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Laboratory and Animal Investigations |

Pleural Space as a Site of Ectopic Gene Delivery*: Transfection of Pleural Mesothelial Cells With Systemic Distribution of Gene Product

Clinton J. Devin, BS; Y.C. Gary Lee, MBChB; Richard W. Light, MD, FCCP; Kirk B. Lane, PhD
Author and Funding Information

*From the Pulmonary and Critical Care Division (Mr. Devin and Dr. Lane), Vanderbilt University, Nashville, TN; and the Department of Pulmonary Medicine (Drs. Light and Lee), St. Thomas Hospital, Nashville, TN.

Correspondence to: Richard W. Light, MD, FCCP, Department of Pulmonary Medicine, St. Thomas Hospital, 4220 Harding Rd, Nashville, TN 37205; e-mail: rlight98@yahoo.com



Chest. 2003;123(1):202-208. doi:10.1378/chest.123.1.202
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Study objectives: Successful ectopic gene therapy requires the transfection of the cells at the ectopic site, with local and systemic delivery of the gene product. This study aimed to evaluate the pleural mesothelial surface as a potential site for ectopic gene therapy.

Design: A secreted placental alkaline phosphatase (PALP) plasmid was injected bilaterally into the pleural spaces of seven rabbits via a chest tube, while an irrelevant reporter plasmid was injected into seven control rabbits. Blood was collected at baseline and at 24, 48, and 72 h after the injections. Pleural fluid was collected by lavage at 24, 48, and 72 h after the injections. The PALP level was measured by chemiluminesence.

Measurements and results: Significant expressions of PALP proteins were observed in the serum of the treatment rabbits, with a threefold increase over baseline at 24 h, a ninefold increase at 48 h, and a twofold increase at 72 h. The serum PALP levels in the control rabbits remained at baseline levels at all time points. The pleural fluid PALP levels peaked at 24 h and decreased over the next 72 h. Mimicking the in vivo pattern, pleural mesothelial cells transfected in vitro demonstrated a similar increase in PALP levels.

Conclusions: The results of the present short-term pilot study suggest that pleural mesothelial cells can be successfully transfected with plasmids, with increases in both the local and systemic levels of the gene product. The pleural space should be further evaluated for ectopic gene therapy.

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