HIV-1 latency model for high throughput screening
Isolated, latently infected T cell lines are provided that can be utilized in high throughput screening to discover compounds capable of activating HIV-I. The T cell lines harbor a latent HIV-I derived vector pro virus, which upon activation expresses a marker for late viral gene expression due to the insertion of the marker gene in the position of HIV-I envelope.
1. An isolated, latently infected T cell line harboring a latent HIV-I derived vector pro virus, which upon activation of the pro virus expresses a secretable marker for late viral gene expression, the gene for said marker being inserted in the position of HIV-I envelope.
2. The cell line of claim 1 , wherein the marker is a secretable enzyme.
3. The cell line of claim 2 , wherein the secretable enzyme is alkaline phosphatase.
4. The cell line of claim 3 , wherein the secreted alkaline phosphatase is capable of being detected using chemiluminescence.
5. The cell line of claim 1 , wherein the pro virus further includes a gene marker for viral early gene expression at the single cell level.
6. The cell line of claim 5 , wherein the expressed marker for viral early gene expression is a fluorescent protein.
7. The cell line of claim 6 , wherein the expressed marker for viral early gene expression is enhanced green fluorescent protein (egfp).
8. The cell line of claim 1 , wherein the provirus is capable of being activated by stimuli selected from the group consisting of tumor necrosis factor (TNF)-α, phorbol 12-myristate 13-acetate (PMA), valporic acid and combinations thereof.
9. The cell line of claim 1 , wherein the provirus contains an intact HIV-I gag gene, thereby allowing the use of Gag expression as a further marker of viral gene expression.
10. The cell line of claim 1 , wherein the latent provirus in the cell line is replication-incompetent.