1. Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease Siddharth Damania 2007

2. University of Minnesota Cancer Center

3. Bone-Marrow and Stem-Cell Transplants http://www.vacationusarentals.com/images/usa_map.gif 6,000 Americans

4. Bone-Marrow and Stem-Cell Transplants http://www.southalabama.edu/nursing/MSN%20Info/surgery_nurse.jpg

5. Bone-Marrow and Stem-Cell Transplants http://www.southalabama.edu/nursing/MSN%20Info/surgery_nurse.jpg Over half develop GVHD

6. Graft-Versus-Host-Disease (GVHD) GVHD occurs when transplanted cells reject host cells in the patient Patient Cell Transplanted Cell

7. GVHD Purpose To use T-reg cells to cure mice with human GVHD

8. First Goal To devise a new protocol to derive and purify CD4 + 25 + T-reg cells using human umbilical cord blood

9. Types of T cells Cytotoxic T cells Helper T cells T-reg cells http://www.britannica.com/eb/art/print?id=85110&articleTypeId=0

10. CD4 + 25 + T-reg Cells Sakaguchi (2005) –Increased capability to suppress immune responses –Low numbers increase risk of autoimmune diseases

11. CD4 + 25 + T-reg Cells Sakaguchi (2005) –Increased capability to suppress immune responses –Low numbers increase risk of autoimmune diseases Suri-Payer et al (2006) –Potential to suppress multiple types of autoimmune diseases

12. Umbilical Cord Blood Godfrey et al (2005) –T-reg cells isolated from peripheral blood are often impure and require multiple purification processes

13. Umbilical Cord Blood Godfrey et al (2005) –T-reg cells isolated from peripheral blood are often impure and require multiple purification processes –Umbilical cord blood facilitates easier purification and presents purer quantities of T-reg cells

14. Procedure

18. CD4 + 25 + T-reg Cells

19. FACS T-reg Cell Purity (CD4/CD25) 86.2%95.4% 91.5%81.0%

20. Second Goal To culture derived CD4 + 25 + T-reg cells

21. Second Goal To culture derived CD4 + 25 + T-reg cells CD4 + 25 + T-reg Cells

22. Second Goal To culture derived CD4 + 25 + T-reg cells CD4 + 25 + T-reg Cells

23. Second Goal To culture derived CD4 + 25 + T-reg cells CD4 + 25 + T-reg Cells

24. 3/28 Cell Line

25. KT32 Cell Line

26. KT32-41BBL Cell Line

27. KT32-OX40L Cell Line

28. Culture Expansion of T-reg Cells

29. Culture Decline of T-reg Cells

30. Third Goal To analyze the histological effect of CD4 + 25 + T-reg cells in GVHD mice

31. Procedure

33. GVHD

34. Procedure GVHD

35. Procedure GVHD

36. Healthy Liver

37. Diseased Liver

42. GVHD Reduction with T-reg Cells

45. 50%

46. GVHD Reduction with T-reg Cells 44%

47. GVHD Reduction with T-reg Cells 20%

48. GVHD Reduction with T-reg Cells 45%

49. Future Work Optimize culture expansion

50. Future Work Optimize culture expansion Increase rate of cure in mice

51. Future Work Optimize culture expansion Increase rate of cure in mice Test T-reg cells as a therapeutic cure for GVHD in humans

52. Acknowledgements Dr. Keli Hippen Dr. Paul Harker-Murray The University of Minnesota Lois Fruen Advanced Science Research

53. Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease Siddharth Damania 2007