1. Volume 116, Issue 3, Pages 658-665 (March 1999)
Protective and therapeutic effect of DNA-based immunization against hepadnavirus large envelope protein 
Christine Rollier, Claire Sunyach, Luc Barraud, Nora Madani, Catherine Jamard, Christian Trepo, Lucyna Cova 
Gastroenterology 
Volume 116, Issue 3, Pages (March 1999)
DOI: /S (99)
Copyright © 1999 American Gastroenterological Association Terms and Conditions

2. Fig. 1
DNA vaccination vector and DHBV large protein expression. (A) Schematic representation of the pCI-preS/S plasmid encoding the DHBV large protein. (B) Expression of the DHBV large protein from pCI-preS/S plasmid in transiently transfected LMH cells. Proteins from cell lysates were probed in immunoblotting analysis with rabbit anti-DHBpreS antibody. Arrow indicates the 36-kilodalton DHBV large protein. Lane 1, concentrated serum-derived DHBV particles; lane 2, mock (LMH cells); lane 3, pCMV-DHBV–transfected LMH cells; lane 4, pCI-transfected LMH cells; lane 5, pCI-preS/S–transfected LMH cells.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

3. Fig. 1
DNA vaccination vector and DHBV large protein expression. (A) Schematic representation of the pCI-preS/S plasmid encoding the DHBV large protein. (B) Expression of the DHBV large protein from pCI-preS/S plasmid in transiently transfected LMH cells. Proteins from cell lysates were probed in immunoblotting analysis with rabbit anti-DHBpreS antibody. Arrow indicates the 36-kilodalton DHBV large protein. Lane 1, concentrated serum-derived DHBV particles; lane 2, mock (LMH cells); lane 3, pCMV-DHBV–transfected LMH cells; lane 4, pCI-transfected LMH cells; lane 5, pCI-preS/S–transfected LMH cells.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

4. Fig. 2
Kinetics of humoral response in ducks after immunization with (A) pCI-preS/S plasmid: 100 μg (ducks 301 and 302), 200 μg (duck 303), and 400 μg (ducks 305 and 306; duck 305 accidentally died at week 5). (B) DHBpreS polypeptide (100 μg) emulsified in Freund's adjuvant. Detection of anti-preS end point titer antibodies, expressed as log10, was performed using a direct ELISA test. Graphs show data from individual responders ducks. The first injection was administered at time 0; filled arrows represent DNA or protein boosts for all ducks. The open arrow represents the delayed DNA boost of 100 μg administered to ducks 301 and 302.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

5. Fig. 3
Recognition in immunoblotting assay of the recombinant DHBpreS protein, used in the ELISA test, by (A) serum from pCI-preS/S–immunized duck and (B) serum from duck immunized with concentrated DHBV particles. Arrowheads show the p21 DHBpreS protein, p33 DHBpreS precursor, and its p28 degradation product.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

6. Fig. 4
Recognition in immunoblotting test of DHBV particles. VP lanes, concentrated serum-derived viral particles; Mock lanes, concentrated uninfected duck serum. The lanes were shown with (A) serum from pCI-preS/S–immunized duck, (B) polyclonal anti-DHBpreS rabbit antiserum, and (C) control duck serum from pCI-immunized duck. Arrowhead indicates the 36-kilodalton DHBV large protein.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

7. Fig. 5
Recognition in immunoblotting assay of proteins from DHBV-infected and -uninfected duck livers shown with (A) serum from pCI-preS/S–immunized duck, (B) polyclonal anti-DHBpreS rabbit antiserum, and (C) control duck serum from pCI-immunized duck. Arrowheads indicate the p36 DHBV large protein, its p28 cleavage product, and the migration location of p18 DHBV small protein, based on its detection by anti-S monoclonal antibody in this immunoblotting assay (not shown).
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

8. Fig. 6
In vitro neutralization of DHBV infectivity with immunized duck sera. Primary duck hepatocytes were infected with DHBV inoculum preincubated either with phosphate-buffered saline (– –), PCI-immunized control duck serum ( ), or pCI-preS/S–immunized duck serum (—). The release of viral particles was quantified in supernatants of cell culture by dot-blot hybridization. Means of duplicate determinations are presented. Samples of day 8 of phosphate-buffered saline control were not available.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

9. Fig. 7
In vivo protection of ducklings from DHBV infection. Southern blot analysis of total liver DNA (20 μg/lane) extracted from ducklings inoculated with DHBV preincubated with the following: (A) control duck serum (lanes 1 and 2); (B) 100 μg pCI-preS/S–immunized duck serum (lanes 3–6); (C) 200 μg pCI-preS/S–immunized duck serum (lanes 7–10); and (D) 400 μg pCI-preS/S–immunized duck serum (lanes 11–14). Each lane represents liver DNA of 1 inoculated duckling. Lane 15, control DHBV DNA linearized by EcoRI. L, linear; RC, relaxed circular; SS, single-stranded DHBV DNA forms.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

10. Fig. 8
Effect of DNA immunization on duck viremia. Viremia was followed up for 42 weeks in 6 pCI-preS/S–immunized, 3 unimmunized, and 3 pCI-immunized DHBV-carrier ducks. The level of mean serum DHBV DNA for each group of ducks, based on individual viremia titers, is represented. Arrows show DNA injections (100 μg/duck) at weeks 4, 7, 11, and 28.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

11. Fig. 9
Effect of DNA immunization on intrahepatic DHBV DNA. Southern blot analysis of liver DNA (20 μg/lane) extracted from (A) 5 control ducks and (B) 6 pCI-preS/S–immunized ducks. Relative band intensity for total DHBV DNA was quantified by densitometry. Lower panel shows the specific detection of DHBV cccDNA pool. L, linear; RC, relaxed circular; SS, single-stranded DHBV DNA forms.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions