the engraftment in the absence of immunity. In conclusion, these data show that neurons from our transgenic pigs express hCTLA4-Ig in situ and support the use of this material in future pre-clinical trials in neuron xenotransplantation. Keywords costimulation blockade immunosuppression neuron xenotransplantation
Caroline Martin,Martine Plat, Philippe Brachet, Bernard Vanhove - These authors have contributed equally to the development of this work.
References Bachoud-Levi, AC, Remy, P, Nguyen, JP, Brugieres, P, Lefaucheur, JP, Bourdet, C, et al. 2000 Motor and cognitive improvements in patients with Huntington’s disease after neural transplantation Lancet 356 1975 1979 CrossRef PubMed Barker, RA, Ratcliffe, E, McLaughlin, M, Richards, A, Dunnett, SB 2000 A role for complement in the rejection of porcine ventral mesencephalic xenografts in a rat model of Parkinson’s disease J Neurosci 20 3415 3424 PubMed Berghorn, KA, Bonnett, JH, Hoffman, GE 1994 cFos immunoreactivity is enhanced with biotin amplification J Histochem Cytochem 42 1635 1642 PubMed Bjorklund, A, Dunnett, SB, Brundin, P, Stoessl, AJ, Freed, CR, Breeze, RE, et al. 2003 Neural transplantation for the treatment of Parkinson’s disease Lancet Neurol 2 437 445 CrossRef PubMed Bolam, JP, Izzo, PN, Graybiel, AM 1988 Cellular substrate of the histochemically defined striosome/matrix system of the caudate nucleus: a combined Golgi and immunocytochemical study in cat and ferret Neuroscience 24 853 875 CrossRef PubMed Brevig, T, Holgersson, J, Widner, H 2000 Xenotransplantation for CNS repair: immunological barriers and strategies to overcome them Trends Neurosci 23 337 344 PubMed Brevig, T, Pedersen, EB, Kristensen, T, Zimmer, J 1997 Proliferative response of human T lymphocytes to porcine fetal brain cells Cell Transplant 6 571 577 CrossRef PubMed Cicchetti, F, Fodor, W, Deacon, TW, Horn, C, Rollins, S, Burton, W, et al. 2003 Immune parameters relevant to neural xenograft survival in the primate brain Xenotransplantation 10 41 49 CrossRef PubMed Croxford, JL, O’Neill, JK, Ali, RR, Browne, K, Byrnes, AP, Dallman, MJ, et al. 1998 Local gene therapy with CTLA4-immunoglobulin fusion protein in experimental allergic encephalomyelitis Eur J Immunol 28 3904 3916 CrossRef PubMed Deacon, T, Schumacher, J, Dinsmore, J, Thomas, C, Palmer, P, Kott, S, et al. 1997 Histological evidence of fetal pig neural cell survival after transplantation into a patient with Parkinson’s disease Nat Med 3 350 353 CrossRef PubMed Deacon, TW, Pakzaban, P, Burns, LH, Dinsmore, J, Isacson, O 1994 Cytoarchitectonic development, axon-glia relationships, and long distance axon growth of porcine striatal xenografts in rats Exp Neurol 130 151 167 CrossRef PubMed Edge, AS, Gosse, ME, Dinsmore, J 1998 Xenogeneic cell therapy: current progress and future developments in porcine cell transplantation Cell Transplant 7 525 539 CrossRef PubMed Felix, B, Leger, ME, Albe-Fessard, D, Marcilloux, JC, Rampin, O, Laplace, JP 1999 Stereotaxic atlas of the pig brain Brain Res Bull 49 1 137 CrossRef PubMed Fink, JS, Schumacher, JM, Ellias, SL, Palmer, EP, Saint-Hilaire, M, Shannon, K, et al. 2000 Porcine xenografts in Parkinson’s disease and Huntington’s disease patients: preliminary results Cell Transplant 9 273 278 PubMed Forss-Petter, S, Danielson, PE, Catsicas, S, Battenberg, E, Price, J, Nerenberg, M, et