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Vol. 25, No. 1, 2009
Issue release date: August 2009
Section title: Original Paper
Free Access
Fetal Diagn Ther 2009;25:102–110
(DOI:10.1159/000203399)

Immune Ontogeny and Engraftment Receptivity in the Sheep Fetus

Skopal-Chase J.L.a · Pixley J.S.b · Torabi A.a · Cenariu M.C.a · Bhat A.b · Thain D.S.a · Frederick N.M.a · Groza D.M.a · Zanjani E.D.a
aDepartment of Animal Biotechnology, University of Nevada, and bDivision of Rheumatology/Immunology, Department of Medicine, VA Sierra Health Care System and University of Nevada School of Medicine, Reno, Nev., USA
email Corresponding Author

Abstract

Objective: The biologic explanation for fetal receptivity to donor engraftment and subsequent long-term tolerance following transplantation early in gestation is not known. We investigated the role fetal immune ontogeny might play in fetal transplantation tolerance in sheep. Methods: Engraftmentof allogeneic and xenogeneicHSC was determined 60 days following transplantation at different time points in sheep fetal gestation. Parallel analysis of surface differentiation antigen expression on cells from lymphoid organs of timed gestational age fetal sheep was determined by flow cytometry using available reagents. Results: An engraftment window was identified after day 52 gestation lasting until day 71 (term gestation: 145 days). This period was associated with the expression of the leukocyte common antigen CD45 on all cells in the thymus. Double-positive and single-positive CD4 and CD8 cells began appearing in the thymus just prior (day 45 gestation) to the beginning of the engraftment window, while single-positive CD4 or CD8 cells do not begin appearing in peripheral organs until late in the engraftment period, suggesting deletional mechanisms may be operative. In concert, surface IgM-positive cells express CD45 in the thymus at day 45, with a comparable delay in the appearance of IgM/CD45 cells in the periphery until late in the engraftment window. Conclusions: These findings support a central role for the thymus in multilineage immune cell maturation during the period of fetal transplantation receptivity. Further, they suggest that fetal engraftment receptivity is due to gestational age-dependent deletional tolerance.

© 2009 S. Karger AG, Basel


  

Key Words

  • Fetal transplantation tolerance
  • Fetal immune ontogeny
  • Deletional tolerance

