Background Transplantation of embryonic pig pancreatic tissue as a source of insulin has been suggested for the cure of diabetes. However, previous limited clinical trials failed in their attempts to treat diabetic patients by transplantation of advanced gestational age porcine embryonic pancreas. In the present study we examined growth potential, functionality, and immunogenicity of pig embryonic pancreatic tissue harvested at different gestational ages. Methods and Findings Implantation of embryonic pig pancreatic tissues of different gestational ages in SCID mice reveals that embryonic day 42 (E42) pig pancreas can enable a massive growth of pig islets for prolonged periods and restore normoglycemia in diabetic mice. Furthermore, both direct and indirect T cell rejection responses to the xenogeneic tissue demonstrated that E42 tissue, in comparison to E56 or later embryonic tissues, exhibits markedly reduced immunogenicity. Finally, fully immunocompetent diabetic mice grafted with the E42 pig pancreatic tissue and treated with an immunosuppression protocol comprising CTLA4-Ig and anti–CD40 ligand (anti-CD40L) attained normal blood glucose levels, eliminating the need for insulin. Conclusions These results emphasize the importance of selecting embryonic tissue of the correct gestational age for optimal growth and function and for reduced immunogenicity, and provide a proof of principle for the therapeutic potential of E42 embryonic pig pancreatic tissue transplantation in diabetes.
References
[1]
King H, Aubert RE, Herman WH (1998) Global burden of diabetes, 1995–2025: Prevalence, numerical estimates, and projections. Diabetes Care 21: 1414–1431.
[2]
Dunstan DW, Zimmet PZ, Welborn TA, De Courten MP, Cameron AJ, et al. (2002) The rising prevalence of diabetes and impaired glucose tolerance: The Australian Diabetes, Obesity and Lifestyle Study. Diabetes Care 25: 829–834.
[3]
UK Prospective Diabetes Study (UKPDS) Group (1998) Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UKPDS 33). Lancet 352: 837–853.
[4]
Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications Research Group (2000) Retinopathy and nephropathy in patients with type 1 diabetes four years after a trial of intensive therapy. N Engl J Med 342: 381–389.
[5]
White NH, Cleary PA, Dahms W, Goldstein D, Malone J, et al. (2001) Beneficial effects of intensive therapy of diabetes during adolescence. J Pediatr 139: 804–812.
[6]
Nathan DM, Cleary PA, Backlund JY, Genuth SM, Lachin JM, et al. (2005) Intensive diabetes treatment and cardiovascular disease in patients with type 1 diabetes. N Engl J Med 353: 2643–2653.
[7]
Shapira Z, Yussim A, Mor E (1999) Pancreas transplantation. J Pediatr Endocrinol Metab 12: 3–15.
[8]
Shapiro AM, Lakey JR, Ryan EA, Korbutt GS, Toth E, et al. (2000) Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N Engl J Med 343: 230–238.
[9]
Sutherland DE, Gruessner RW, Dunn DL, Matas AJ, Humar A, et al. (2001) Lessons learned from more than 1,000 pancreas transplants at a single institution. Ann Surg 233: 463–501.
[10]
Ryan EA, Lakey JR, Rajotte RV, Korbutt GS, Kin T, et al. (2001) Clinical outcomes and insulin secretion after islet transplantation with the Edmonton protocol. Diabetes 50: 710–719.
[11]
Ryan EA, Lakey JR, Paty BW, Imes S, Korbutt GS, et al. (2002) Successful islet transplantation: Continued insulin reserve provides long-term glycemic control. Diabetes 51: 2148–2157.
[12]
Robertson RP (2004) Islet transplantation as a treatment for diabetes—A work in progress. N Engl J Med 350: 694–705.
[13]
Hering BJ, Kandaswamy R, Ansite JD, Eckman PM, Nakano M, et al. (2005) Single-donor, marginal-dose islet transplantation in patients with type 1 diabetes. JAMA 293: 830–835.
[14]
Danovitch GM, Cohen DJ, Weir MR, Stock PG, Bennett WM, et al. (2005) Current status of kidney and pancreas transplantation in the United States, 1994–2003. Am J Transplant 5: 904–915.
[15]
Wynn JJ, Distant DA, Pirsch JD, Norman D, Gaber AO, et al. (2004) Kidney and pancreas transplantation. Am J Transplant 4: 72–80.
[16]
Ojo AO, Heinrichs D, Emond JC, McGowan JJ, Guidinger MK, et al. (2004) Organ donation and utilization in the USA. Am J Transplant 4: 27–37.
[17]
Abouna GM (2003) Ethical issues in organ and tissue transplantation. Exp Clin Transplant 1: 125–138.
[18]
Elliott RB, Escobar L, Garkavenko O, Croxson MC, Schroeder BA, et al. (2000) No evidence of infection with porcine endogenous retrovirus in recipients of encapsulated porcine islet xenografts. Cell Transplant 9: 895–901.
[19]
Langford GA, Galbraith D, Whittam AJ, McEwan P, Fernandez-Suarez XM, et al. (2001) In vivo analysis of porcine endogenous retrovirus expression in transgenic pigs. Transplantation 72: 1996–2000.
