Acute infection of most primary cells and cell lines with HIV depends upon the sequential engagement of CD4 (primary receptor) and a chemokine coreceptor (usually CCR5 or CXCR4) by gp120 Env. Chronically infected cell lines and clones are currently used as sources of virus for infecting other cell types, as "factories" for large-scale production of virions or viral components, and as model systems for studies of regulation of virus expression. This unit provides detailed protocols for acute in vitro HIV infection of primary T cell blasts, interleukin-2-stimulated PBMC, and resting PBMC. The unit also contains information on how to determine the chemokine coreceptor usage of the virus for experimental infections. The use of cell lines as targets of acute infection is also described. Finally, protocols for generating and studying chronically HIV-infected cell lines are provided.