References

  1. Pixley JS, Mackintosh FR, Zanjani ED: Experimental and clinical basis of intrauterine stem cell transplantation. Rev Clin Exp Hematol 1999;8:11–32.
  2. Zanjani ED, Almeida-Porada G, Livingston AG, Zeng H, Ogawa M: Reversible expression of CD34 by adult human bone marrow long-term engrafting hematopoietic stem cells. Exp Hematol 2003;31:406–412.
  3. Zanjani ED: The human sheep xenograft model for the study of the in vivo potential of human HSC and in utero gene transfer. Stem Cells 2000;18:151.
  4. Almeida-Porada G, Porada C, Gupta N, Torabi A, Thain D, Zanjani ED: The human-sheep chimeras as a model for human stem cell mobilization and evaluation of hematopoietic grafts’ potential. Exp Hematol 2007;35:1594–1600.
  5. Porada CD, Tran N, Eglitis M, Moen RC, Troutman L, Flake AW, Zhao Y, Anderson WF, Zanjani ED: In utero gene therapy: transfer and long-term expression of the bacterial neo(r) gene in sheep after direct injection of retroviral vectors into preimmune fetuses. Hum Gene Ther 1998;9:1571–1585.
  6. Billingham RE, Brent L, Medawar PB: Actively acquired tolerance of foreign cells. Nature 1953;172:603–606.
  7. Flake AW, Harrison MR, Adzick NS, Zanjani ED: Transplantation of fetal hematopoietic stem cells in utero: the creation of hematopoietic chimeras. Science 1986;233:776–778.
  8. Zanjani ED, Pallavicini MG, Ascensao JL, Flake AW, Langlois RG, Reitsma M, Mackintosh FR, Stutes D, Harrison MR, Tavassoli M: Engraftment and long-term expression of human fetal hemopoietic stem cells in sheep following transplantation in utero. J Clin Invest 1992;89:1178–1188.
  9. Silverstein A, Prendergast RA, Kraner K: Fetal response to antigenic stimulus. IV. Rejection of skin homografts by the fetal lamb. J Exp Med 1964;119:955–964.
  10. Tran ND, Porada CD, Almeida-Porada G, Glimp HA, Anderson WF, Zanjani ED: Induction of stable prenatal tolerance to β- galactosidase by in utero gene transfer into preimmune sheep fetuses. Blood 2001;97:3417–3423.
  11. Ohki H, Martin C, Corbel C, Coltey M, Le Douarin NM: Tolerance induced by thymic epithelial grafts in birds. Science 1987;237:1032–1035.
  12. Lutzko C, Meertens L, Li L, Zhao Y, Abrams-Ogg A, Woods JP, Kruth S, Hough MR, Dube ID: Human hematopoietic progenitors engraft in fetal canine recipients and expand with neonatal injection of fibroblasts expressing human hematopoietic cytokines. Exp Hematol 2002;30:801–808.
  13. Lee PW, Cina RA, Randolph MA, Goodrich J, Rowland H, Arellano R, Kim HB, Sachs DH, Huang CA: Stable multilineage chimerism across full MHC barriers without graft-versus-host disease following in utero bone marrow transplantation in pigs. Exp Hematol 2005;33:371–379.
  14. Peranteau WH, Hayashi S, Hsieh M, Shaaban AF, Flake AW: High-level allogeneic chimerism achieved by prenatal tolerance induction and postnatal non-myeloablative bone marrow transplantation. Blood 2002;100:2225–2234.
  15. Crombleholme TM, Harrison MR, Zanjani ED: In utero transplantation of hematopoietic stem cells in sheep: the role of T cells in engraftment and graft-versus-host disease. J Pediatr Surg 1990;25:885–892.
  16. Pixley JS, Zanjani ED, Shaft DM, Porada C, Mackintosh FR: Prolonged hematopoietic chimerism in normal mice transplanted in utero with human hematopoietic stem cells. Pathobiology 1998;66:230–239.
  17. Carlson BM: Fetal period and birth; in Human Embryology and Development. New York, Churchill Livingstone, 1999, pp 447–468.