[20]
Tacke SJ, Bodusch K, Berg A, Denner J (2001) Sensitive and specific immunological detection methods for porcine endogenous retroviruses applicable to experimental and clinical xenotransplantation. Xenotransplantation 8: 125–135.
[21]
Paradis K, Langford G, Long Z, Heneine W, Sandstrom P, et al. (1999) Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue. The XEN 111 Study Group. Science 285: 1236–1241.
[22]
Clark DA, Fryer JF, Tucker AW, McArdle PD, Hughes AE, et al. (2003) Porcine cytomegalovirus in pigs being bred for xenograft organs: Progress towards control. Xenotransplantation 10: 142–148.
[23]
Ogata K, Platt JL (2004) Potential applications and prospects for cardiac xenotransplantation. J Heart Lung Transplant 23: 515–526.
[24]
Foglia RP, DiPreta J, Statter MB, Donahoe PK (1986) Fetal allograft survival in immunocompetent recipients is age dependent and organ specific. Ann Surg 204: 402–410.
[25]
Statter MB, Foglia RP, Parks DE, Donahoe PK (1988) Fetal and postnatal testis shows immunoprivilege as donor tissue. J Urol 139: 204–210.
[26]
Dekel B, Burakova T, Ben-Hur H, Marcus H, Oren R, et al. (1997) Engraftment of human kidney tissue in rat radiation chimera: II. Human fetal kidneys display reduced immunogenicity to adoptively transferred human peripheral blood mononuclear cells and exhibit rapid growth and development. Transplantation 64: 1550–1558.
[27]
Eventov-Friedman S, Katchman H, Shezen E, Aronovich A, Tchorsh D, et al. (2005) Embryonic pig liver, pancreas, and lung as a source for transplantation: Optimal organogenesis without teratoma depends on distinct time windows. Proc Natl Acad Sci U S A 102: 2928–2933.
[28]
Bouwens L (1998) Cytokeratins and cell differentiation in the pancreas. J Pathol 184: 234–239.
[29]
Dekel B, Burakova T, Arditti FD, Reich-Zeliger S, Milstein O, et al. (2003) Human and porcine early kidney precursors as a new source for transplantation. Nat Med 9: 53–60.
[30]
Castaing M, Peault B, Basmaciogullari A, Casal I, Czernichow P, et al. (2001) Blood glucose normalization upon transplantation of human embryonic pancreas into beta-cell-deficient SCID mice. Diabetologia 44: 2066–2076.
[31]
Eizirik DL, Pipeleers DG, Ling Z, Welsh N, Hellerstrom C, et al. (1994) Major species differences between humans and rodents in the susceptibility to pancreatic beta-cell injury. Proc Natl Acad Sci U S A 91: 9253–9256.
[32]
Tyrberg B, Andersson A, Borg LA (2001) Species differences in susceptibility of transplanted and cultured pancreatic islets to the beta-cell toxin alloxan. Gen Comp Endocrinol 122: 238–251.
[33]
Sayegh MH, Turka LA (1998) The role of T-cell costimulatory activation pathways in transplant rejection. N Engl J Med 338: 1813–1821.
[34]
Lechler R, Ng WF, Steinman RM (2001) Dendritic cells in transplantation—Friend or foe? Immunity 14: 357–368.
[35]
Elwood ET, Larsen CP, Cho HR, Corbascio M, Ritchie SC, et al. (1998) Prolonged acceptance of concordant and discordant xenografts with combined CD40 and CD28 pathway blockade. Transplantation 65: 1422–1428.
[36]
Tadaki DK, Craighead N, Saini A, Celniker A, Burkly LC, et al. (2000) Costimulatory molecules are active in the human xenoreactive T-cell response but not in natural killer-mediated cytotoxicity. Transplantation 70: 162–167.
[37]
Groth CG, Korsgren O, Tibell A, Tollemar J, Moller E, et al. (1994) Transplantation of porcine fetal pancreas to diabetic patients. Lancet 344: 1402–1404.
[38]
Rogers SA, Chen F, Talcott M, Hammerman MR (2004) Islet cell engraftment and control of diabetes in rats after transplantation of pig pancreatic anlagen. Am J Physiol Endocrinol Metab 286: E502–E509.
[39]
Rogers SA, Liapis H, Hammerman MR (2005) Normalization of glucose post-transplantation of pig pancreatic anlagen into non-immunosuppressed diabetic rats depends on obtaining anlagen prior to embryonic day 35. Transpl Immunol 14: 67–75.
[40]
Groth CG, Tibell A, Wennberg L, Korsgren O (1999) Xenoislet transplantation: Experimental and clinical aspects. J Mol Med 77: 153–154.
[41]
Tu J, Khoury P, Williams L, Tuch BE (2004) Comparison of fetal porcine aggregates of purified beta-cells versus islet-like cell clusters as a treatment of diabetes. Cell Transplant 13: 525–534.
