Volume 57, Issue 5, Pages (May 2000)

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Volume 57, Issue 5, Pages 1973-1980 (May 2000) Gene transfer targeting interstitial fibroblasts by the artificial viral envelope-type hemagglutinating virus of Japan liposome method Michiko Tsujie, Yoshitaka Isaka, Yutaka Ando, Yoshitaka Akagi, Yasufumi Kaneda, Naohiko Ueda, Enyu Imai, Masatsugu Hori Kidney International Volume 57, Issue 5, Pages 1973-1980 (May 2000) DOI: 10.1046/j.1523-1755.2000.00047.x Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 1 Representative fluorescence photomicrographs of FITC-labeled oligodeoxynucleotide (ODN)-transfected kidneys. FITC-labeled phosphorothioate ODNs accumulated in the nuclei of interstitial cells 10 minutes after retrograde transfection via the ureter by the artificial viral envelope (AVE)-type hemagglutinating virus of Japan (HVJ) liposome method (A). To examine the cellular localization of the transfected ODNs, rabbit polyclonal antitype IV collagen antibodies and rhodamine-conjugated donkey antirabbit IgG were used to stain the basement membrane (B). FITC (green)-positive nuclei were seen outside of the basement membrane (red). To identify the transduced cells, the monoclonal antibody ER-TR7 (C) and ED-1 (D) were used. FITC-labeled ODNs were seen in the nuclei of the fibroblasts (C), but not coincident with macrophages (D).◂ Kidney International 2000 57, 1973-1980DOI: (10.1046/j.1523-1755.2000.00047.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 2 Delivery into interstitial area. The representative appearances of kidneys show that 100 μL of infusion (A) resulted in the medulla-limited India-ink distribution; however, 300 μL of infusion (B) induced diffuse distribution in the cortex. India ink was scattered in the interstitial area, but not in the tubular lumen (C). Kidney International 2000 57, 1973-1980DOI: (10.1046/j.1523-1755.2000.00047.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 3 β-galactosidase activity in interstitial cells. HVJ-liposomes containing thepEBAct-NlacF gene were introduced into the kidney retrogradely via the ureter. Seven (A) and 14 (B) days after transfection, nuclear localized blue stain was dominantly observed in the interstitial cells by X-gal staining (arrowheads). Kidney International 2000 57, 1973-1980DOI: (10.1046/j.1523-1755.2000.00047.x) Copyright © 2000 International Society of Nephrology Terms and Conditions