Incorporation of adenovirus in calcium phosphate precipitates enhances gene transfer to airway epithelia in vitro and in vivo.
Adenovirus (Ad)-mediated gene transfer to airway epithelia is inefficient because the apical membrane lacks the receptor activity to bind adenovirus fiber protein. Calcium phosphate (CaPi) precipitates have been used to deliver plasmid DNA to cultured cell lines. However, such precipitates are not effective in many primary cultures or in vivo. Here we show that incorporating recombinant adenovirus into a CaPi coprecipitate markedly enhances transgene expression in cells that are resistant to adenovirus infection. Enhancement requires that the virus be contained in the precipitate and viral proteins are required to increase expression. Ad: CaPi coprecipitates increase gene transfer by increasing fiber-independent binding of virus to cells. With differentiated cystic fibrosis (CF) airway epithelia in vitro, a 20-min application of Ad:CaPi coprecipitates that encode CF transmembrane conductance regulator produced as much CF transmembrane conductance regulator Cl- current as a 24-h application of adenovirus alone. We found that Ad:CaPi coprecipitates also increased transgene expression in mouse lung in vivo; importantly, expression was particularly prominent in airway epithelia. These results suggest a new mechanism for gene transfer that may be applicable to a number of different gene transfer applications and could be of value in gene transfer to CF airway epithelia in vivo.
Fasbender
A
A
Howard Hughes Medical Institute and Departments of Internal Medicine and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
Lee
J H
JH
Walters
R W
RW
Moninger
T O
TO
Zabner
J
J
Welsh
M J
MJ
D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D019556:COS Cells; D002130:Calcium Phosphates / Q000008:administration & dosage*; D002213:Capsid / Q000502:physiology; D036022:Capsid Proteins; D011232:Chemical Precipitation; D003550:Cystic Fibrosis / Q000235:genetics / Q000628:therapy*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008168:Lung / Q000821:virology*; D051379:Mice; D008810:Mice, Inbred C57BL
D002130:Calcium Phosphates; D036022:Capsid Proteins; C485828:alpha-tricalcium phosphate; C018315:hexon capsid protein, Adenovirus; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; C485829:calcium phosphate, dibasic, anhydrous