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This Article Full Text Full Text (PDF) All Versions of this Article: 297/3/L538    most recent 00162.2009v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Solodushko, V. Articles by Fouty, B. PubMed PubMed Citation Articles by Solodushko, V. Articles by Fouty, B.

INNOVATIVE METHODOLOGY

Dexamethasone and mifepristone increase retroviral infectivity through different mechanisms

¹Center for Lung Biology and Departments of ²Pharmacology, ³Biochemistry, and ⁴Internal Medicine, University of South Alabama School of Medicine, Mobile, Alabama

Submitted 15 May 2009 ; accepted in final form 23 June 2009

ABSTRACT

Using adapted retroviruses for gene delivery is a modern and powerful tool in biological research as well as a promising approach for gene therapy. An important limitation for the extensive use of retroviral vectors is the low infection rate in target cells such as pulmonary vascular endothelial cells due to the insufficient infectivity of standard retrovirus supernatants that can only be overcome by complicated methods of virus concentration. This paper describes two easy methods to augment target cell infectivity, first by increasing the retroviral titer in the medium collected from packaging cells by stimulation of viral propagation with dexamethasone, and second, by increasing target cell sensitivity to retroviral infection by the glucocorticoid receptor antagonist, mifepristone. Using this method, we increased the infectivity of pulmonary microvascular endothelial cells from 16% to 85%. We demonstrate that mifepristone increased the susceptibility of target cells to retroviruses without increasing the viral titer of the supernatant. Dexamethasone, but not mifepristone, increased expression of delivered proteins such as GFP that are important for early identification of infected cells. Each, or both step(s), may be included in a standard protocol for retrovirus propagation and infection of target cells.

retroviruses; pulmonary artery endothelium; gene delivery