Testing Compounds for Antiviral Activity in Cell Cultures Infected with HIV

This chapter describes the procedures that can be used to determine compounds that have antiviral activity against HIV. These include: maintenance of lymphoblastoid cell lines, preparation of peripheral blood mononuclear cells (PMMCs), and determination of the infectivity of the HIV stock-supernatant and antiviral assays. The assays described use both acutely and chronically infected cells. Toxicity of compounds is assessed by measuring ¹⁴ C uptake. These protocols are used for the evaluation of compounds that can be carried out by a single individual in a Category 3 containment laboratory. The number of compounds analyzed would be about 10, which is a convenient number to fill a single 96-well p24 enzyme-linked immunosorbent assay (ELISA) plate.