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ministered via tail vein injection immediately after clamp release and on postoperative days 2 and 4. Another group of rats underwent sham thoracotomies without vessel clamping. On postoperative day 5, animals were sacrificed and lung tissue specimens were analyzed with light and fluorescence microscopy for the presence of MSC-eGFP and SDF-1. Quantitative analysis of lung tissue and serum for SDF-1 was achieved with RT-PCR and ELISA techniques, respectively. MSC were examined for the presence of CXCR4, the receptor to SDF-1, using immunocytochemistry. Results: IRI lung tissue demonstrated greater levels of MSC-eGFP as examined by immunohistologic analysis with both light and fluorescence microscopy as compared with uninjured lung tissue. There was a 2.5-fold increase in the quantity of SDF-1 in the IRI lungs compared to the uninjured lungs (p 0.01) as determined by quantitative RT-PCR. Examination of serum SDF-1 levels using ELISA showed no difference between animals subjected to lung IRI and sham thoracotomy. However, lung tissue analysis for SDF-1 by quantitative RT-PCR demonstrated significantly higher levels in IRI lungs compared to control lungs. The presence of CXCR4 on MSC was confirmed with immunocytology. Conclusions: Circulating MSC can populate IRI lungs. SDF-1 is upregulated locally in IRI lungs and may serve to recruit CXCR4 expressing MSC to the injured lung tissue.

The recently published manuscript by Nussbaum et al. (1) describes teratoma formation and the generation of a host immune response following transplantation of undifferentiated murine embryonic stem cells into the heart. These are important findings, emphasizing the hurdles that need to be overcome before embryonic stem cell therapy can become a clinical reality. We thought it proper to remind the readership of The FASEB Journal of our study detailing undifferentiated embryonic stem cell transplantation into the heart published in 2005 (2). In this study, we presented evidence that undifferentiated embryonic stem cells transplanted into the heart both form teratomas and elicit an immune response by the host. We specifically classified the immune response by examining cellular infiltrates into the heart and presented proof of teratoma formation in both syngeneic and allogeneic recipients. We were somewhat surprised, then, that Dr. Nussbaum’s manuscript does not refer to this previously published work while presenting similar conclusions to those put forth by our study.

Cardiothoracic Surgery/Falk Research Center, Stanford University Medical School, Stanford, CA, USA Department of Radiology, Stanford University Medical School, Stanford, CA, USA Geron Corporation, Menlo Park, CA, USA Division of Thoracic and Cardiovascular Surgery, Hannover Medical School, Carl Neuberg Street 1, 30625 Hannover, Germany www.elsevier.com/locate/ejcts European Journal of Cardio-thoracic Surgery 29 (2006) 1069

Purpose: Ischemia/reperfusion (IR) injury in lung transplantation is a major cause of acute graft dysfunction, and is characterized by the production of pro-inflammatory cytokines such as IL-1 , and apoptosis of donor respiratory epithelium. We tested the hypothesis that intratracheal administration of adenovirus, overexpressing the human anti-apoptotic protein Bcl-2, will inhibit IR injury to donor lung allografts. Procedures: PVG (RT1c) rats underwent in-vivo intratracheal administration of 1.0x10pfu of either AdvBcl-2 in PBS (n 7), AdvNull in PBS (n 6) or PBS alone (n 8). The left lungs were procured after 24hrs of in-vivo incubation. After 1hr (Group I) or 24hrs (Group II, AdvBcl-2 in PBS vs. AdvNull in PBS) of cold ischemia, donor lungs were orthotopically transplanted into ACI (RT1a) recipients and reperfused for 2hrs. Peak inspiratory pressures (PIPs) were measured, and grafts were excised and analyzed by ELISA, activated caspase-3 assay and histology. Donor pulmonary vein paO2 was measured in Group II. Results: Human Bcl-2 protein was overexpressed in AdvBcl-2 treated lungs compared to AdvNull and PBS alone treated lungs at the time of transplantation (p 0.05), and localized in the alveolar epithelium. The Bcl-2 overexpression in AdvBcl-2 lungs were identical in Groups I and II . In Group I, AdvBcl-2 treated lungs demonstrated reduced PIPs compared to AdvNull and PBS alone treated controls (p 0.01), decreased intragraft IL-1 cytokine (p 0.05), cytochrome c (p 0.01), caspase-3 activity (p 0.05), and ISOL TUNNEL positive nuclei (1.7% 0.7% vs. 4.7% 1.1%, and 6.1% 0.4%, p 0.05). In Group II animals, AdvBcl-2 lungs maintained significant decreases in IL-1 production, caspase-3 activity, and ISOL TUNNEL positive nuclei (3.6% 0.8% vs.19.3% 3.6%) and higher donor pulmonary vein paO2 (p 0.05) compared to AdvNull lungs. Conclusions: Intratracheal adenoviral overexpression of human Bcl-2 limits ischemia/reperfusion injury, improves post-transplant lung function and may serve as a valuable gene therapy regimen in human lung transplantation.

SPIRIT 2013 checklist: recommended items to address in a clinical trial protocol and related documents. (DOC 112Â kb)

SPIRIT 2013 checklist: recommended items to address in a clinical trial protocol and related documents. (DOC 112Â kb)