Peritransplant Ischemic Injury Is Associated With Up-Regulation of Stromal Cell-Derived Factor-1
Mohamad H. Yamani, MD*,*,
Norman B. Ratliff, MD ,
Daniel J. Cook, PhD ,
E. Murat Tuzcu, MD*,
Yang Yu, MS*,
Robert Hobbs, MD*,
Gustavo Rincon, MD*,
Corinne Bott-Silverman, MD*,
James B. Young, MD*,
Nicholas Smedira, MD and
Randall C. Starling, MD, MPH*
* Department of Cardiovascular Medicine, Kaufman Center for Heart Failure, Cleveland Clinic Foundation, Cleveland, Ohio.
Department of Anatomic Pathology, Kaufman Center for Heart Failure, Cleveland Clinic Foundation, Cleveland, Ohio.
Department of Cardiothoracic Surgery, Kaufman Center for Heart Failure, Cleveland Clinic Foundation, Cleveland, Ohio.
Department of Allogen Laboratory, Kaufman Center for Heart Failure, Cleveland Clinic Foundation, Cleveland, Ohio.
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Figure 1 Fluorescent in situ hybridization and double immunofluorescence of initial heart biopsy specimen from ischemia group showing DAPI blue-stained nuclei containing Y chromosome (arrows) in green (A). (B) These cells are positive for c-kit in red (arrowheads), suggestive of stem cell origin.
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Figure 2 A heart biopsy specimen from the control group, obtained at one year after transplantation, showing immunostaining for CD45RB (A) and CD68 (B) in red. Y chromosome (arrows) is detected in green.
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Figure 3 A heart biopsy specimen from the ischemia group, obtained at one year after transplantation, showing immunostaining for myosin heavy chain. Y chromosome is detected (arrow) in a cardiomyocyte nucleus.
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Figure 4 Heart biopsy specimens from the ischemia group, obtained at one year after transplantation, showing immunostaining for smooth muscle alpha-actin (A) and factor VIII (B). Y chromosomes (arrows) are detected in vascular smooth muscle cell nuclei in intramyocardial arteriole (A) and capillary endothelial cell (B).
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Figure 5 Stromal cell-derived factor-1 immunohistochemistry of initial heart biopsy specimens from the control (A) and ischemia (B) groups.
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Figure 6 Stromal cell-derived factor-1 (SDF-1) immunoblotting showing increased myocardial protein expression (normalized to glyceraldehyde phospho dehydrogenase [GAPDH]) in the presence of ischemic injury (I) compared to the control group (C). Liver cirrhotic tissue was used as a positive control.
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Figure 7 TaqMan polymerase chain reaction (PCR) showing messenger ribonucleic acid (mRNA) expression (left) of stromal cell-derived factor-1 (SDF-1) in the control (upper panel) and ischemia (lower panel) groups with the corresponding ribosomal ribonucleic acid (rRNA) (right) as internal control. The ischemia group demonstrates a shift of the mRNA curve to the left, indicating an earlier threshold of detection on the PCR thermal cycle and thus an increased mRNA expression of SDF-1. Rn = change in ribonucleic acid acitivity.
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Figure 8 Kaplan-Meier curves of freedom from vasculopathy (A) and survival (B) in the control (solid line) and ischemia (dashed line) groups.
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