Virus Research Institute and Dana Farber Cancer Institute seeks support for pre-clinical gene transfer studies utilizing peripheral mononuclear cells, purified CD4+ T-cells from uninfected and HIV-1 infected individuals that will result in """"""""Intracellular immunization"""""""" against HIV-1. The research proposal will optimize large scale retroviral gene transfer of a intracellular single-chain antibody against the HIV-1 envelope, required for the proceeding into a clinical setting. The vector LNCs105 carries the selection marker neoR and the human single-chain antibody F105. Vectors and packaging cell lines have been produced. Optimal conditions for gene transfer into primary cells will be established in cells from HIV-1 infected and non-infected individuals. Testing HIV-1 replication in long term cultures to evaluate the ability to suppress HIV-1 replication in cells from HIV-1 patients and large clinical scale transduction of CD4+ T lymphocytes will be done. Gene transfer, expression, cell selection and expansion in """"""""dry runs"""""""" of the protocol and the level of cell-surface receptor expression, morphology, cytokine profile, and response to antigen stimulation will be evaluated. DFCI """"""""gene transfer facility"""""""" and personnel will participate in this pre- clinical arm and in future clinical proposals.