al. 1990 Transgenic mice expressing beta-galactosidase in mature neurons under neuron-specific enolase promoter control Neuron 5 187 197 CrossRef PubMed Galpern, WR, Burns, LH, Deacon, TW, Dinsmore, J, Isacson, O 1996 Xenotransplantation of porcine fetal ventral mesencephalon in a rat model of Parkinson’s disease: functional recovery and graft morphology Exp Neurol 140 1 13 CrossRef PubMed Garcia, AR, Deacon, TW, Dinsmore, J, Isacson, O 1995 Extensive axonal and glial fiber growth from fetal porcine cortical xenografts in the adult rat cortex Cell Transplant 4 515 527 CrossRef PubMed Grohmann, U, Orabona, C, Fallarino, F, Vacca, C, Calcinaro, F, Falorni, A, et al. 2002 CTLA-4-Ig regulates tryptophan catabolism in vivo Nat lmmunol 3 1097 1101 CrossRef Hoehn, M, Kustermann, E, Blunk, J, Wiedermann, D, Trapp, T, Wecker, S, et al. 2002 Monitoring of implanted stem cell migration in vivo: a highly resolved in vivo magnetic resonance imaging investigation of experimental stroke in rat Proc Natl Acad Sci USA 99 16267 16272 CrossRef PubMed Ideguchi, M, Kajiwara, K, Yoshikawa, K, Uchida, T, Ito, H 2000 Local adenovirus-mediated CTLA4-immunoglobulin expression suppresses the immune responses to adenovirus vectors in the brain Neuroscience 95 217 226 CrossRef PubMed Isacson, O, Deacon, T 1997 Neural transplantation studies reveal the brain’s capacity for continuous reconstruction Trends Neurosci 20 477 482 CrossRef PubMed Isacson, O, Deacon, TW, Pakzaban, P, Galpern, WR, Dinsmore, J, Burn, LH 1995 Transplanted xenogeneic neural cells in neurodegenerative disease models exhibit remarkable axonal target specificity and distinct growth patterns of glial and axonal fibres Nat Med 1 1189 1194 CrossRef PubMed Larsson, LC, Czech, KA, Brundin, P, Widner, H 2000 Intrastriatal ventral mesencephalic xenografts of porcine tissue in rats: immune responses and functional effects Cell Transplant 9 261 272 PubMed Lindvall, O, Brundin, P, Widner, H, Rehncrona, S, Gustavii, B, Frackowiak, R, et al. 1990 Grafts of fetal dopamine neurons survive and improve motor function in Parkinson’s disease Science 247 574 577 PubMed Linsley, PS, Brady, W, Urnes, M, Grosmaire, LS, Damle, NK, Ledbetter, JA 1991 CTLA-4 is a second receptor for the B cell activation antigen B7 J Exp Med 174 561 569 CrossRef PubMed Melchior, B, Remy, S, Nerriere-Daguin, V, Heslan, JM, Soulillou, JP, Brachet, P 2002 Temporal analysis of cytokine gene expression during infiltration of porcine neuronal grafts implanted into the rat brain J Neurosci Res 68 284 292 CrossRef PubMed Ménoret, S, Plat, M, Blancho, G, Martinat-Botté, F, Bernard, P, Karam, G, et al. 2004 Characterization of human CD55 and CD59 transgenic pigs and kidney xenotransplantation in the pig-to-baboon combination Transplantation 77 1468 1471 CrossRef PubMed Mittoux, V, Joseph, JM, Conde, F, Palfi, S, Dautry, C, Poyot, T, et al. 2000 Restoration of cognitive and motor functions by ciliary neurotrophic factor in a primate model of Huntington’s disease Hum Gene Ther 11 1177 1187 CrossRef PubMed Ni, HT, Merica, RR, Spellman, SR, Wang, JM, Low, WC 2000 Visualization of antigen-specific T cell activation in vivo in response to intracerebral administration of a xenopeptide Exp Neurol 164 362 370 CrossRef PubMed Okura, Y, Tanaka, R, Ono, K, Yoshida, S, Tanuma, N, Matsumoto, Y 