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Annual and Interim Progress Report Summaries

Principal Investigator: James Wilson

Project: Impact of Adenovirus Infections on Vaccine Efficacy for HIV-1

Submitted June 1, 2012

The study comparing different AAV capsids for muscle transduction is completed. Based on the reporter expression data AAV2/8 was chosen as a preferred vector for delivery of NAbs into muscle. The team is now in the process of performing follow up analyses of the collected tissue samples with the goal to determine the relationship between transgene expression level, mRNA expression level and transgene genome copy numbers. This analysis will facilitate the development of the optimal bNAb gene constructs by allowing scientists to define the gene structure components that need most improvement.

Comparative expression study for AAV2/8 mediated transduction of rhesus NAb in three species of non-human primates (rhesus macaques, African green and squirrel monkeys) is ongoing. Samples were collected weekly and expression levels for up to 84 days post transduction for rhesus macaques, and up to 54 days post transduction for African green and squirrel monkeys were determined. Highest expression is observed in rhesus macaques, closely followed by squirrel monkeys, with African green monkeys expressing at the lowest levels. This set of data will determine which NHP species will progress to the next series of studies aimed at defining the optimal conditions for transduction of skeletal muscle (target muscle group, vector formulation and promoter).

The study on evaluation of the pre-existing NAbs to capsid on transgene expression in rhesus macaques is completed. The team determined that pre-existing NAbs with titer of up to 1/20 do not significantly affect expression of transgene in muscle, while higher titers of pre-existing NAbs (1/320) decrease peak expression by at least 50%.

14 additional configurations of NAbs were developed, 8 of which were de novo synthesized and cloned into expression vectors. The remaining NAbs are in the process of being cloned. The team is now performing comparative expression studies in RAG KO mice.

Submitted July 1, 2010
Submitted October 16, 2009 (Interim Report)
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