![Page 1: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/1.jpg)
Cord Blood-Derived Regulatory T Cells:A Novel Cure for Graft-Versus-Host-
Disease
Siddharth Damania2007
![Page 2: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/2.jpg)
University of MinnesotaCancer Center
![Page 3: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/3.jpg)
Bone-Marrow and Stem-Cell Transplants
http://www.vacationusarentals.com/images/usa_map.gif
6,000 Americans
![Page 4: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/4.jpg)
Bone-Marrow and Stem-Cell Transplants
http://www.southalabama.edu/nursing/MSN%20Info/surgery_nurse.jpg
![Page 5: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/5.jpg)
Bone-Marrow and Stem-Cell Transplants
http://www.southalabama.edu/nursing/MSN%20Info/surgery_nurse.jpg
Over half develop GVHD
![Page 6: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/6.jpg)
Graft-Versus-Host-Disease (GVHD)
• GVHD occurs when transplanted cells reject host cells in the patient
Patient CellTransplanted
Cell
![Page 7: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/7.jpg)
GVHD
GVHD
GVHD
GVHD
GVHD
Purpose
• To use T-reg cells to cure mice with human GVHD
![Page 8: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/8.jpg)
First Goal
• To devise a new protocol to derive and purify CD4+25+ T-reg cells using human umbilical cord blood
![Page 9: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/9.jpg)
Types of T cells
• Cytotoxic T cells• Helper T cells• T-reg cells
http://www.britannica.com/eb/art/print?id=85110&articleTypeId=0
![Page 10: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/10.jpg)
CD4+25+ T-reg Cells
• Sakaguchi (2005)– Increased capability to suppress
immune responses– Low numbers increase risk of
autoimmune diseases
![Page 11: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/11.jpg)
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
![Page 12: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/12.jpg)
Umbilical Cord Blood
• Godfrey et al (2005)– T-reg cells isolated from peripheral
blood are often impure and require multiple purification processes
![Page 13: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/13.jpg)
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
![Page 14: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/14.jpg)
Procedure
![Page 15: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/15.jpg)
Procedure
![Page 16: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/16.jpg)
Procedure
![Page 17: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/17.jpg)
Procedure
![Page 18: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/18.jpg)
Procedure
CD4+25+ T-reg Cells
![Page 19: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/19.jpg)
3/28 KT32
KT32 -41 BBL KT32 -OX40
FACS T-reg Cell Purity (CD4/CD25)
86.2% 95.4%
91.5% 81.0%
![Page 20: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/20.jpg)
Second Goal
• To culture derived CD4+25+ T-reg cells
![Page 21: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/21.jpg)
Second Goal
• To culture derived CD4+25+ T-reg cells
CD4+25+ T-reg Cells
![Page 22: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/22.jpg)
Second Goal
• To culture derived CD4+25+ T-reg cells
CD4+25+ T-reg Cells
![Page 23: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/23.jpg)
Second Goal
• To culture derived CD4+25+ T-reg cells
CD4+25+ T-reg Cells
CD4+25+ T-reg Cells
![Page 24: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/24.jpg)
3/28 Cell Line
![Page 25: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/25.jpg)
KT32 Cell Line
![Page 26: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/26.jpg)
KT32-41BBL Cell Line
![Page 27: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/27.jpg)
KT32-OX40L Cell Line
![Page 28: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/28.jpg)
Culture Expansion of T-reg Cells
KT-32-OX40:y =
504.24e0.7776x
R2 = 0.7983
3/28:y =
10715e0.5056x
R2 = 0.8031
KT32-41BBL:y =
6.4462x5.0346
R2 = 0.8129
KT32:y =
8077.2x1.5986
R2 = 0.40660.00E+00
1.00E+06
2.00E+06
3.00E+06
4.00E+06
5.00E+06
6.00E+06
0 2 4 6 8 10 12 14
Day (After Umbilical Cord Blood Derivation)
Number of Cells
3/28'KT32KT32-OX40KT32-41BBLExpon. (KT32-OX40)Expon. (3/28')Power (KT32-41BBL)Power (KT32)
![Page 29: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/29.jpg)
