Article Figures & Data
Figures
- Fig. 1.
Construction of the hexon chimerasdlAd5NCAT-H2 and dlAd5NCAT-H2L2 by recombination. (A) Schematic and amino acid alignment of the Ad5 and Ad2 hexon proteins showing the locations of conserved regions (open bars) and hypervariable regions (HVR; gray bars) and the numbers of amino acid residues (aa) per region. Identity is shown as percent with the number of identical residues shown in parentheses. (B) Virus construction.dlAd5NCAT DNA was digested with either Bsu36I orDrdI to generate left- and right-hand-end subgenomic fragments, respectively. The fragments were cotransfected with linearized pH2-5. Shaded boxes, Ad5 hexon sequence; black boxes, regions of homology between the plasmid and the subgenomic fragments; thin lines, plasmid sequence. (C) Plasmid constructs for introducing mutations into the Ad5 hexon gene. Plasmid pH5 contains theHindIII-KpnI fragment (map units [m.u.] 51.03 to 61.81) of Ad5.
- Fig. 2.
Transduction of A549 lung cells by Ad hexon chimeras. (A) Schematics of the hexon proteins encoded by dlAd5NCAT,dlAd5NCAT-H2, and dlAd5NCAT-H2L2. Open bars, conserved sequence; black bars, Ad5 unique sequence; crosshatched bars, Ad2 unique sequence. (B) CAT activity in extracts of A549 cells infected with 10 or 100 particles of dlAd5NCAT,dlAd5NCAT-H2, or dlAd5NCAT-H2L2 per cell. CAT activity is expressed as the calculated total activity in the total extract. Values are means ± standard deviations of triplicates.
- Fig. 3.
Effect of immunization with Ad hexon or fiber chimeras on the in vivo transduction efficiency of dlAd5NCAT. (A) Evaluation of the efficacy of intravenous administration. Adult female Sprague-Dawley rats were administered 1012 particles ofdlAd5NCAT-H2, dlAd5NCAT-H2L2, ordlAd5NCAT-F7 via the jugular vein, and the livers were assayed for CAT activity 3 days later. Data are the means of triplicates ± standard deviations. (B) Readministration. Adult female Sprague-Dawley rats were administered 1012particles of dlAd5NCAT-H2, dlAd5NCAT-H2L2, ordlAd5NCAT-F7 via the jugular vein. At 28 days postinfection, the same animals received 1012 particles ofdlAd5NCAT via the jugular vein and were sacrificed 24 h later, and their livers were processed for CAT activity assays. Values shown are a compilation of two independent experiments and are the means ± 1 standard deviation. Abbreviations: F7,dlAd5NCAT-F7; H2, dlAd5NCAT-H2; H2L2,dlAd5NCAT-H2L2.
