1. Volume 18, Issue 1, Pages 126-134 (January 2010)
The Pleiotropic Effects of Natural AAV Infections on Liver-directed Gene Transfer in Macaques 
Lili Wang, Roberto Calcedo, Huan Wang, Peter Bell, Rebecca Grant, Luk H Vandenberghe, Julio Sanmiguel, Hiroki Morizono, Mark L Batshaw, James M Wilson 
Molecular Therapy 
Volume 18, Issue 1, Pages (January 2010)
DOI: /mt
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

2. Figure 1
GFP expression in rhesus macaque livers after intravenous infusion of 3 × 1012 GC/kg of AAV.TBG.EGFP pseudotyped with AAV8, hu.37, and rh.8 capsids. (a) Livers are harvested at day 7. (b) Livers are harvested at day 35. Representative images of each animal are shown. Bar = 100 µm.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

3. Figure 2
In vitro neutralizing antibody assay on preinjected monkey sera. Serial twofold dilutions of monkey and naive mouse serum were incubated with 1 × 109 genome copies of (a) AAVrh.8.CMV.LacZ or (b) AAV8.CMV.LacZ. Transduction efficiency at each serum dilution was measured by β-galactosidase levels (relative light unit per second, RLU/second) using a luminometer 24 hours after infection. AAV neutralization titer for each sample was determined by the highest serum dilution that inhibited AAV.CMV.LacZ transduction by ≥50%, compared with the mouse serum control. Pre-samples for , , and were obtained 14 days before gene transfer, and 12 days before gene transfer for and AAV, adeno-associated virus.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

4. Figure 3
Detection of neutralizing AAV antibodies in preinjected monkey sera by in vivo passive transfer experiment. C57BL/6 mice were infused with of 200 µl of serum sample from an individual NHP or naive mouse serum. Two hours after the passive transfer, mice received an intravenous injection of 1 × 109 genome copies of AAV.LSP.cFIX-W vector packaged with the respective capsids (AAV8, hu.37, or rh.8). cFIX expression levels in the plasma at days 7, 21, and 28 after vector administration are shown. Data are presented as mean ± SD (n = 3). AAV, adeno-associated virus; cFIX, canine factor IX; NHP, nonhuman primate.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

5. Figure 4
Time course of liver enzyme levels (ALT and AST) in monkeys before and after intravenous infusion of 3 × 1012 genome copies/kg of AAV.TBG.EGFP pseudotyped with AAV8, hu.37, and rh.8. Pre1 samples were taken 71, 79, and 87 days before vector administration of AAV8, hu.37, and rh.8, respectively. Pre2 samples were taken 12, 20, and 35 days before vector administration of AAV8, hu.37, and rh.8, respectively. AAV, adeno-associated virus; ALT, alanine aminotransferase; AST, aspartate aminotransferase; LFT, liver function test.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

6. Figure 5
Time course of T-cell responses to AAV capsid and GFP in the PBMCs of monkeys after vector administration. PBMCs, isolated before and weekly after vector injection, were stimulated with AAV8 capsid and GFP-peptide libraries and assayed by IFNγ ELISPOT. Background from unstimulated control (<30 SFU/106 lymphocytes) was subtracted from each sample. Asterisk indicates a positive response, which is defined as more than threefold of background and >55 SFU/106 lymphocytes. AAV, adeno-associated virus; ELISPOT, enzyme-linked immunosorbent assay; GFP, green fluorescent protein; IFNγ, interferon-γ; PBMC, peripheral blood mononuclear cell; SFU, spot forming units.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

7. Figure 6
T-cell response to GFP in different tissues at day 35 after vector administration. PBMCs and lymphocytes isolated from spleen, bone marrow (BM), and liver were stimulated with GFP-peptide library and assayed by IFNγ ELISPOT. Background from unstimulated control (<30 SFU/106 PBMCs, <65 SFU/106 splenocytes, <70 SFU/106 bone marrow lymphocytes, and <50 SFU/106 liver lymphocytes) was subtracted from each sample. Asterisk indicates a positive response, which is defined as more than threefold of background and >55 SFU/106 lymphocytes. AAV, adeno-associated virus; ELISPOT, enzyme-linked immunosorbent assay; GFP, green fluorescent protein; IFNγ, interferon-γ; PBMC, peripheral blood mononuclear cell; SFU, spot forming units.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions