FIG. 5.

Distribution of tripeptidyl-peptidase I (TPP-I) activity in the CNS of nonhuman primates after CNS administration of AAVrh.10hCLN2. African green monkeys were injected with AAVrh.10hCLN2 at a dose of 1.8×10¹² genome copies in a total of 180 μl divided equally among 12 loci through 6 burr holes, or were injected with PBS in the same volume and at the same location. After sacrifice the nonhuman primate brains were hemisected. The left hemispheres were sectioned into 1-cm³ sections (referred to as cubes); these cubes were homogenized and assayed for TPP-I activity. The activity was measured as fluorescence units per minute per milligram of protein. TPP-I activity from all the cubes in the PBS control animal was averaged and this was taken to be the endogenous background. In (A–C), the lighter cubes represent regions that have an activity of TPP-I less than 2 SD above the background, and the darker cubes represent the regions that have an activity greater than 2 SD above endogenous background levels; the data are shown as a three-dimensional representation of distribution of TPP-I activity. (A) The data from the control PBS-injected animal. (B) Data from one representative animal administered AAVrh.10hCLN2 and killed at 7 days. (C) Data from one representative animal treated with AAVrh.10hCLN2 and killed at 90 days. (D) Percentage of CNS with TPP-I activity greater than 2 SD above endogenous background is shown as a bar graph for all groups in the study (n=1 PBS killed at 7 days, n=4 AAVrh.10hCLN2 killed at 7 days, n=AAVrh.10hCLN2 killed at 90 days) and is compared with the historical data obtained from our study using AAV2hCLN2 in the same setting.