Patent ID: 8247167

Claim:
An in vitro, cell-based method of identifying compounds capable of activating latent HIV-1 comprising: Providing an isolated SUPT1 cell line comprising a latent, HIV-1-derived provirus vector comprising: An in-frame insertion of a secretable alkaline phosphatase (seap) gene open reading frame in place of the HIV-1 env gene start codon, an inactivation of the vpu gene start codon, a deletion of about 2500 base pairs of the HIV-1 pol gene, and an insertion of an egfp coding sequence 10 base pairs upstream from the nef gene start codon, wherein upon activation, the vector expresses the seap, a secretable marker for late viral gene expression, or the egfp, a marker for early viral gene expression; providing a candidate compound and a positive control compound for late viral gene activation, wherein the positive control compound is selected from tumor necrosis factor-α, phorbol 12-myristate 13-acetate or valproic acid; combining the cell line with the candidate compound and combining the cell line with the positive control compound; monitoring for seap expression in the presence of the candidate compound as compared to seap expression in the absence of the candidate compound and monitoring for seap expression in the presence of the positive control to determine if the compound is capable of activating the vector late viral expression genes; monitoring for egfp expression in the presence of the candidate compound as compared to egfp expression in the absence of the candidate compound and monitoring for egfp expression in the presence of the positive control to determine if the compound is capable of activating the vector early viral expression genes; and wherein detecting increased seap expression by monitoring an amount of seap expression that is greater in the presence of a candidate compound than in the absence of a candidate compound and/or detecting increased egfp expression by monitoring an amount of egfp expression that is greater in the presence of a candidate compound than in the absence of a candidate compound; and wherein detecting increased seap expression by monitoring an amount of seap expression that is greater in the presence of the positive control compound than in the absence of a the positive control compound and detecting increased egfp expression by monitoring an amount of egfp expression that is greater in the presence of the positive control compound than in the absence of the positive control compound, identifies a compound capable of activating latent HIV-1.