1997 Treatment of rat hemiparkinson model with xenogeneic neural transplantation: tolerance induction by anti-T-cell antibodies J Neurosci Res 48 385 396 CrossRef PubMed Pages, FRM, Klasen, S, Battifora, M, Couez, D, Sweet, R, Truneh, A, Ward, SG, Olive, D 1996 Two distinct intracytoplasmic regions of the T-cell adhesion molecule CD28 participate in phosphatidylinositol 3-kinase association J Biol Chem 271 9403 9409 CrossRef PubMed Pakzaban, P, Deacon, TW, Burns, LH, Dinsmore, J, Isacson, O 1995 A novel mode of immunoprotection of neural xenotransplants: masking of donor major histocompatibility complex class I enhances transplant survival in the central nervous system Neuroscience 65 983 996 CrossRef PubMed Pedersen, EB, Zimmer, J, Finsen, B 1997 Triple immunosuppression protects murine intracerebral, hippocampal xenografts in adult rat hosts: effects on cellular infiltration, major histocompatibility complex antigen induction and blood-brain barrier leakage Neuroscience 78 685 701 CrossRef PubMed Peschanski, M, Bachoud-Levi, AC, Hantraye, P 2004 Integrating fetal neural transplants into a therapeutic strategy: the example of Huntington’s disease Brain 127 1219 1228 CrossRef PubMed Pursel, VG, Bolt, DJ, Miller, KF, Pinkert, CA, Hammer, RE, Palmiter, RD, et al. 1990 Expression and performance in transgenic pigs J Reprod Fertil Suppl 40 235 245 PubMed Remy, S, Canova, C, Daguin-Nerriere, V, Martin, C, Melchior, B, Neveu, I, et al. 2001 Different mechanisms mediate the rejection of porcine neurons and endothelial cells transplanted into the rat brain Xenotransplantation 8 136 148 CrossRef PubMed Sergent-Tanguy, S, Chagneau, C, Neveu, I, Naveilhan, P 2003 Fluorescent activated cell sorting (FACS): a rapid and reliable method to estimate the number of neurons in a mixed population J Neurosci Methods 129 73 79 CrossRef PubMed Tsuji, A, Tamai, I, Sakata, A, Tenda, Y, Terasaki, T 1993 Restricted transport of cyclosporin A across the blood-brain barrier by a multidrug transporter, P-glycoprotein Biochem Pharmacol 46 1096 1099 CrossRef PubMed Twyman, RM, Jones, EA 1997 Sequences in the proximal 5′ flanking region of the rat neuron-specific enolase (NSE) gene are sufficient for cell type-specific reporter gene expression J Mol Neurosci 8 63 73 PubMed Uchida, T, Kajiwara, K, Ideguchi, M, Yoshikawa, K, Morioka, J, Suzuki, M 2001 Co-administration of adenovirus vector expressing CTLA4-Ig prolongs transgene expression in the brain of mice sensitized with adenovirus Brain Res 898 272 280 CrossRef PubMed Wallace, PM, Johnson, JS, MacMaster, JF, Kennedy, KA, Gladstone, P, Linsley, PS 1994 CTLA4Ig treatment ameliorates the lethality of murine graft-versus-host disease across major histocompatibility complex barriers Transplantation 58 602 610 PubMed Wallace, PM, Rodgers, JN, Leytze, GM, Johnson, JS, Linsley, PS 1995 Induction and reversal of long-lived specific unresponsiveness to a T-dependent antigen following CTLA4Ig treatment J Immunol 154 5885 5895 PubMed Wenkel, H, Streilein, JW, Young, MJ 2000 Systemic immune deviation in the brain that does not depend on the integrity of the blood-brain barrier J Immunol l 64 5125 5131 Wood, MJ, Sloan, DJ, Wood, KJ, Charlton, HM 1996 Indefinite survival of neural xenografts induced with anti-CD4 monoclonal antibodies Neuroscience 70 775 789 CrossRef PubMed