Modulation of adenovirus-mediated gene transfer by nitric oxide

American Journal of Respiratory Cell and Molecular Biology
I Y HaddadS Matalon

Abstract

We assessed the role of .NO in recombinant adenovirus-mediated gene transfer both in vitro and in vivo. NIH3T3 fibroblasts, stably transfected with the human inducible nitric oxide synthase, but lacking tetrahydrobiopterin (NIH3T3/iNOS [inducibile nitric oxide synthase]), were infected with replication-deficient adenovirus (E1-deleted), containing either the luciferase or the Lac Z reporter genes (AdCMV-Luc and AdCMV-Lac Z; 1-10 plaque forming units [pfu]/cell). Incubation of infected cells with sepiapterin (50 microM), a precursor of tetrahydrobiopterin, progressively increased nitrate/nitrite levels in the medium and decreased both luciferase and beta-galactosidase protein expression to approximately 60% of their corresponding control values, 24 h later. NIH3T3/iNOS cells had normal ATP (adenosine 5'-triphosphate) levels and did not release LDH(lactic dehydrogenase) into the medium. Pretreatment of these cells with N(G)-monomethyl-L-arginine (L-NMMA; 1 mM), an inhibitor of iNOS, prevented the sepiapterin-mediated induction of .NO and restored gene transfer to baseline values. Incubation of NIH3T3/iNOS with 8-bromo-cGMP (400 microM) in the absence of sepiapterin, or exposure of AdCMV-Luc to large concentrations of .NO, did not...Continue Reading

Citations

Dec 19, 2000·Free Radical Biology & Medicine·D R Janero, J F Ewing
May 1, 2007·Clinics in Chest Medicine·Viranuj SueblinvongDaniel J Weiss
Sep 21, 2004·Respiratory Research·Anne-Karin HesseFritz Krombach
Nov 14, 1997·The American Journal of Physiology·S M ManuelS Matalon
Jul 17, 1999·The American Journal of Physiology·T JillingS Matalon

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