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Prevention of the second stage of epithelial loss is a potential novel treatment for bronchiolitis obliterans  Yunge Zhao, MD, PhD, John F. Steidle, BA,

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Presentation on theme: "Prevention of the second stage of epithelial loss is a potential novel treatment for bronchiolitis obliterans  Yunge Zhao, MD, PhD, John F. Steidle, BA,"— Presentation transcript:

1 Prevention of the second stage of epithelial loss is a potential novel treatment for bronchiolitis obliterans  Yunge Zhao, MD, PhD, John F. Steidle, BA, Gilbert R. Upchurch, MD, Irving L. Kron, MD, Christine L. Lau, MD  The Journal of Thoracic and Cardiovascular Surgery  Volume 145, Issue 4, Pages e1 (April 2013) DOI: /j.jtcvs Copyright © Terms and Conditions

2 Figure 1 Representative pictures of hematoxylin and eosin staining of the allograft from 3, 7, 12, and 21 days after transplantation. The magnification of all the pictures was 40×. Black arrows indicate epithelial cells. The gray arrows indicate basal cells. The donors are Balb/c mice and the recipients are Rag-1 KO and C57BL/6 wild-type mice. C57BL/6 to C57BL/6 isograft controls are also performed. The Journal of Thoracic and Cardiovascular Surgery  , e1DOI: ( /j.jtcvs ) Copyright © Terms and Conditions

3 Figure 2 Losing of epithelial cells in the transplanted trachea. Epithelial cells are disappearing rapidly owing to ischemia–reperfusion injury in the early stage (days 1-3 after tracheal transplantation) in all transplanted trachea. The pseudoepithelial cells are regenerated from days 4 to 7 in all allografts. The second loss of the regenerated epithelium occurred from days 7 to 14 in all of the allografts except in Rag-1 KO mice, which show a pattern similar to that of isograft controls. The Journal of Thoracic and Cardiovascular Surgery  , e1DOI: ( /j.jtcvs ) Copyright © Terms and Conditions

4 Figure 3 Immunohistochemical staining of GFP in allografts on day 7 after tracheal transplantation. A, Representative picture of GFP staining in allograft (Balb/c to GFP/C57BL/6) on day 7. B, Representative picture of GFP staining in allograft (GFP/C57BL/6 to Balb/c) on day 7. Cells stained brown/dark brown indicate GFP-positive cells. All sections were counterstained lightly with hematoxylin for viewing negatively stained cells. Normal IgG was used as controls. Black arrows indicate epithelial cells. The grey arrows indicate basal cells. The magnifications are indicated in the pictures. GFP, Green fluorescent protein; IgG, immunoglobulin G. The Journal of Thoracic and Cardiovascular Surgery  , e1DOI: ( /j.jtcvs ) Copyright © Terms and Conditions

5 Figure 4 Immunohistochemical staining of CD3+ T-cells in (A) the allografts of Balb/c to GFP/C57BL/6 and in (B) isograft of C57BL/6 to C57BL/6. Cells stained brown indicate CD3+ T-cell infiltration. All sections were counterstained lightly with hematoxylin for viewing negatively stained cells. The slides were stained with anti-CD3 antibody. Normal IgG was used as control. The magnifications are indicated in the pictures. GFP, Green fluorescent protein; IgG, immunoglobulin G; BC, Balb/c. The Journal of Thoracic and Cardiovascular Surgery  , e1DOI: ( /j.jtcvs ) Copyright © Terms and Conditions

6 Figure 5 A, Injection of recipient epithelial cells prevents the second epithelial loss on day 12 (top panel) when compared with noninjected mice (bottom panel). B, Comparison of luminal obliteration of the allografts with and without epithelial injection on day 21. Data shown are the mean ± standard deviation. *P = .007, n = 6. BC, Balb/c; EPC, epithelial cells. The Journal of Thoracic and Cardiovascular Surgery  , e1DOI: ( /j.jtcvs ) Copyright © Terms and Conditions

7 Figure E1 Histologic examination of epithelial cells after digestion with 0.01% collagenase A in the trachea of GFP/C57BL/6 mice. The epithelial layers are intact in the nondigested normal trachea and 15-minutes digestion trachea. Epithelial cells detached after digestion of 30 minutes and completely detached after 1 hour of digestion. Black arrows indicate epithelial cells. The gray arrows indicate basal cells. The magnifications are indicated on the pictures. GFP, Green fluorescent protein. The Journal of Thoracic and Cardiovascular Surgery  , e1DOI: ( /j.jtcvs ) Copyright © Terms and Conditions

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