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Gene delivery to in situ veins: Differential effects of adenovirus and adeno-associated viral vectors  Mohammad H. Eslami, MD, Sidhu P. Gangadharan, MD,

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Presentation on theme: "Gene delivery to in situ veins: Differential effects of adenovirus and adeno-associated viral vectors  Mohammad H. Eslami, MD, Sidhu P. Gangadharan, MD,"— Presentation transcript:

1 Gene delivery to in situ veins: Differential effects of adenovirus and adeno-associated viral vectors  Mohammad H. Eslami, MD, Sidhu P. Gangadharan, MD, XinXin Sui, MD, Kurt K. Rhynhart, MD, Richard O. Snyder, PhD, Michael S. Conte, MD  Journal of Vascular Surgery  Volume 31, Issue 6, Pages (June 2000) DOI: /mva Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

2 Fig. 1 Schematic representation of transgene constructs used. Adenovirus (Ad ) constructs were inserted into the E1 region of an E1, E3-deleted adenoviral backbone (36 kB). Adeno-associated virus (AAV ) constructs are inserted between the AAV inverted terminal repeats, replacing the AAV coding sequences (4.7 kB). Journal of Vascular Surgery  , DOI: ( /mva ) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

3 Fig. 2 Analysis of rabbit jugular veins after exposure to adenovirus (A, B, C) or adeno-associated virus (D, E, F) vectors. Specimens were explanted 2 days (A, B) and 14 days (C) after exposure to adenovirus vectors (1·109/mL; β-galactosidase [A, C ]; green fluorescent protein [B] ). A, Inset is a representative area on cross section, demonstrating localization of blue staining to the endothelial monolayer (magnification, 1000×). B, En-face fluorescent microscopy of an adenovirus-green fluorescent protein infected vein is shown. Representative areas of adeno-associated virus-β-galactosidase (1·109/mL) infected veins are shown on days 2 (D) , 14 (E) , and 39 (F) postexposure. F, Inset (magnification, 400×; L, lumen) is a cross section from a densely stained area, demonstrating positive-staining endothelial and smooth muscle cells at day 39. (Magnifications: A and C, 7.5×; B, 200×; D, 20×; E and F, 49×). Journal of Vascular Surgery  , DOI: ( /mva ) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

4 Fig. 3 Reverse transcriptase-polymerase chain reaction analysis of the green fluorescent protein transgene in target tissue. A, Representative of adeno-associated virus-infected specimens (day 2 , N = 3; day 7 , N = 1; day 14, N = 2; day 30, N = 3). B, Adenovirus specimens (day 2, N = 3; day 7, N = 2; day 14, N = 2; day 30, N = 2). RNA was isolated from veins on the indicated days after viral exposure. Polymerase chain reaction amplification was performed, by using both reverse transcribed (+) and non-reverse transcribed (– ) templates (paired lanes for each tissue sample). M, Size markers (50 bp); P, green fluorescent protein-containing plasmid control; V, control (uninfected) jugular vein; T, testicle from an experimental animal. Journal of Vascular Surgery  , DOI: ( /mva ) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

5 Fig. 4 Results of ex vivo THP-1 adhesion assays, expressed as a ratio to sham-infected vein segments on the indicated days after vector exposure. Adhesion to adeno- virus-infected veins on postexposure day 2 was significantly increased (P <.05), as compared with adeno-associated virus-treated (AAV ) veins and adenovirus infection at all other time points. POD, Postexposure day. Journal of Vascular Surgery  , DOI: ( /mva ) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

6 Fig. 5 Histologic and immunohistochemical analysis of explanted veins after adenovirus or adeno-associated virus mediated gene transfer. A, B, C, Vein specimen harvested 2 days after adeno-associated virus-green fluorescent protein exposure. A, CD31 stain; B, ICAM-1 stain; and C, RAM-11 stain (all magnifications, 100×). D, An adeno-associated virus-green fluorescent protein exposed vein on day 7, ICAM-1 stain (magnification, 100×; inset magnification, 1000×). E (adeno-associated virus-green fluorescent protein) and F (adenovirus-green fluorescent protein), Representative histologic sections from a 30-day explanted vein, stained only with hematoxylin (magnification, 40×). Journal of Vascular Surgery  , DOI: ( /mva ) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

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