Gene transfer of Fc-fusion cytokine by in vivo electroporation: application to gene therapy for viral myocarditis.
Among a number of techniques for gene transfer in vivo, the direct injection of plasmid DNA into muscle is simple, inexpensive and safe. Although combining direct DNA injection with in vivo electroporation increases the efficiency of gene transfer into muscle, applications of this method have remained limited because of the relatively low expression level. To overcome this problem, we developed a plasmid vector that expresses a secretory protein as a fusion protein with the noncytolytic immunoglobulin Fc portion and used it for electroporation-mediated viral interleukin 10 (vIL-10) expression in vivo. The fusion cytokine vIL-10/mutFc was successfully expressed and the peak serum concentration of vIL-10 was almost 100-fold (195 ng/ml) higher than with a non-fusion vIL-10 expression plasmid. The expressed fusion cytokine suppressed the phytohemagglutinin-induced IFN-gamma production by human peripheral blood mononuclear cells and decreased the mortality in a mouse viral myocarditis model as effectively as vIL-10 expression. These results demonstrate that the transfer of plasmid DNA expressing a noncytolytic Fc-fusion cytokine is useful to deliver enhanced levels of cytokine without altering general biological activities. This simple and efficient system should provide a new approach to gene therapy for human diseases and prove very useful for investigating the function of newly discovered secretory protein genes.
Adachi
O
O
Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Nakano
A
A
Sato
O
O
Kawamoto
S
S
Tahara
H
H
Toyoda
N
N
Yamato
E
E
Matsumori
A
A
Tabayashi
K
K
Miyazaki
J
J
D000818:Animals; D018274:Electroporation / Q000379:methods*; D004686:Encephalomyocarditis virus; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D016753:Interleukin-10 / Q000097:blood / Q000235:genetics*; D008297:Male; D051379:Mice; D008811:Mice, Inbred DBA; D023421:Models, Animal; D009205:Myocarditis / Q000276:immunology / Q000628:therapy* / Q000821:virology; D010957:Plasmids / Q000008:administration & dosage*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage
D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D016753:Interleukin-10