This SPIRAT core proposal is designed to further research that has resulted from a collaboration between Apollon and the Institute for Biotechnology and Advanced Medicine at the University of Pennsylvania. These efforts recently yielded reports describing the successful immunization of mice and rabbits against the human immunodeficiency virus type I (HIV-1) coat protein (gp160) using the technique of facilitated genetic inoculation. Animals were directly inoculated with an expression vector encoding the HIV-1gp 160 gene, which induced in humoral and cellular immunity to the HIV-1 gp 160 in vivo. More recent work has demonstrated protection from a normally lethal cell challenge with a murine myeloma expressing gp 160, in addition, the same expression vector injected in non-human primates has been shown to cause seroconversion, antigen specific T cell proliferation, and specific CTL generation against the env encoded proteins.
The aim of the research described is to extend these studies to the rhesus macaque and chimp animal models and to the treatment of HIV-1 infection of humans. For the present proposal, various plasmids will be constructed that will express a variety of different SIV or HIV-1 encoded proteins. Plasmids will be modeled after the one used i the successful gp160 HIV experiments; however, the genes for several of the SIV and HIV proteins will be incorporated into the collection plasmids in order to elicit a broadly reactive immune response. The HIV constructs will be very similar to those constructed for SIV encoded protein expression, however, the HIV constructs will be modified to be conceptually safe for human inoculation in the proposed clinical trials. The immediate objective of this Molecular Biology Core proposal will be (i) construction and structural analysis of the plasmids; (ii) evaluation of the levels of protein expression in human rhabdomyosarcoma and COS-7 cells that have been transfected with the construct of interest; (iii) the production of large scale amounts of the product plasmids for animals and clinical studies according to cGMPs; (iv) support of the evaluation of clinical samples by monitoring viral load prior to and following the administration of the GeneVax plasmids. The long-term goal of this work will be the development and commercialization GENEVAX-HIV for the possible treatment of HIV-infected persons.
