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J Thorac Cardiovasc Surg 2003;126:1155-1161
© 2003 The American Association for Thoracic Surgery

General thoracic surgery

Adenovirus encoding soluble tumor necrosis factor {alpha} receptor immunoglobulin prolongs gene expression of a cotransfected reporter gene in rat lung

Takashi Suda, MDa, Tsutomu Tagawa, MDa, Samer A. Kanaan, MDa, Benjamin D. Kozower, MDa, Niccolò Daddi, MDa, T. Mohanakumar, PhDb, G. Alexander Patterson, MD, FRCS(C)a,*

a Division of Cardiothoracic Surgery, Washington University School of Medicine, Barnes Jewish Hospital, St Louis, Mo, USA
b Department of Surgery, Washington University School of Medicine, Barnes Jewish Hospital, St Louis, Mo, USA

Received for publication October 25, 2002; revisions received December 27, 2002; revisions received May 20, 2003; accepted for publication June 3, 2003.

* Address for reprints: G. Alexander Patterson, MD, Division of Cardiothoracic Surgery, Washington University School of Medicine, One Barnes-Jewish Hospital Plaza, 3108 Queeny Tower, St. Louis, MO 63110, USA
pattersona{at}msnotes.wustl.edu

OBJECTIVE: Because almost all pulmonary diseases are not caused by one gene, multiple gene transfection is required for current gene therapy. Adenovirus is an important gene therapy vector, but a short duration and the inability of repeated administration remain limitations. The aims of this study were to evaluate whether adenoviral vector encoding soluble tumor necrosis factor {alpha} receptor immunoglobulin and ß-galactosidase cotransfection prolongs gene expression and facilitates repeated vector administration to investigate the feasibility of a cotransfection strategy.

METHODS: F344 rats received intratracheal administration of 1 x 109 plaque-forming units of adenoviral vector encoding ß-galactosidase or both adenoviral vector encoding ß-galactosidase and adenoviral vector encoding soluble tumor necrosis factor {alpha} receptor immunoglobulin. In the expression study ß-galactosidase gene expression in the lung was examined by means of enzyme-linked immunosorbent assay on days 2, 7, 14, 28, and 56 (n = 4/day). In the repeated transfection study, soluble tumor necrosis factor {alpha} receptor immunoglobulin and ß-galactosidase were readministered once (7 days after the first adenovirus administration) or twice (on days 7 and 14; n = 4/day). A 2-way factorial analysis of variance was used for statistical analysis.

RESULTS: Soluble tumor necrosis factor {alpha} receptor immunoglobulin and ß-galactosidase cotransfection prolonged the duration of ß-galactosidase expression. However, antiadenovirus antibody production was significantly increased in the cotransfection group. In addition, there was no increase in ß-galactosidase expression after readministration of soluble tumor necrosis factor {alpha} receptor immunoglobulin and ß-galactosidase.

CONCLUSION: Adenoviral vector encoding soluble tumor necrosis factor {alpha} receptor immunoglobulin and ß-galactosidase cotransfection prolongs ß-galactosidase expression but does not increase ß-galactosidase expression after repeated administration. These results suggest that tumor necrosis factor {alpha} is one of the most important factors in regulating the duration of gene expression. The cotransfection approach is feasible, but the increase of antiadenovirus antibodies might make repeated cotransfection unfeasible.




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