Gene transfer targeting interstitial fibroblasts by the artificial viral envelope-type hemagglutinating virus of Japan liposome method.
BACKGROUND
Tubulointerstitial inflammation and fibrosis are commonly associated with most human glomerular diseases. The degree of tubulointerstitial damage, rather than the glomerular injury, could correlate with the degree of renal functional impairment and accurately predict long-term prognosis. In an effort to understand the pathogenesis of the progressive interstitial fibrosis, we developed a new strategy of gene transfer to the interstitial fibroblasts.


METHODS
Either fluorescein isothiocyanate (FITC)-labeled oligodeoxynucleotides (ODNs) or pEBAct-NlacF expression vector was introduced into the kidney of normal rats retrogradely via ureter by using the artificial viral envelope (AVE)-type hemagglutinating virus of Japan (HVJ) liposome method.


RESULTS
FITC-labeled ODNs were accumulated diffusely in the nuclei of the interstitial cells in the transfected kidney 10 minutes after transfection, and the interstitial cells were identified as interstitial fibroblasts by immunostaining with ER-TR7. To examine the gene expression in the interstitium, pEBAct-NlacF gene-conjugated HVJ liposome was injected retrogradely through the ureter, and in consequence, nuclear beta-galactosidase activity was continuously observed in interstitial cells at least two weeks after transfection.


CONCLUSIONS
This new strategy of gene transfer to the interstitial fibroblasts is useful for the investigation of the pathophysiology of tubulointerstitial lesion, and furthermore, it may be a promising new therapeutic method for the progression of interstitial fibrosis.
Tsujie
M
M
Department of Internal Medicine and Therapeutics, and Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.
Isaka
Y
Y
Ando
Y
Y
Akagi
Y
Y
Kaneda
Y
Y
Ueda
N
N
Imai
E
E
Hori
M
M
D000818:Animals; D005347:Fibroblasts / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D018014:Gene Transfer Techniques; D004854:Herpesvirus 4, Human / Q000235:genetics; D007668:Kidney / Q000378:metabolism*; D008081:Liposomes / Q000008:administration & dosage; D008297:Male; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012093:Replicon; D018108:Respirovirus / Q000235:genetics*
D008081:Liposomes; D009838:Oligodeoxyribonucleotides; D016650:Fluorescein-5-isothiocyanate