  18. Maddox JF, Mackay CR, Brandon MR: Ontogeny of ovine lymphocytes. I. An immunohistological study on the development of T lymphocytes in the sheep embryo and fetal thymus. Immunology 1987;62:97–105.
  19. Owen RD: Immunogenetic consequences of vascular anastomoses between bovine twins. Science 1945;102:400–401.
  20. Cancro MP: Crossing the threshold of lymphocyte activation. J Immunol 2005;174:5159–5160.
  21. Schwartz RH, Mueller DL: Immunological tolerance; in Paul WE (ed): Fundamental Immunology. New York, Lippincott-Raven Press, 1999, pp 901–963.
  22. Hermiston ML, Xu Z, Weiss A: CD45: a critical regulator of signaling thresholds in immune cells. Annu Rev Immunol 2003;21:107–137.
  23. Maddox JF, Mackay CR, Brandon MR: The sheep analogue of leucocyte common antigen. Immunology 1985;55:347–353.
  24. Flake AW, Zanjani ED: In utero hematopoietic stem cell transplantation: ontogenic opportunities and biologic barriers. Blood 1999;94:2179–2191.
  25. Laufer TM, Glimcher LH, Lo D: Using thymus anatomy to dissect T-cell repertoire selection. Semin Immunol 1999;11:65–70.
  26. Naquet P, Naspetti M, Boyd R: Development, organization and function of the thymic medulla in normal, immunodeficient or autoimmune mice. Semin Immunol 1999;11:47–55.
  27. Trop S, Charron J, Arguin C, Lesage S, Hugo P: Thymic selection generates T cells expressing self-reactive TCRs in the absence of CD45. J Immunol 2000;165:3073–3079.
  28. Nemazee DA, Burki K: Clonal deletion of B lymphocytes in a transgenic mouse bearing anti-MHC class I antibody genes. Nature 1989;337:562–566.
  29. Rodriguez-Pinto D: B cells as antigen presenting cells. Cell Immunol 2005;238:67–75.
  30. Roux E, Dumont-Girard F, Starobinski M, Siegrist CA, Helg C, Chapuis B, Roosnek E: Recovery of immune reactivity after T-cell-depleted bone marrow transplantation depends on thymic activity. Blood 2000;96:2299–2303.
  31. De Kleer I, Vastert B, Klein M, Teklenburg G, Arkesteijn G, Yung GP, Albani S, Kuis W, Wulffraat N, Prakken B: Autologous stem cell transplantation for autoimmunity induces immunologic self-tolerance by reprogramming autoreactive T cells and restoring the CD4+CD25+ immune regulatory network. Blood 2006;107:1696–1702.
  32. Wardemann H, Nussenzweig MC: B-cell self-tolerance in humans. Adv Immunol 2007;95:83–110.
  33. Singh NJ, Schwartz RH: Primer: mechanisms of immunologic tolerance. Nat Clin Pract Rheumatol 2006;2:44–52.
  34. Carrier E, Lee TH, Busch MP, Cowan MJ: Induction of tolerance in nondefective mice after in utero transplantation of major histocompatibility complex-mismatched fetal hematopoietic stem cells. Blood 1995;86:4681–4690.
  35. Kim HB, Shaaban AF, Milner R, Fichter C, Flake AW: In utero bone marrow transplantation induces donor-specific tolerance by a combination of clonal deletion and clonal anergy. J Pediatr Surg 1999;34:726–729.
  36. Sefrioui H, Donahue J, Srivastava AS, Gilpin E, Lee TH, Carrier E: Alloreactivity following in utero transplantation of cytokine-stimulated hematopoietic stem cells: the role of recipient CD4– cells. Exp Hematol 2002;30:617–624.
  37. Fairchild PJ, Waldmann H: Extrathymic signals regulate the onset of T-cell repertoire selection. Eur J Immunol 2000;30:1948–1956.
  38. Larsen WJ: Development of the head and neck; in Human Embryology. Cincinnati, Churchill Livingstone, 1997, pp 347–374.
  39. Stites DP, Pavia CS: Ontogeny of human T cells. Pediatrics 1979;64:795–802.

  

Author Contacts

John S. Pixley, MD
VAMC, 1000 Locust Street
Reno, NV 89502-2597 (USA)
Tel. +1 775 328 1430, Fax +1 775 328 1769
E-Mail jpixley@medicine.nevada.edu

  

Article Information

Received: December 18, 2007
Accepted after revision: May 30, 2008
Published online: February 25, 2009
Number of Print Pages : 9
Number of Figures : 6, Number of Tables : 0, Number of References : 39

  

Publication Details

Fetal Diagnosis and Therapy (Clinical Advances and Basic Research)

Vol. 25, No. 1, Year 2009 (Cover Date: August 2009)

Journal Editor: Gratacós E. (Barcelona)
ISSN: 1015-3837 (Print), eISSN: 1421-9964 (Online)

For additional information: http://www.karger.com/FDT


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References

  1. Pixley JS, Mackintosh FR, Zanjani ED: Experimental and clinical basis of intrauterine stem cell transplantation. Rev Clin Exp Hematol 1999;8:11–32.
  2. Zanjani ED, Almeida-Porada G, Livingston AG, Zeng H, Ogawa M: Reversible expression of CD34 by adult human bone marrow long-term engrafting hematopoietic stem cells. Exp Hematol 2003;31:406–412.
  3. Zanjani ED: The human sheep xenograft model for the study of the in vivo potential of human HSC and in utero gene transfer. Stem Cells 2000;18:151.
  4. Almeida-Porada G, Porada C, Gupta N, Torabi A, Thain D, Zanjani ED: The human-sheep chimeras as a model for human stem cell mobilization and evaluation of hematopoietic grafts’ potential. Exp Hematol 2007;35:1594–1600.
  5. Porada CD, Tran N, Eglitis M, Moen RC, Troutman L, Flake AW, Zhao Y, Anderson WF, Zanjani ED: In utero gene therapy: transfer and long-term expression of the bacterial neo(r) gene in sheep after direct injection of retroviral vectors into preimmune fetuses. Hum Gene Ther 1998;9:1571–1585.
  6. Billingham RE, Brent L, Medawar PB: Actively acquired tolerance of foreign cells. Nature 1953;172:603–606.
  7. Flake AW, Harrison MR, Adzick NS, Zanjani ED: Transplantation of fetal hematopoietic stem cells in utero: the creation of hematopoietic chimeras. Science 1986;233:776–778.
  8. Zanjani ED, Pallavicini MG, Ascensao JL, Flake AW, Langlois RG, Reitsma M, Mackintosh FR, Stutes D, Harrison MR, Tavassoli M: Engraftment and long-term expression of human fetal hemopoietic stem cells in sheep following transplantation in utero. J Clin Invest 1992;89:1178–1188.
  9. Silverstein A, Prendergast RA, Kraner K: Fetal response to antigenic stimulus. IV. Rejection of skin homografts by the fetal lamb. J Exp Med 1964;119:955–964.
  10. Tran ND, Porada CD, Almeida-Porada G, Glimp HA, Anderson WF, Zanjani ED: Induction of stable prenatal tolerance to β- galactosidase by in utero gene transfer into preimmune sheep fetuses. Blood 2001;97:3417–3423.
  11. Ohki H, Martin C, Corbel C, Coltey M, Le Douarin NM: Tolerance induced by thymic epithelial grafts in birds. Science 1987;237:1032–1035.
  12. Lutzko C, Meertens L, Li L, Zhao Y, Abrams-Ogg A, Woods JP, Kruth S, Hough MR, Dube ID: Human hematopoietic progenitors engraft in fetal canine recipients and expand with neonatal injection of fibroblasts expressing human hematopoietic cytokines. Exp Hematol 2002;30:801–808.
  13. Lee PW, Cina RA, Randolph MA, Goodrich J, Rowland H, Arellano R, Kim HB, Sachs DH, Huang CA: Stable multilineage chimerism across full MHC barriers without graft-versus-host disease following in utero bone marrow transplantation in pigs. Exp Hematol 2005;33:371–379.
  14. Peranteau WH, Hayashi S, Hsieh M, Shaaban AF, Flake AW: High-level allogeneic chimerism achieved by prenatal tolerance induction and postnatal non-myeloablative bone marrow transplantation. Blood 2002;100:2225–2234.
  15. Crombleholme TM, Harrison MR, Zanjani ED: In utero transplantation of hematopoietic stem cells in sheep: the role of T cells in engraftment and graft-versus-host disease. J Pediatr Surg 1990;25:885–892.