[42]
Dekel B, Burakova T, Marcus H, Shezen E, Polack S, et al. (1997) Engraftment of human kidney tissue in rat radiation chimera: I. A new model of human kidney allograft rejection. Transplantation 64: 1541–1550.
[43]
Korsgren O, Andersson A, Sandler S (1993) Pretreatment of fetal porcine pancreas in culture with nicotinamide accelerates reversal of diabetes after transplantation to nude mice. Surgery 113: 205–214.
[44]
Jiang FX, Cram DS, DeAizpurua HJ, Harrison LC (1999) Laminin-1 promotes differentiation of fetal mouse pancreatic beta-cells. Diabetes 48: 722–730.
[45]
Miettinen PJ, Huotari M, Koivisto T, Ustinov J, Palgi J, et al. (2000) Impaired migration and delayed differentiation of pancreatic islet cells in mice lacking EGF-receptors. Development 127: 2617–2627.
[46]
Bhushan A, Itoh N, Kato S, Thiery JP, Czernichow P, et al. (2001) Fgf10 is essential for maintaining the proliferative capacity of epithelial progenitor cells during early pancreatic organogenesis. Development 128: 5109–5117.
[47]
Cras-Meneur C, Elghazi L, Czernichow P, Scharfmann R (2001) Epidermal growth factor increases undifferentiated pancreatic embryonic cells in vitro: A balance between proliferation and differentiation. Diabetes 50: 1571–1579.
[48]
Lammert E, Cleaver O, Melton D (2001) Induction of pancreatic differentiation by signals from blood vessels. Science 294: 564–567.
[49]
Elghazi L, Cras-Meneur C, Czernichow P, Scharfmann R (2002) Role for FGFR2IIIb-mediated signals in controlling pancreatic endocrine progenitor cell proliferation. Proc Natl Acad Sci U S A 99: 3884–3889.
[50]
Movassat J, Beattie GM, Lopez AD, Portha B, Hayek A (2003) Keratinocyte growth factor and beta-cell differentiation in human fetal pancreatic endocrine precursor cells. Diabetologia 46: 822–829.
[51]
Miralles F, Czernichow P, Scharfmann R (1998) Follistatin regulates the relative proportions of endocrine versus exocrine tissue during pancreatic development. Development 125: 1017–1024.
[52]
Si Z, Tuch BE, Walsh DA (2001) Development of human fetal pancreas after transplantation into SCID mice. Cells Tissues Organs 168: 147–157.
[53]
Castaing M, Duvillie B, Quemeneur E, Basmaciogullari A, Scharfmann R (2005) Ex vivo analysis of acinar and endocrine cell development in the human embryonic pancreas. Dev Dyn 234: 339–345.
[54]
Desai DM, Adams GA, Wang X, Alfrey EJ, Sibley RK, et al. (1999) The influence of combined trophic factors on the success of fetal pancreas grafts. Transplantation 68: 491–496.
[55]
Otonkoski T, Ustinov J, Rasilainen S, Kallio E, Korsgren O, et al. (1999) Differentiation and maturation of porcine fetal islet cells in vitro and after transplantation. Transplantation 68: 1674–1683.
[56]
Medawar P (1953) Some immunological and endocrinological problems raised by the evolution of viviparity in vertebrates. Symp Soc Exp Biol 7: 320–323.
[57]
Koulmanda M, Laufer TM, Auchincloss H, Smith RN (2004) Prolonged survival of fetal pig islet xenografts in mice lacking the capacity for an indirect response. Xenotransplantation 11: 525–530.
[58]
Mirenda V, Golshayan D, Read J, Berton I, Warrens AN, et al. (2005) Achieving permanent survival of islet xenografts by independent manipulation of direct and indirect T-cell responses. Diabetes 54: 1048–1055.
[59]
Lehnert AM, Yi S, Burgess JS, O'Connell PJ (2000) Pancreatic islet xenograft tolerance after short-term costimulation blockade is associated with increased CD4+ T cell apoptosis but not immune deviation. Transplantation 69: 1176–1185.
[60]
Samstein B, Platt JL (2001) Xenotransplantation and tolerance. Philos Trans R Soc Lond B Biol Sci 356: 749–758.
[61]
Hering BJ, Wijkstrom M, Graham ML, Hardstedt M, Aasheim TC, et al. (2006) Prolonged diabetes reversal after intraportal xenotransplantation of wild-type porcine islets in immunosuppressed nonhuman primates. Nat Med 2: 301–303.
[62]
Cardona K, Korbutt GS, Milas Z, Lyon J, Cano J, et al. (2006) Long-term survival of neonatal porcine islets in nonhuman primates by targeting costimulation pathways. Nat Med 12: 304–306.
[63]
Kawai T, Andrews D, Colvin RB, Sachs DH, Cosimi AB (2000) Thromboembolic complications after treatment with monoclonal antibody against CD40 ligand. Nat Med 6: 114.
[64]
Schuler W, Bigaud M, Brinkmann V, Di Padova F, Geisse S, et al. (2004) Efficacy and safety of ABI793, a novel human anti-human CD154 monoclonal antibody, in cynomolgus monkey renal allotransplantation. Transplantation 77: 717–726.
