1. Volume 13, Issue 5, Pages 967-975 (May 2006)
Long-term doxycycline-regulated transgene expression in the retina of nonhuman primates following subretinal injection of recombinant AAV vectors 
Knut Stieger, Guylène Le Meur, Françoise Lasne, Michel Weber, Jack-Yves Deschamps, Delphine Nivard, Alexandra Mendes-Madeira, Nathalie Provost, Laurent Martin, Philippe Moullier, Fabienne Rolling 
Molecular Therapy 
Volume 13, Issue 5, Pages (May 2006)
DOI: /j.ymthe
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

2. FIG. 1
Structure of rAAV vectors. Vectors encode the macaque erythropoietin cDNA (mEpo) under the control of the doxycycline-inducible TetO.CMV promoter and the rtTA chimeric transactivator (rtTA-M2) under the control of either the CAG promoter or the human RPE65 promoter. pA (rtTA-M2), bovine growth hormone polyadenylation signal; pA (mEpo), SV40 polyadenylation signal; WPRE, woodchuck hepatitis virus posttranscriptional regulatory element; ITR, inverted terminal repeat of AAV2. Cassettes were incorporated into AAV4 or AAV5 capsids as indicated.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

3. FIG. 2
Time course of EPO concentration in the anterior chamber fluid or vitreous of three nonhuman primates after subretinal delivery of AAV2/5.CAG.TetOn.epo. Three-day induction cycles are indicated as “Dox.” For macaque P5.1, EPO concentrations in the anterior chamber fluid and vitreous of the injected eye were compared (indicated by a circle). For macaque 5.2, squares indicate time points for which IEF patterns of EPO were evaluated (see Figs. 5A and 5B). For macaques P5.1 and P5.2: (♦) anterior chamber fluid right eye (injected), (▪) vitreous right eye (injected), (▴) anterior chamber fluid left eye (uninjected control).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

4. FIG. 3
Time course of EPO concentration in the anterior chamber fluid of four nonhuman primates after subretinal delivery of AAV2/4.CAG.TetOn.epo and AAV2/4.RPE65.TetOn.epo vectors. Three-day induction cycles are indicated as “Dox.” A square indicates the time point at which IEF patterns of EPO were evaluated (see Fig. 5C). (A) AAV2/4.CAG.TetOn.epo subretinally injected macaques P4.1 and P4.2. (B) AAV2/4.RPE65.TetOn.epo subretinally injected macaque P4.3 and P4.4.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

5. FIG. 4
Assessment of retinal morphology and function following long-term Tet-regulated EPO expression and evaluation of humoral immune response against the rtTA-M2 and EPO proteins. (A) Fundus photography and OCT image of the right eye of macaque 5.2 (injected). The green line indicates the scanning path. The white dotted circle indicates the zone of the bleb. (B) Fundus photography and OCT image of the left eye of macaque 5.2 (uninjected control). The green line indicates the scanning area of the retina. (C) Standard ERG recordings of both eyes of macaque 5.2. Graphs correspond to rod ERG, maximum ERG, cone ERG, and flicker (from the top to the bottom line). (D) Evaluation of antibody generation against rtTA-M2 and EPO by Western blot analysis. a.c.f., anterior chamber fluid; mpi, months postinjection.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

6. FIG. 5
Isoelectric focusing patterns and corresponding integrated profiles of EPO. Fluorescence intensity is reported in mega linear arbitrary units (MLAU). (A, B) AAV2/5.CAG.TetOn.epo subretinally injected macaque P5.1. (C) AAV2/4.CAG.TetOn.epo subretinally injected macaque P4.1. (D) AAV2/1.CAG.TetOn.epo intramuscularly injected primate (Mac13 in [23]). (E) Uninjected macaque.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions