title: Long-term cardiac gene expression using a coxsackieviral vector.
abstract: Efficient myocardial gene transfer in the intact adult heart is difficult using conventional transfer vectors. Since coxsackievirus B3 (CVB3) is cardiotropic, it may be possible to exploit its cardiotropic characteristics to design a vector for gene transfer to the intact heart. We generated a recombinant CVB3 cDNA by inserting a green fluorescent protein (GFP) gene immediately upstream from the VP0 capsid protein of CVB3. The infectious virus (rCVB3-GFP) was recovered from the supernatants of the transfected Cos-7 cells, and was grown in HeLa cells to titers of 10(11) pfu/ml. In the rCVB3-GFP infected HeLa cells and neonatal rat cardiac myocytes, GFP protein expression was documented by immunoblot and by fluorescent microscopy. GFP expression was maintained after five passages in HeLa cells. To test in vivo expression of GFP, we infected 8-week-old inbred female Balb/C mice with 10(6) pfu of rCVB3-GFP, intraperitoneally. GFP was present in up to 30% of cardiac myocytes over the 8 weeks post infection (p.i.) and it was co-localized with CVB3 infection. Surprisingly, in spite of detection of GFP up to at least 8 weeks after infection, there was no mortality in the mice. It is possible to express exogenous proteins in the intact heart after an intraperitoneal (i.p.) injection of recombinant coxsackievirus. The duration of expression persisted for at least 8 weeks with little immune response nor mortality. These results demonstrated that the cardiac tropism of CVB3 could be used to design vectors for efficient gene expression in the intact heart.
authors.affiliation: Department of Medicine, Sungkyunkwan University School of Medicine, Cardiac and Vascular Center, Samsung Medical Center, 50 Il-won Dong, Kangnam-Ku, Seoul 135-710, South Korea.
authors.forename: Byung-Kwan
authors.initials: BK
authors.lastname: Lim
authors.forename: Jae-Ok
authors.initials: JO
authors.lastname: Shin
authors.forename: Sang-Chol
authors.initials: SC
authors.lastname: Lee
authors.forename: Duk-Kyung
authors.initials: DK
authors.lastname: Kim
authors.forename: Dong-Ju
authors.initials: DJ
authors.lastname: Choi
authors.forename: Seong-Choon
authors.initials: SC
authors.lastname: Choe
authors.forename: Kirk U
authors.initials: KU
authors.lastname: Knowlton
authors.forename: Eun-Seok
authors.initials: ES
authors.lastname: Jeon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000235:genetics; D029822:Enterovirus B, Human / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009206:Myocardium / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D004274:DNA, Recombinant; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins