LIPIDATED ANTIBODIES FOR INTRACELLULAR THERAPY OF HIV

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Effective HIV-1 therapy is a complex problem which may require at least two different approaches. We propose to address the issues of blocking viral replication in conjunction with initiation of immunoreconstitution. A novel technology has been developed which covalently links a lipoamine, glycyldioctadecylamide, to the carbohydrate moieties of polyclonal or monoclonal antibodies. The carbohydrate residues are located on the fc portion of the antibody, removed from the hypervariable region, and consequently do not interfere with antibody binding. The presence of the lipoamines appear to promote passage of the antibody through the plasma membrane of living cells without loss of epitope specificity. This has been demonstrated visually by direct staining of intracellular cytoskeletal proteins. These antibodies also contain the capability of neutralizing intracellular protein function. Preliminary data indicates that individually, lipidated anti-Tat, anti-p24/25 gag, and anti-reverse transcriptase antibodies can markedly reduced viral replication of HIV- 1IllB and wild-type isolates in infected SupT1 cells and latently infected J1.1 and OM 10.1 cells. In addition, we have cloned a lymphocyte competence growth factor specific for CD4+ cells. This lymphokine (LCF) induces the expression of IL-2R and renders the cells responsive to IL-2 stimulation. The focus of the first two aims is to optimize in vitro conditions for lipidated antibody blocking of HIV-l replication, and optimize the CD4 + lymphocyte growth potential of LCF. This includes screening a panel of lipidated anti-HIV-1 viral protein antibodies for their ability to block HIV-1 replication, individually or in different combinations. Based on the in vitro data, our third aim is to apply these two novel approaches directed towards the evaluation of antibody blocking and immunoreconstitution in a SCID mouse animal model of HIV-1.