Gene delivery of l-caldesmon protects cytoskeletal cell membrane integrity against adenovirus infection independently of myosin ATPase and actin assembly.
The cytoskeleton is critical to the viral life cycle. Agents like cytochalasin inhibit viral infections but cannot be used for antiviral therapy because of their toxicity. We report the efficacy, safety, and mechanisms by which gene delivery of human wild-type low-molecular-weight caldesmon (l-CaD) protects cell membrane integrity from adenovirus infection in a DF-1 cell line, an immortalized avian fibroblast that is null for l-CaD. Transfection with an adenovirus (Ad)-controlled construct mediated a dose-dependent decline in transcellular resistance. In accordance with a computational model of cytoskeletal membrane properties, Ad disturbed cell-cell and cell-matrix adhesion and membrane capacitance. Transfection with the Ad-l-CaD construct attenuated adenovirus-mediated loss in transcellular resistance. Quantitation of vinculin-stained plaques revealed an increase in total focal contact mass in monolayers transfected with the Ad-l-CaD construct. Expression of l-CaD protected transcellular resistance through primary effects on membrane capacitance and independently of actin solubility and effects on pre-stress, as measured by the decline in isometric tension in response to cytochalasin D. Expression of l-CaD exhibited less Trypan blue cell toxicity than cytochalasin, and, unlike cytochalasin, it did not interfere with wound closure or adversely effect transcellular resistance. These findings demonstrate the gene delivery of wild-type human l-CaD as a potentially efficacious and safe agent that inhibits some of the cytopathic effects of adenovirus.
Haxhinasto
Kari
K
Department of Internal Medicine, C33 GH, University of Iowa College of Medicine, Iowa City, IA 52242, USA.
Kamath
Anant
A
Blackwell
Ken
K
Bodmer
James
J
Van Heukelom
Jon
J
English
Anthony
A
Bai
Er-Wei
EW
Moy
Alan B
AB
D000199:Actins / Q000378:metabolism; D000256:Adenoviridae / Q000472:pathogenicity; D000257:Adenoviridae Infections / Q000473:pathology* / Q000628:therapy; D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology* / Q000821:virology; D003599:Cytoskeleton / Q000502:physiology* / Q000821:virology; D025802:Electric Capacitance; D005347:Fibroblasts / Q000378:metabolism / Q000821:virology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D008954:Models, Biological; D009218:Myosins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D000199:Actins; D002148:Calmodulin-Binding Proteins; D009218:Myosins