Adenovirus-Mediated Transgene Expression in Nonhuman Primate Brain
Bohn, MC and Choi-Lundberg, DL and Davidson, BL and Leranth, C and Kozlowski, DA and Smith, JC and O'Banion, MK and Redmond, DE, Adenovirus-Mediated Transgene Expression in Nonhuman Primate Brain, Human Gene Therapy, 10, (7) pp. 1175-1184. ISSN 1043-0342 (1999) [Refereed Article]
Transgene expression in the brain of St. Kitts green monkey, Cercopithecus aethiops sabeus, was studied following injection of a serotype 5 adenoviral vector deleted in E1 and E3. The vector harbored the transgene for Escherichia coli β-galactosidase (β-Gal) with the simian virus 40 (SV40) nuclear localization signal under control of the Rous sarcoma viral (RSV) long terminal repeat. Several titers ranging from 5 x 107 to 2 x 109 plaque-forming units (PFU) in volumes ranging from 5 to 250 μl were injected into the caudate nuclei of 18 monkeys. Monkeys were treated with dexamethasone for 9 days, beginning the day prior to surgery, and were sacrificed at 1 week or at 1, 2, or 3 months. At 1 week, β-Gal was expressed in thousands of cells, including both neurons and astrocytes. In addition, some dopaminergic neurons in the substantia nigra expressed transgene, suggesting retrograde transport of the vector. At 1 month 162,000 ± 68,000 (SEM) or 65,000 ± 29,000 β-Gal-expressing cells persisted in striatum injected with 6 x 108 PFU in 30 μl or 5 x 107 PFU in 5 μl, respectively. Transgene expression was also observed in one of two monkeys sacrificed at 2 months and in a single monkey sacrificed at 3 months. No transgene expression was observed at 1 month in striatum injected with a higher titer (2 x 109 PFU in 100 μl) or more dilute vector (5 x 107 PFU in 30 μl). Staining for the major histocompatibility complex II (MHC II) subtype DR showed intense staining in sites injected with a higher vector titer, in which no transgene persisted at 1 month, whereas low to moderate staining was present in sites with high transgene expression. These observations suggest that there is an optimal range of vector titers for obtaining persistent transgene expression from E1E3-deleted adenovirus in primate brain, above which host responses limit transgene stability.