  16. Pixley JS, Zanjani ED, Shaft DM, Porada C, Mackintosh FR: Prolonged hematopoietic chimerism in normal mice transplanted in utero with human hematopoietic stem cells. Pathobiology 1998;66:230–239.
  17. Carlson BM: Fetal period and birth; in Human Embryology and Development. New York, Churchill Livingstone, 1999, pp 447–468.
  18. Maddox JF, Mackay CR, Brandon MR: Ontogeny of ovine lymphocytes. I. An immunohistological study on the development of T lymphocytes in the sheep embryo and fetal thymus. Immunology 1987;62:97–105.
  19. Owen RD: Immunogenetic consequences of vascular anastomoses between bovine twins. Science 1945;102:400–401.
  20. Cancro MP: Crossing the threshold of lymphocyte activation. J Immunol 2005;174:5159–5160.
  21. Schwartz RH, Mueller DL: Immunological tolerance; in Paul WE (ed): Fundamental Immunology. New York, Lippincott-Raven Press, 1999, pp 901–963.
  22. Hermiston ML, Xu Z, Weiss A: CD45: a critical regulator of signaling thresholds in immune cells. Annu Rev Immunol 2003;21:107–137.
  23. Maddox JF, Mackay CR, Brandon MR: The sheep analogue of leucocyte common antigen. Immunology 1985;55:347–353.
  24. Flake AW, Zanjani ED: In utero hematopoietic stem cell transplantation: ontogenic opportunities and biologic barriers. Blood 1999;94:2179–2191.
  25. Laufer TM, Glimcher LH, Lo D: Using thymus anatomy to dissect T-cell repertoire selection. Semin Immunol 1999;11:65–70.
  26. Naquet P, Naspetti M, Boyd R: Development, organization and function of the thymic medulla in normal, immunodeficient or autoimmune mice. Semin Immunol 1999;11:47–55.
  27. Trop S, Charron J, Arguin C, Lesage S, Hugo P: Thymic selection generates T cells expressing self-reactive TCRs in the absence of CD45. J Immunol 2000;165:3073–3079.
  28. Nemazee DA, Burki K: Clonal deletion of B lymphocytes in a transgenic mouse bearing anti-MHC class I antibody genes. Nature 1989;337:562–566.
  29. Rodriguez-Pinto D: B cells as antigen presenting cells. Cell Immunol 2005;238:67–75.
  30. Roux E, Dumont-Girard F, Starobinski M, Siegrist CA, Helg C, Chapuis B, Roosnek E: Recovery of immune reactivity after T-cell-depleted bone marrow transplantation depends on thymic activity. Blood 2000;96:2299–2303.
  31. De Kleer I, Vastert B, Klein M, Teklenburg G, Arkesteijn G, Yung GP, Albani S, Kuis W, Wulffraat N, Prakken B: Autologous stem cell transplantation for autoimmunity induces immunologic self-tolerance by reprogramming autoreactive T cells and restoring the CD4+CD25+ immune regulatory network. Blood 2006;107:1696–1702.
  32. Wardemann H, Nussenzweig MC: B-cell self-tolerance in humans. Adv Immunol 2007;95:83–110.
  33. Singh NJ, Schwartz RH: Primer: mechanisms of immunologic tolerance. Nat Clin Pract Rheumatol 2006;2:44–52.
  34. Carrier E, Lee TH, Busch MP, Cowan MJ: Induction of tolerance in nondefective mice after in utero transplantation of major histocompatibility complex-mismatched fetal hematopoietic stem cells. Blood 1995;86:4681–4690.
  35. Kim HB, Shaaban AF, Milner R, Fichter C, Flake AW: In utero bone marrow transplantation induces donor-specific tolerance by a combination of clonal deletion and clonal anergy. J Pediatr Surg 1999;34:726–729.
  36. Sefrioui H, Donahue J, Srivastava AS, Gilpin E, Lee TH, Carrier E: Alloreactivity following in utero transplantation of cytokine-stimulated hematopoietic stem cells: the role of recipient CD4– cells. Exp Hematol 2002;30:617–624.
  37. Fairchild PJ, Waldmann H: Extrathymic signals regulate the onset of T-cell repertoire selection. Eur J Immunol 2000;30:1948–1956.
  38. Larsen WJ: Development of the head and neck; in Human Embryology. Cincinnati, Churchill Livingstone, 1997, pp 347–374.
  39. Stites DP, Pavia CS: Ontogeny of human T cells. Pediatrics 1979;64:795–802.