Culture Decline of T-reg Cells
3/28:
y = 3E+16x-8.5021
R2 = 0.6171
KT32:y = -14345x + 304762
R2 = 0.9924
KT32-OX40:y = -58036x + 2E+06
R2 = 0.3654
KT32-41BBL:y = -30548x +
719881
R2 = 0.5407
0.00E+00
1.00E+06
2.00E+06
3.00E+06
4.00E+06
5.00E+06
6.00E+06
14 15 16 17 18 19 20 21 22
Days (After Umbilical Cord Blood Derivation)
Number of Cells
3/28'
KT32
KT32-OX40
KT32-41BBL
Power (3/28')
Linear (KT32)
Linear (KT32-OX40)Linear (KT32-41BBL)
![Page 30: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/30.jpg)
Third Goal
• To analyze the histological effect of CD4+25+ T-reg cells in GVHD mice
![Page 31: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/31.jpg)
Procedure
![Page 32: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/32.jpg)
Procedure
![Page 33: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/33.jpg)
Procedure
GVHD
GVHD
GVHD
GVHD
GVHD
![Page 34: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/34.jpg)
Procedure
GVHD
GVHD
GVHD
GVHD
GVHD
![Page 35: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/35.jpg)
Procedure
GVHD
GVHD
GVHD
GVHD
GVHD
![Page 36: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/36.jpg)
Healthy Liver
![Page 37: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/37.jpg)
Diseased Liver
![Page 38: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/38.jpg)
Diseased Liver
![Page 39: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/39.jpg)
Diseased Liver
![Page 40: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/40.jpg)
Diseased Liver
![Page 41: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/41.jpg)
Diseased Liver
![Page 42: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/42.jpg)
GVHD Reduction with T-reg Cells
Liver
GVHD Lung
GVHD Spleen GVHD
Colon GVHD
GVHD Mice 3.8 3.4 3.0 2.0 T-reg Treated Mice 1.9 1.9 3.6 1.1
![Page 43: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/43.jpg)
GVHD Reduction with T-reg Cells
Liver
GVHD Lung
GVHD Spleen GVHD
Colon GVHD
GVHD Mice 3.8 3.4 3.0 2.0 T-reg Treated Mice 1.9 1.9 3.6 1.1
![Page 44: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/44.jpg)
GVHD Reduction with T-reg Cells
Liver
GVHD Lung
GVHD Spleen GVHD
Colon GVHD
GVHD Mice 3.8 3.4 3.0 2.0 T-reg Treated Mice 1.9 1.9 3.6 1.1
![Page 45: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/45.jpg)
GVHD Reduction with T-reg Cells
Liver
GVHD Lung
GVHD Spleen GVHD
Colon GVHD
GVHD Mice 3.8 3.4 3.0 2.0 T-reg Treated Mice 1.9 1.9 3.6 1.1
50%
![Page 46: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/46.jpg)
GVHD Reduction with T-reg Cells
Liver
GVHD Lung
GVHD Spleen GVHD
Colon GVHD
GVHD Mice 3.8 3.4 3.0 2.0 T-reg Treated Mice 1.9 1.9 3.6 1.1
44%
![Page 47: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/47.jpg)
GVHD Reduction with T-reg Cells
Liver
GVHD Lung
GVHD Spleen GVHD
Colon GVHD
GVHD Mice 3.8 3.4 3.0 2.0 T-reg Treated Mice 1.9 1.9 3.6 1.1
20%
![Page 48: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/48.jpg)
GVHD Reduction with T-reg Cells
Liver
GVHD Lung
GVHD Spleen GVHD
Colon GVHD
GVHD Mice 3.8 3.4 3.0 2.0 T-reg Treated Mice 1.9 1.9 3.6 1.1
45%
![Page 49: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/49.jpg)
Future Work
• Optimize culture expansion
![Page 50: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/50.jpg)
Future Work
• Optimize culture expansion • Increase rate of cure in mice
![Page 51: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/51.jpg)
Future Work
• Optimize culture expansion • Increase rate of cure in mice• Test T-reg cells as a therapeutic
cure for GVHD in humans
![Page 52: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/52.jpg)
Acknowledgements• Dr. Keli Hippen• Dr. Paul Harker-Murray• The University of Minnesota• Lois Fruen• Advanced Science Research
QuickTime™ and aTIFF (Uncompressed) decompressor
are needed to see this picture.
QuickTime™ and aTIFF (Uncompressed) decompressor
are needed to see this picture.QuickTime™ and a
TIFF (Uncompressed) decompressorare needed to see this picture.
![Page 53: Cord Blood-Derived Regulatory T Cells: A Novel Cure for Graft-Versus-Host-Disease](https://reader036.vdocument.in/reader036/viewer/2022062410/56815465550346895dc27f0c/html5/thumbnails/53.jpg)
Cord Blood-Derived Regulatory T Cells:A Novel Cure for Graft-Versus-Host-
Disease
Siddharth Damania2007